European Council of Legal Medicine (ECLM) on-site inspection forms for forensic pathology, anthropology, odontology, genetics, entomology and toxicology for forensic and medico-legal scene and corpse investigation: the Parma form.

Further to a previous publication by the European Council of Legal Medicine (ECLM) concerning on-site forensic and medico-legal scene and corpse investigation, this publication provides guidance for forensic medical specialists, pathologists and, where present, coroners' activity at a scene of death inspection and to harmonize the procedures for a correct search, detection, collection, sampling and storage of all elements which may be useful as evidence, and ensure documentation of all these steps. This ECLM's inspection form provides a checklist to be used on-site for the investigation of a corpse present at a crime or suspicious death scene. It permits the collection of all relevant data not only for the pathologist, but also for forensic anthropologists, odontologists, geneticists, entomologists and toxicologists, thus supporting a collaborative work approach. Detailed instructions for the completion of forms are provided.

Evaluating the presence of SARS-CoV-2 RNA in the particulate matters during the peak of COVID-19 in Padua, northern Italy.

The airborne transmission of SARS-CoV-2, the etiologic agent of the current COVID-19 pandemic, has been hypothesized as one of the primary routes of transmission. Current data suggest a low probability of airborne transmission of the virus in open environments and a higher probability in closed ones, particularly in hospitals or quarantine facilities. However, the potential diffusion of the virus in open environments, especially using particulate matter (PM) as a transport carrier, generated concern in the exposed populations. Several authors found a correlation between the exceeding of the PM10 concentration limits in some Italian cities and the prevalence of Covid-19 cases detected in those areas. This study investigated the potential presence of SARS-COV-2 RNA on a representative series of PM samples collected in the province of Padua in Northeastern Italy during the first wave of COVID pandemic. Forty-four samples of PM2.5 and PM10 were collected between February 24 and March 9, 2020 and analyzed with RT-qPCR for SARS-CoV-2 RNA. The experimental results did not indicate the presence of SARS-CoV-2 RNA in the outdoor PMs, thus confirming the low probability of virus airborne transmission through PM.

COVID-19 pandemic: an Italian single institution's experience and lessons learned by public health residents' workforce.

Methods: We hereby provide a systematic description of the response actions in which the public health residents' workforce was pivotal, in a large tertiary hospital. Background: The Coronavirus Disease 2019 pandemic has posed incredible challenges to healthcare workers worldwide. The residents have been affected by an almost complete upheaval of the previous setting of activities, with a near total focus on service during the peak of the emergency. In our Institution, residents in public health were extensively involved in leading activities in the management of Coronavirus Disease 2019 pandemic. Results: The key role played by residents in the response to Coronavirus Disease 2019 pandemic is highlighted by the diversity of contributions provided, from cooperation in the rearrangement of hospital paths for continuity of care, to establishing and running new services to support healthcare professionals. Overall, they constituted a workforce that turned essential in governing efficiently such a complex scenario. Conclusions: Despite the difficulties posed by the contingency and the sacrifice of many training activities, Coronavirus Disease 2019 pandemic turned out to be a unique opportunity of learning and measuring one's capabilities and limits in a context of absolute novelty and uncertainty.

Crime scene and body alterations caused by arthropods: implications in death investigation.

The activity of arthropods on corpses has been largely investigated, since they can produce information to reconstruct the peri-mortem events. However, the feeding/movement activity of insects around the crime scene, among the clothes and on the body, can also cause some alterations that can lead to wrong reconstruction and misinterpretations. This article summarises all the post-mortem arthropods artefacts related to the scene (i.e. fly artefacts and floor stripes) and the body (i.e. skin and other soft tissue alterations, bone alterations and hair alterations) that can mislead the forensic pathologist, discussing macroscopic and microscopic findings derived from forensic casework and from experimental laboratory studies, in order to provide a useful instrument to avoid misinterpretations and evaluation errors. Finally, some procedural notes for the documentation and the interpretation of findings are proposed.

Entomotoxicology in burnt bodies: a case of maternal filicide-suicide by fire.

