Size characteristics of asbestiform fibers in lung tissue of animals that grazed in areas rich in ophiolitic outcrops in Central Calabria (Southern Italy).

In Calabria (Southern Italy) naturally occurring asbestos (NOA) mainly occurs in the ophiolitic sequences cropping in the Mount Reventino area. The most common type of asbestos detected was the amphibole tremolite; fibrous antigorite and minor chrysotile were also found. The development of asbestos-related diseases depends on, among other things, the morphological characteristics of fibers, length and width, affecting the durability of asbestos fibers in the lung. In this work fifteen lung samples of sheep, goats and wild boars, grazing around the Mount Reventino area were collected and asbestos fibers analysed. Observed fibers (357), of which 97 % were tremolite and 3 % antigorite fibers, were grouped according to species, grazing area and age of the animals. The aim of this work was to highlight any differences among the groupings and to compare our size results with data in literature related to exposed populations. Principal Component Analysis (PCA) highlighted a positive correlation between tremolite fiber length and width and revealed groupings in terms of animal age. The Kruskal-Wallis test showed statistically significant differences between fiber mean widths in young and old animals. 63 % observed asbestiform fibers were longer than 5 µm and 7 % of the fibers were longer than 20 µm (critical fiber length connected to the frustrated phagocytosis by the macrophage). Fibers conforming to the Stanton Hypothesis size (predictor of the carcinogenic potency of fibers) were 1 %. Our size parameters of fibers detected in the animal lungs were in fairly good agreement with literature data for human asbestos exposure to tremolite. These results confirmed that an animal-sentinel system could be used to monitor the natural background of the airborne breathable fibers exposure. In addition, the size correlation of animal-human breathed fibers could be useful to study their potential toxicity. Additional data are necessary for improving the agreement with human exposure data.

Round-Robin Tests with Relocatable Coverslips Improve Asbestos Fibre Counting.

Asbestos fibre counting by phase-contrast microscope is subject to many sources of variation, including those dependent on the analyst. In this study, asbestos sample slides prepared with relocatable coverslips have been used for fibre counting among voluntary analysts to evaluate their proficiency. One slide of amosite and one of chrysotile were distributed to all the analysts, and three proficiency testing rounds were conducted for amosite and four for chrysotile. Each relocatable coverslip has a report in which are reported for each viewing field both the number of certified fibres (Verified Fibres) and a drawing representing the shape and position of the individual fibres. In the first round, the analysts were asked to report only the number of fibres counted in each of the predesignated fields of view. In the other rounds, subsequently developed, the analysts had to report the number and the position of the fibres for each field. The reported number of fibres and their position in each of the designed fields were evaluated against their respective verified fibres, to identify types of error. Discrepancies between reported fibres and verified fibres in each field of view have been used to evaluate the proficiency of the analysts. The discrepancies can be positive (D+) or negative (D-) depending on whether the analyst counts, for a specific field of view, more or less fibres compared to the verified fibres. The score is calculated using the following equation: Score = (1 - ∑D+ + ∑│D-│/VF) × 100. An analyst obtaining a score of ≥60, which corresponds to (∑D+ + ∑│D-│)/VF ≤ 0.40, is proficient. The number of laboratories that participating in this study varied from 13 to 17 depending on the rounds. For amosite fibre counts, the results were generally good compared to a proficiency score of 60. The major error made by analysts was the counting of fibres shorter than 5 µm, where this error was of 62% of extra fibres and accounted for 8% over-estimation of amosite fibres. For chrysotile, a score of ≥50 has been used to consider an analyst as proficient. The results of chrysotile fibres showed that in the first round all analysts counted less than fifty per cent of the verified fibres. In the second round 10 analysts out of 13 reached a score of ≥50, 8 of 16 in the third and 10 of 12 in the fourth. For chrysotile fibres, the error relating to the counting of fibres shorter than 5 µm was of 56% of extra fibres, but the error that most influenced the results was the number of oversight-missing fibres. This type of error accounted for 97% of the missing fibres and for the 29% under-estimation of the chrysotile fibres. For amosite fibre counting, results of this study show an improvement of the analyst's performance. For the chrysotile fibre count, although there is a significant improvement in the comparison between some rounds, this is not continuous over time.

Asbestos containing materials in schools of Rome and surrounding area (Italy).

