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.

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.

Biomonitoring of workers employed in a titanium dioxide production plant: Use of buccal micronucleus cytome assay as noninvasive biomarker to evaluate genotoxic and cytotoxic effects.

We aimed to evaluate whether TiO2 production process induces genotoxic and cytotoxic effects on the first target organ of inhalable particles by a sensitive and noninvasive biomarker of effect. Final aim was to find a useful and suitable tool to assess and manage the risk of TiO2 occupational exposure. We enrolled 40 workers employed in TiO2 production, 5 office workers, and 18 external controls. Buccal micronucleus cytome assay (BMCyt assay) was applied because it allows to evaluate micronucleus (MN), nuclear buds (NB), and broken eggs (BE) indicating the presence of chromosomal instability and gene amplification and binucleated cells (BIN), karyolytic cells (KL), and condensed chromatin (CC) indicating cytokinesis defect or arrest, cell death and apoptosis respectively. We characterized the exposure measuring inhalable and respirable particles by personal monitoring. BMCyt-assay showed in exposed workers compared with external controls a higher value of MN frequency (2.57 vs. 0.05‰, p < .001) and MN positivity, evaluated as percentage of subjects with MN frequency higher than a 1.5‰ cut-off value (52.5 vs. 0%). We also found in exposed workers higher frequency of BE + NB (2.41 vs. 0.22‰, p = .002), BIN (9.45 vs. 8.44‰, p = .047) and CC (1.80 vs. 0.21, p = .001) than in controls. Moreover, we found a relationship between personal monitoring results and presence of MN and other cellular anomalies. This study demonstrates induction of genotoxic and cytotoxic effects on buccal cells of workers involved in TiO2 production, suggesting the suitability of BMCyt assay as tool for risk assessment and management of TiO2 exposure.

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.

Biosolubility of high temperature insulation wools in simulated lung fluids.

OBJECTIVE: Biosolubility is an important parameter in the understanding of mechanisms involved in pulmonary toxicity of fibrous materials. It can be studied in vitro using models of simulated lung fluids and observing the loss of structural molecules, expressed as dissolution constant (Kdis). The aim of this paper was the study of dissolution behaviour of four wools belonging to high temperature insulation wools (HTIW) in saline solutions simulating lung fluids. METHODS: Four HTIW were studied in saline solutions at pH 7.4 (representative of the extracellular environment) and 4.5 (representative of the intracellular conditions): refractory ceramic fibers (RCF), two alkaline earth silicate wools (AES1 and AES2 with high calcium and magnesium content respectively), and polycrystalline wools (PCW). Size, morphological and chemical changes of fibers were observed by scanning electron microscopy (SEM) with energy-dispersive X-ray spectrometry (EDS) and inductively coupled plasma atomic emission spectrometry (ICP-AES). RESULTS: RCF, AES2 and PCW did not show statistically significant diameter changes. AES1 size distribution shifted to a larger mean diameter suggesting that through dissolution there was a preferential loss of thin fibers at acid pH after 14 days of treatment.Both AES wools showed selective leaching of alkali/alkali earth oxides (incongruent dissolution) at pH 7.4: a fast and extensive selective leaching of calcium for AES1 with complete dissolution of fibers already after 14 days of treatment and a moderate selective leaching of magnesium for AES2. PCW showed some transversal breakage of the fibers in both pH environments (low congruent dissolution). For RCF, the treatment produced uncorroded fibers in both pH environments without chemical changes and fiber fragmentation (no dissolution).The estimated Kdis at physiological pH followed the sequence: AES1 > AES2 > PCW > RCF. All wools had a low Kdis at acid pH suggesting a low dissolution rate of short fibers. CONCLUSION: The leaching process and transverse fragmentation play an important role in the biopersistence mechanisms and pathogenicity of fibers and the Kdis estimate is undoubtedly useful as a preliminary toxicological screening of fibers, especially for developing fibers.

Alkaline earth silicate (AES) wools: Evaluation of potential cyto-genotoxic and inflammatory effects on human respiratory cells.