One of the most common methods of maternal filicide is by fire. In this case study, a 40-year-old female and her children were found completely burned in a burnt out car. All bodies showed a degree of destruction by fire consisting to a level 3 of the Crow-Glassman Scale (CGS) and early stage of insect activity. Toxicological analyses were performed on soft tissues and body fluids still available. The results were positive for diazepam and its metabolites only for children with blood concentrations consistent with therapeutic doses of benzodiazepines. Home video surveillance cameras confirmed sedation prior to death recording the mother while administering some drops of sedative drugs in a soft drink to the children just a couple of hours before setting fire to the car. Based on autopsy findings, all victims were still alive at the time of fire. The cause of death was determined as carbon monoxide poisoning and fatal thermal injuries by fire. This case study has a special focus on the entomotoxicology and the potential role of insects in death investigations of burnt bodies, supposed to be an inadequate substratum for insect colonization. It demonstrates that in burnt bodies, arthropod colonization can be quite immediate after fire is extinguished. Toxicological analyses performed on larvae actively feeding on the children's bodies were positive for diazepam and its metabolites in small amount compared with blood concentrations, whereas the larvae collected from the mother's body were totally negative. These data, according to the autopsy findings and the toxicological results from the victim's blood and tissues, supported the suspect of a non-lethal sedation prior to death, which is a common behaviour in maternal filicide.

Multiple deaths caused by a fire in a factory: identification and investigative issues.

The article presents a case of multiple casualties following a textile factory fire. The incident required a full DVI team similar to large mass-disaster because of the specific operational aspects and identification difficulties. The autopsy results were consistent with death by fire and the toxicological investigations revealed carbon monoxide poisoning in four cases (HbCO% ranging between 88,05 and 95,77), two deaths by cyanide intoxication (with concentrations between 5,17 and 8,85 mcg/ml), and in one case there was a synergistic effect of the two substances (carbon monoxide and cyanide). The identification, carried out in accordance with INTERPOL protocols, encountered serious difficulties in the AM phase primarily due to a language barrier and the lack of any dental or medical information relating to the victims. Secondary identifiers proved to be very useful in corroborating possible identities. As a result of the combined efforts of a team of experts the identity of each victim was determined and in all cases at least one primary identifier was used in the identification process. The deployment of DVI teams composed of forensic experts from different fields of expertise and well acquainted with DVI procedures, is essential in events involving multiple casualties that may also include foreign victims. The DVI team should intervene not only in PM examinations but also in the collection of AM data for those individuals not accounted for and by helping police in contacting families of missing people.

Myiasis by Megaselia scalaris (Diptera: Phoridae) in a python affected by pulmonitis.

Myiases are caused by the presence of maggots in vertebrate tissues and organs. Myiases have been studied widely in humans, farm animals, and pets, whereas reports of myiasis in reptiles are scarce. We describe a case of myiasis caused by the Megaselia scalaris (Loew) in an Indian python (Python molurus bivittatus, Kuhl) (Ophida: Boidae). The python, 15 yr old, born and reared in a terrarium in the mainland of Venice (Italy), was affected by diffuse, purulent pneumonia caused by Burkholderia cepacia. The severe infestation of maggots found in the lungs during an autopsy indicated at a myiasis.

First evaluation of the serum level of anti-hepatitis B surface antigen after vaccination in Libya.

The hepatitis B virus (HBV) vaccination schedule in Libya follows international recommendations (1st dose at birth, 2nd after 1 month and 3rd after 6 months). This research aimed to evaluate the long-term protection of the HBV immunization programme in Tripoli and to determine the best age to administer booster doses. Serum levels of hepatitis B surface antigen were determined in 277 randomly selected children aged 1-12 years. The response to HBV vaccine in 1-3-year-olds was 93.2%, but this declined with age and at 7-9 years after initial vaccination only 53.1% of children had protective titres (> or = 10 mIU/mL). No significant differences between males and females in antibody persistence or response to vaccine were observed. We recommend continuing the HBV vaccination programme and that a booster dose be given to 6-year-old children to ensure maximum protection during the period of school entry and beyond.

Nosocomial myiasis in a patient with diabetes.

This report describes a case of nosocomial myiasis caused by larvae of Sarcophaga (Bercaea) africa. The parasitosis developed in an ulcer on the heel of a patient with type 2 diabetes. The ulcer was dressed when the deposition occurred. An experiment was performed in order to demonstrate the ability of Sarcophaga larvae to move through bandages and reach purulent wounds. This report highlights the need for particular attention, education and specific protocols in hospitals in order to avoid myiasis, which can compromise the organization's reputation for hygienic standards with possible legal consequences.