Despite Italy banning use and production of asbestos in 1992, it continues to represent a risk to human health due to its permanence in the places where it was located. The aim of this work is to estimate how many schools in Rome (Italy) have asbestos containing materials (ACM), and to assess whether the location, condition and nature of ACM can influence the level of risk for student health. 3,672 schools were contacted and 1,451 participated to asbestos survey. 692 bulk samples were collected and analyzed by optical and electron microscope and Fourier transform infrared spectrometer. About 16% of the surveyed schools had ACM. Most of the ACM were not accessible to students (water tanks, boiler thermal insulations). Asbestos-cement materials and vinyl floor tiles were the most common non-friable materials found in schools and equipment insulation linings and Bunsen burner gauze mats were the friable ACM found in science laboratories. Measures to prevent or reduce asbestos hazards were applied where necessary. This study allowed to develop the awareness of the asbestos health hazard among headteachers and the need to manage these hazards appropriately. It represents the beginning of a larger study leading to ACM national mapping in schools.

Primary Ovarian Mesothelioma: A Case Series with Electron Microscopy Examination and Review of the Literature.

Primary ovarian mesothelioma is a rare, aggressive neoplastic disease with a poor prognosis. At onset, the tumor is only rarely limited to the ovaries and usually already widespread in the peritoneum. The rarity of this entity and the difficulties differentiating it from either ovarian carcinoma or peritoneal mesothelioma may lead to frequent misdiagnoses and may raise some concerns about its histogenesis. Thus, reporting such rare cases is fundamental to gain greater awareness of this neoplasm and try to answer unsolved questions. Herein, we described four cases of histological diagnoses of ovarian mesothelioma extrapolated by the regional mesothelioma register of Apulia (southern Italy). In all cases, a detailed medical history was collected according to national mesothelioma register guidelines. A broad panel of antibodies was used for immunohistochemistry to confirm the diagnoses. Moreover, ovarian tissue samples were also examined by transmission and scanning electron microscopy, detecting asbestos fibers and talc crystals in two cases. Because of the few cases described, we reviewed the English literature in the Medline database, focusing on articles about ovarian mesothelioma "misclassification", "misdiagnosis", "diagnostic challenge" or "diagnostic pitfall" and on unsolved questions about its histogenesis and possible risk factors.

Environmental contamination by naturally occurring asbestos (NOA): Analysis of sentinel animal lung tissue.

Ophiolites are known sources of naturally occurring asbestos (NOA). In Calabria (Southern Italy) NOA are mainly concentrated in the ophiolitic sequences cropping in the Mount Reventino area, in the southern part of the Sila massif, and along the Coastal Chain. The most common type of asbestos identified in the rocks of these areas belongs to the tremolite-actinolite series. Another identified asbestiform mineral is fibrous antigorite belonging to the serpentine mineral group with a minor amount of chrysotile. The purpose of the present study is to evaluate the diffusion of natural asbestiform fibers from NOA using sentinel animals. Fifteen lung samples of sheep, goats and wild boars from Mount Reventino area and two from an area free from NOA were collected. The lung samples were subjected to anatomopathological examination and lung fiber burden analysis by electron microscopy. Abundant tremolite and few antigorite fibers were detected in the lung samples coming from the NOA area. No corpuscle of asbestos was observed. No fiber was found in the two lung samples of sheet from the area free from NOA. These concentrations of fibers per gram of dry weight of lung tissue (f/gdw) ranged from 104 to 106 f/gdw. The asbestos fibers detected in the lungs of the examined animals reflect the geological features of the areas where they grazed and lived. The anatomopathological analysis showed that 60% of the examined animals had macroscopic lesions affecting their lungs. The presence of tremolite fibers in the lungs confirms the diffusion of mineral fibers in the environment and the real advantage of using animal populations in the study areas.

[Health impact of asbestos fibres naturally occurring in Mount Pollino area (Basilicata Region, Southern Italy)]. / Impatto sulla salute delle fibre di amianto presenti in affioramenti naturali nell'area del Pollino (Basilicata).