Biosoluble AES wools are increasingly used since considered not hazardous, however, few toxicity studies are available. We evaluated cytotoxic, genotoxic-oxidative and inflammatory effects of two differently soluble AES wools, AES1 (high MgO percentage) and AES2 (high CaO percentage), on alveolar (A549) and bronchial (BEAS-2B) cells. Fiber dimensions and dissolution in cell media were evaluated by SEM analysis. Cell viability, LDH release, direct/oxidative DNA damage (fpg-comet assay) and IL-6, IL-8 and TNF-α release (ELISA), were analysed after 24 h exposure to 2-200 µg/ml. On A549 cells AES1 induced LDH release, slight direct DNA damage and oxidative DNA damage with very high IL-6 release at 100 µg/ml; AES2 induced higher DNA damage than AES1 and slight oxidative DNA damage. On BEAS-2B cells we found direct DNA damage (higher for AES1) and slight oxidative DNA damage (associated to slight increased IL-6 and IL-8 release for AES1). The higher genotoxicity of more soluble AES2 on A549 cells could be explained by higher respirable fibers % and fiber number/µg found after 24 h in RPMI-medium at 100 µg/ml. The higher membrane damage, oxidative DNA damage and inflammation induced by AES1 in A549 cells could be due to the higher DLG and silica percentage. These findings suggest further investigations on AES toxicity.

Asbestiform minerals in ophiolitic rocks of Calabria (southern Italy).

Ophiolitic rocks cropping on Calabria territory, southern Italy, can hold asbestiform minerals potentially harmful for human health. The aim of this work was to detect the fibrous phases of ophiolites along the Coastal Chain of northern Calabria and southern part of the Sila massif. Above 220 massive samples were collected in the study areas and analyzed using optical and electron microscopy, X-ray diffractometry, and Fourier transform infra-red spectrometry. The main fibrous constituent belonged to tremolite-actinolite series followed by fibrous antigorite that becomes more abundant in the samples collected in Reventino Mount surroundings. Results highlighted that serpentinites samples mainly consisted of antigorite and minor chrysotile. Samples collected along the coastal chain of northern Calabria did not hold fibrous materials. The results will be useful for Italian natural occurrences of asbestos (NOA) mapping in order to avoid an unintentional exposition by human activity or weathering processes.

Field comparison of two inhalable samplers used in Italy to measure the wood dust exposure.

BACKGROUND: Particle size affects the performance of personal air samplers used to measure dust exposure in the workplace. Few field studies have been conducted for comparing the performance of personal inhalable samplers. OBJECTIVE: To compare wood dust sampling with two inhalable samplers: IOM (Institute of Occupational Medicine) and Italian cone. METHODS: 136 Italian cone/IOM paired samples and 136 passive IOM samples were collected in 30 Italian woodworking industries. The valid number of sample pairs was 114. Ultra-large particles were collected by passive IOM. The sampling membranes were weighed and the size particles were measured. Mass differences in active and passive IOM samples were calculated (IOM-Δ). RESULTS: Statistical analysis of all 114 Italian cone/active IOM paired results showed a significant mass difference (P < 0.05) and no significant mass difference for sanding and cutting woodworking processes. The Italian cone/IOM-Δ paired results consistently showed no statistically significant mass differences in any woodworking processes. Both samplers performed similarly when ultra-large particles mass contribution was not considered. CONCLUSIONS: These findings confirm the presence of ultra-large particles in woodworking. The Italian cone and IOM samplers can be used interchangeably for personal wood dust exposure assessment, when the wood activities produce small-size particles.

Dissolution of glass wool, rock wool and alkaline earth silicate wool: morphological and chemical changes in fibers.

The behavior of alkaline earth silicate (AES) wool and of other biosoluble wools in saline solution simulating physiological fluids was compared with that of a traditional wool belonging to synthetic vitreous fibers. Morphological and size changes of fibers were studied by scanning electron microscopy (SEM). The elements extracted from fibers were analyzed by inductively coupled plasma atomic emission spectrometry. SEM analysis showed a larger reduction of length-weighted geometric mean fiber diameter at 4.5 pH than at 7.4 pH. At the 7.4 pH, AES wool showed a higher dissolution rate and a dissolution time less than a few days. Their dissolution was highly non-congruent with rapid leaching of calcium. Unlike rock wool, glass wool dissolved more rapidly at physiological pH than at acid pH. Dissolution of AES and biosoluble rock wool is accompanied by a noticeable change in morphology while by no change for glass wool. Biosoluble rock wool developed a leached surface with porous honeycomb structure. SEM analysis showed the dissolution for glass wool is mainly due to breakage transverse of fiber at pH 7.4. AES dissolution constant (Kdis) was the highest at pH 7.4, while at pH 4.5 only biosoluble rockwool 1 showed a higher Kdis.