Insects found on a human cadaver in central Italy including the blowfly Calliphora loewi (Diptera, Calliphoridae), a new species of forensic interest.

In the case of unidentified bodies the estimation of the period since death or of the season of death plays an important role to focus the attention on a reduced number of people among the ones reported missing. Forensic entomology can be one of the most important methods for these estimations, as occurred in this case. Flies are typically the first insects to colonize a dead body. The case reported here concerns the colonisation by insects of a male body in advanced decay found during the winter in Central Italy. This case is of particular interest as few data are available on the entomological evidence in the cold season. In particular, in this case we recovered Calliphora loewi (Calliphoridae), a species never collected before on dead bodies in Southern Europe. Larvae of the black soldier fly Hermetia illucens (Stratiomyidae), pupae and larvae belonging to genus Hydrothea (Muscidae), and Necrobia rufipes (Cleridae) specimens were also collected. The estimated PMI enabled identification of the cadaver, confirmed by DNA analysis.

Records of Chrysomya albiceps in Northern Italy: an ecological and forensic perspective.

Knowledge of the carrion-breeding insects present at a local level is important and necessary for defining the post-mortem interval. Climate changes and globalisation are affecting species ranges and population dynamics. In this note, we report the incidence of Chrysomya albiceps (Diptera: Calliphoridae) on dead human bodies and carrion in Northern Italy. These data confirm the spread of this species in the Northern regions. The partial sequencing of a 583-bp region of the cytochrome oxidase subunit 1 gene of an Adriatic population did not reveal any difference compared to the same genomic region in the African and South American populations of this species.

Records of Chrysomya albiceps in Northern Italy: an ecological and forensic perspective

Knowledge of the carrion-breeding insects present at a local level is important and necessary for defining the post-mortem interval. Climate changes and globalisation are affecting species ranges and population dynamics. In this note, we report the incidence of Chrysomya albiceps (Diptera: Calliphoridae) on dead human bodies and carrion in Northern Italy. These data confirm the spread of this species in the Northern regions. The partial sequencing of a 583-bp region of the cytochrome oxidase subunit 1 gene of an Adriatic population did not reveal any difference compared to the same genomic region in the African and South American populations of this species.

Use of Lucilia species for forensic investigations in Southern Europe.

The aim of this study was to highlight the importance of evaluating entomological evidence in forensic investigations on a regional scale. To evaluate climatic, geographical and environmental influences on the selection of carrion-breeding fauna in Northern Italy and consequently on inferred forensic data (post-mortem intervals and post-mortem transfer), we present details of six indoor-outdoor cases. Results show that the most abundant species was Lucilia sericata, together with other fly species of entomo-forensic interest, belonging to the Calliphoridae and Sarcophagidae families. In particular, for the first time in Italy, we report finding Phormia regina, Lucilia ampullacea, Lucilia caesar and Sarcophaga (Pandelleana) protuberans on fresh cadavers. The active period of L. sericata in Northern Italy, according to previous findings in Southern Europe, revealing clearcut differences with phenologies in Northern Europe, has important consequences in estimating the period (season, months) of death in cases of long post-mortem intervals (several months or years) if empty puparia of this fly are found. According to our results, the distribution of L. sericata in areas with urban sprawl, like Northern Italian regions, cannot be used to evaluate post-mortem transfer from an urban area to a rural one.

[Forensic entomology and globalisation]. / L'Entomologia forense e la globalizzazione.