OBJECTIVES: to estimate the health impact of asbestos fibres naturally occurring in Mount Pollino area (Basilicata Region, Southern Italy). DESIGN: geographic mortality, hospitalization, and incidence study. Setting and participant s: population resident in 12 Municipalities of Mount Pollino area with naturally occurring asbestos fibres. MAIN OUTCOME MEASURES: standardized mortality ratio (SMR) and standardized hospitalization rate (SHR) for asbestos-related diseases; standardized incidence ratio (SIR) for mesotheliomas. Result s: in the area of Mount Pollino, where asbestos fibres naturally occur, especially in the sub-area in which fibres are close to dwellings and settlements, it was observed: • a significant excess of mesothelioma incidence (SIR: 208; CI95% 111-355; 13 observed); • a non-significant excess of hospitalization for malignant pleural neoplasms (SHR: 176; CI95% 93-335; 9 observed); • a significant excess for mortality and hospitalization for pneumoconiosis (SMR: 534; CI95% 345-824; 20 observed - SHR: 245; CI95% 149-405; 15 observed); • a significant excess for hospitalization (SHR: 852; CI95% 290-2,506; 3 observed) for asbestosis. CONCLUSION: it is necessary to continue environmental monitoring and environmental remediation in the area with higher asbestos exposure. It is suggested to implement a permanent process of epidemiological surveillance in this same area. A communication plan with local administrators, general practitioners, school teachers, media, and the resident population at large should be realized.

Prevention of disease caused by fluoro-edenite fibrous amphibole: the way forward.

Few months after the publication of the monographic section of Annali dell'Istituto Superiore di Sanità second issue of 2014 "Health impact of fibres with fluoro-edenitic composition", the carcinogenicity of fluoro-edenite was assessed by the International Agency for Research on Cancer (IARC) in the frame of Monograph 111. The IARC Working Group concluded that there is sufficient evidence in humans that exposure to fluoro-edenite fibrous amphibole causes mesothelioma, and sufficient evidence of carcinogenicity in experimental animals. Fluoro-edenite was allocated to Group 1 (the agent is carcinogenic to humans). Now, in view of the recent IARC evaluation, preventive action in Biancavilla requires an upgrade. First of all, environmental monitoring has to be further implemented. All operations of house cleaning should be performed employing wet tools, in order to avoid dust-raising. It is very important that environmental and biological monitoring be related to epidemiological surveillance. The recently approved act of the Sicilian Government concerning a plan of health interventions in Biancavilla will favour cooperation between national, regional and local health institutions with the common goal of improving the quality and appropriateness of diagnostic and therapeutics procedures offered by the health services.

Environmental concentrations of fibers with fluoro-edenitic composition and population exposure in Biancavilla (Sicily, Italy).

INTRODUCTION: The town of Biancavilla (Sicily) was included in the National Priorities List of Contaminated Sites due to environmental dispersion of amphibole fibers owing to the extraction of materials from a local quarry. The present report summarizes results from several, hitherto unpublished, environmental surveys carried out in the area, as well as from published analyses of the chemistry and composition of fibers. METHODS: Data included here comprises environmental fiber concentrations by scanning electron microscopy with energy dispersive spectroscopy (SEM-EDS) analysis in soil, indoor and outdoor air, personal monitoring, as well as a chemical characterization of the fibers. The full chemical structure and spectroscopic characterization of fibers were obtained through a multi-analytical approach: SEM-EDS, X-ray powder diffraction (XRPD), as well as Mössbauer (MS) and Fourier transform infrared (FT-IR) spectroscopies. RESULTS: Data analyzed provided a spatial and temporal picture of fiber concentrations in Biancavilla, and a qualitative assessment of population exposure. Results suggest that until 2000, the population had been exposed to high levels of amphibole fibers. Mitigation measures adopted since 2001, gradually reduced exposure levels to about 0.1-0.4 ff/l. Previous studies on fibrous amphiboles from Biancavilla reported considerable chemical variability. Differences in composition, especially concerning the presence of Si, Ca, Fe, and Na, were found both within and between samples. Compared to the previously investigated prismatic fluoro-edenite, these fibrous fluorine amphiboles consistently showed higher average values of Si and Fe content, whereas Ca was significantly lower, which we consider a distinctive characteristic of the fluorine fibrous variety. CONCLUSIONS: The population of Biancavilla had been highly exposed to a suite of fibrous amphiboles for over 50 years. Dust mitigation measures have gradually reduced exposure, but continuous environmental follow-up is necessary in order to monitor exposure levels and prevent adverse health effects for future generations.

Quantitative energy dispersive X-ray analysis of submicrometric particles using a scanning electron microscope.

The quantitative scanning electron microscope-energy dispersive X-ray (SEM-EDX) analysis of a horneblende and two augite prismatic samples reduced to submicrometric particles was performed, and error due to the particle effects ("absent mass" and the "reduced absorption" effect) was minimized. Correction factors as a function of fragment size were obtained for O, Na, Mg, Si, Ca, and Fe. In addition, the influence of chemical composition of the samples used as standards (the matrix effect) on correction factors was evaluated. The results indicate that the absent mass effect is dominant for all elements except for the light elements O and Na, for which the reduced absorption effect is dominant. No significant matrix effect has been observed. By using corrected SEM-EDX data, the error on quantification of the element concentration has been estimated to be 3% relative for light elements and below 2% relative for heavy elements (notably, about 1% relative for Fe).

[Pleural mesothelioma and environmental exposure to mineral fibres: the case of a rural area in the Basilicata region, Italy]. / Mesotelioma pleurico ed esposizione ambientale a fibre minerali: il caso di un'area rurale in Basilicata.

The main aspects of the sites characterized by environmental exposure to mineral asbestiform fibres are described. Several adverse health effects including high incidence of pleural mesothelioma are reported. The average concentration of airborne fibres is generally low but it rises significantly in association with mechanical disturb of materials with fibres. Multiple sources of exposure have been identified, fibres can be found in the soil and in many materials locally used, mainly in buildings. Three mesothelioma cases were observed in a small rural area of the Basilicata region (Italy). Two of them had a possible occupational exposure to asbestos, the third had a proved environmental exposure to tremolite. This fibre, found in the area, is the same observed in two of the three biological samples analysed.

Pulmonary fibre burden in sheep living in the Biancavilla area (Sicily): preliminary results.

In a national survey on mortality from malignant pleural neoplasms in Italy, aimed at detecting geographic clusters of cases of the disease, the town of Biancavilla, located in a volcanic area of Eastern Sicily, showed high risk of pleural mesothelioma in the absence of occupational asbestos exposure. An environmental survey suggested the stone quarries located in 'Monte Calvario', south-east of the town of Biancavilla, as a possible source of asbestiform fibre exposure. A subsequent crystal-chemistry investigation of the 'Monte Calvario' amphiboles identified the mineral asbestiform fibres as 'fluoro-edenite', a new end-member of the edenite ==> fluoro-edenite series. A collaborative epidemiological and environmental study was initiated to investigate the characteristics of the outbreak of malignant mesothelioma and test the hypothesis of a causal association with exposure to naturally occurring fibres. To investigate if a sheep population could be used to monitor the environmental diffusion of the fibres, we examined lung specimens from 27 culled sheep, at least 3 years old and living near Monte Calvario to check for the presence of fluoro-edenite fibres, using scanning electron microscopy (SEM) analysis and X-ray microanalysis. Fourteen mineral species have been isolated in the mineral particulate matter taken from pulmonary parenchyma, and fluoro-edenite was detected in eight sheep. These preliminary data suggest a possible bio-indicative role of sheep as sentinel animals in the evaluation of environmental fibre diffusion, which merits further investigation.

Effects of the new-identified amphibole fluoro-edenite in lung epithelial cells.

An epidemiological survey on mortality for malignant pleural neoplasm in Italy evidenced a number of patients in Biancavilla, a village located in a volcanic area of eastern Sicily, none of which had been significantly exposed to asbestos during their professional lives. Environmental studies suggested the involvement of the material derived from stone quarries in the disease onset. A detailed crystal-chemical analysis of amphiboles contained in this material allowed the discovery and the identification of a new fiber that was named fluoro-edenite. In order to define the mode of action of fluoro-edenite at a subcellular level, we have conducted a study by using A549 cells, a tumor-cell line from a human lung carcinoma with properties of alveolar epithelial cells. The results obtained showed a remarkable tropism of A549 cells toward fluoro-edenite fibers. In fact, these epithelial cells contacted the fibers via the extension of membrane ruffles and filopodia that allowed the capture and most probably the internalization of material into the cytoplasm. Moreover, fluoro-edenite interfered with epithelial cell physiology, by reducing the proliferation rate without perturbing the cell cycle and increasing the release of the proinflammatory cytokine IL-6, one of the main mediators of asbestos-induced pathophysiological response.