The main aim of Forensic Entomology has always been, and is today, to establish the time of death (P.M.I.: Postmortem Period) or, more exactly, how long a carrion has been exposed in the environment. Most of the invertebrate fauna occurring on corpses consists of insects (mostly Diptera and Coleoptera). They are selectively attracted by the decomposing status of the carrion, and form complex communities or biocenosis within necrophagous or sarcophagous species and their predators, parasites and parasitoids, competing each one another. The rapid and continuos changes of the micro-ecosystem (the body), until its breakdown, does not permit the achievement of a steady state or an equilibrium in the animal communities. These continuous modifications give us the possibility to estimate when (and where) the death has occurred, by the identification of the species feeding on the corpse, the knowledge of their life history, and the length of each stage of their cycle at varying the temperature and the other abiotic factors, external to the carrion ecosystem. The P.M.I. today is still largely based on the tables of faunal succession on human cadavers recognised by Mégin in 1894, with few changes proposed by Authors from other countries. In the last years, however, it happens more and more often, that the natural communities are subverted by the presence of allocton species, which can compete, predate or parasite the most common local sarcophagous insects, modifying, this way, the succession waves and the trophic nets. The immission in the environment of foreign species may be voluntary or casual, but in any case is due to anthropic activities. The voluntary immission happens when some species, employed in the biological struggle against pest or dangerous insects, for pollination of allocton plants, or for other commercial utilities, are beyond man's control and swarm onto the environment; the casual spread is due to the globalisation phenomenon, that distributes the "little organisms" by chance, together with travellers, goods and food items. Together with human migratory flows, raw materials and vegetal foodstuffs travelling from Tropical developing Countries to the North of the world, also many tropical and subtropical insects can be carried out from their original lands. Eurioecious and polyphagous species, and species that evolved mechanisms (as diapausa or hibernation) to get over critical environmental conditions have the highest probability of survival. Saprophagous insects, and flies in particular, evolved such capabilities. The mortality of foreign species due to the difference of temperature and seasonally between tropical and temperate areas was, in the past, the most effective factor limiting the geographic propagation of insects. The ongoing global climate changes induce insect populations, now confined to the tropics, to most likely spread towards middle latitudes, where their specific competitors, predators and parasites, which regulate the population growth, are often absent. The lack of a biological control, the warming up of atmosphere temperature and the fall of the differences among seasons induce a more rapid development and an increased number of generations in new species, that often displace the autocton ones. This phenomenon is much more clear in little and simple ecosystems (such as carrion), mostly occur than in large and complex ecosystems, where many more components, vegetal organisms and phytophagous animals included, are present. To demonstrate how globalisation and climate changes are breaking the geographic barriers, we present some cases in which, during our entomoforensic investigations, performed mainly in North-eastern Italy, Neotropical, African and Asiatic necrophagous flies, beetles and wasp parasitoids have been collected, some of which rare or new for Italy or Paleartic Region. In particular, we report our studies on the american black soldier fly, Hermetia illucens (Diptera: Stratiomyidae), that is showing a heavy competition with the local saprophagous species and is reaching great importance in some man activities and in cattle health.

Capillary zone electrophoresis with optimized temperature control for studying thermal denaturation of proteins at various pH.

Capillary electrophoresis (CE) was used to analyze the thermal denaturation of bovine beta-lactoglobulin at different pH. This model protein exhibits complex pH- and temperature association/dissociation dependence balances in its quaternary structure. The study was possible after modification and improvement of a capillary electrophoresis apparatus. The improvement allowed both efficient control (temperature fluctuations <0.05 degrees C) and accurate measurement of the temperature (+/- 0.1 degrees C) within the capillary cartridge. CE allowed the thermodynamic parameters of beta-lactoglobulin thermal denaturation to be estimated. The transition temperature, Tm, was determined at acidic, neutral and alkaline pH. Van't Hoff analysis was performed through direct measurement of native and unfolded protein populations in the slow-time regime. This allowed estimation of thermodynamic parameters (deltaH, deltaS, deltaCp). Finally, the stability curve, i.e., the temperature dependence of the free energy change (deltaG) of protein unfolding was drawn. The accuracy of the parameters values compares with parameters obtained by calorimetric measurements. The available parameters and the requirement of minute amount of protein sample are of potential interest in the field of protein engineering and biological pharmaceuticals. Accordingly, CE can be proposed as a convenient tool to study protein stability and denaturation processes.

Blocking the cycle of panic disorder. Ways to gain control of the fear of fear.

Panic disorder is a common and potentially debilitating anxiety disorder that can adversely affect patients' personal, social, work, and academic lives. It may go unrecognized for long periods, because physical symptoms may mimic other conditions. Typical onset is during late adolescence and early adulthood. Stressors (e.g., new environments, loss of social support, relationship difficulties, academic pressure, substance use) may trigger or exacerbate panic symptoms. Evaluation by a healthcare professional paying close attention to medical and psychological aspects of panic disorder can lead to a proper diagnosis and effective management with medication (e.g., SSRIs) and psychotherapy.