Laboratory for Process Engineering, Environment, Biotechnology and Energy

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Publications

Occupational Exposure of Firefighters in Non-fire Settings

Publication . Slezakova, Klara; Esteves, Filipa; Vaz, Josiana; Alves, Maria José; Madureira, Joana; Costa, Solange; Fernandes, Adília; Teixeira, João Paulo; Morais, Simone; Pereira, Maria do Carmo

This work assessed firefighters’ exposure to particulate matter (PM) in non-fire work settings during pre-fire season, as a baseline for the respective occupational exposure characterization. Indoor and outdoor air sampling was conducted for two weeks in pre-fire season of 2021 in seven fire corporations (FC1-FC7) in north of Portugal. PM fractions (PM2.5, PM10) were continuously monitored concurrently in indoors (living rooms, rest areas, truck bays) and outdoors. The results showed low levels of pollution. Indoor PM10 was between 2 and 205 μg m–3 (mean 10 μg m–3); PM2.5 were 2—115 μg m–3 (8.5 μg m–3). Both indoor PM fractions were highly and significantly correlated (rs = 0.959–0.997). PM2.5 accounted for 85% of indoor PM; indoor to outdoor ratios (I/O) of PM2.5 ranged between 1.4 and 3.0, thus emphasizing the contribution of indoor emission sources for fine fraction. Outdoor PM10 were 2–6 times higher than indoors (6–894 μg m–3; mean 21 μg m–3); and 2–9 times higher for PM2.5 (5 – 169 μg m–3; 5 μg m–3). Outdoor PM were moderately correlated (rs = 0.584 – 0.878), most likely due to meteorological conditions. Coarse particles contributed the majority of ambient PM10 and accounted for 79%.

2022-09-17Book part

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Development of a Novel Peptide Nucleic Acid Probe for the Detection of Legionella spp. in Water Samples

Publication . Nácher-Vázquez, Montserrat; Barbosa, Ana; Armelim, Inês; Azevedo, Andreia Sofia; Almeida, Gonçalo Nieto; Pizarro, Cristina; Azevedo, Nuno Filipe; Almeida, Carina; Cerqueira, Laura

Legionella are opportunistic intracellular pathogens that are found throughout the environment. The Legionella contamination of water systems represents a serious social problem that can lead to severe diseases, which can manifest as both Pontiac fever and Legionnaires' disease (LD) infections. Fluorescence in situ hybridization using nucleic acid mimic probes (NAM-FISH) is a powerful and versatile technique for bacterial detection. By optimizing a peptide nucleic acid (PNA) sequence based on fluorescently selective binding to specific bacterial rRNA sequences, we established a new PNA-FISH method that has been successfully designed for the specific detection of the genus Legionella. The LEG22 PNA probe has shown great theoretical performance, presenting 99.9% specificity and 96.9% sensitivity. We also demonstrated that the PNA-FISH approach presents a good signal-to-noise ratio when applied in artificially contaminated water samples directly on filtration membranes or after cells elution. For water samples with higher turbidity (from cooling tower water systems), there is still the need for further method optimization in order to detect cellular contents and to overcome interferents' autofluorescence, which hinders probe signal visualization. Nevertheless, this work shows that the PNA-FISH approach could be a promising alternative for the rapid (3-4 h) and accurate detection of Legionella.

2022-07-13Journal article

Open access

Fighting occupational risks among Portuguese wildland firefighters: looking at cytogenetic effects

Publication . Esteves, Filipa; Slezakova, Klara; Madureira, Joana; Vaz, Josiana; Fernandes, Adília; Pereira, Maria do Carmo; Morais, Simone; Teixeira, João Paulo; Costa, Solange

Background:Evidence linking wildland firefighters’ occupational exposure and health outcomes is still limited. Cytogenetic endpoints have long been applied in the surveillance of human genotoxic exposures and early effects of genotoxic carcinogens. Therefore, it is of utmost importance to clarify the exposure-induced cytogenetic effects concerning wildland firefighters’ occupational exposure at different time points (Pre-fire season and fire season). Objective:Here, we aim to evaluate the cytogenetic levels in buccal cells among a group of wildland firefighters during a Pre-fire season, considering both the i) influence of self-reported variables (e.g., lifestyle) on buccal micronucleus cytome assay (BMCyt) outcomes and ii) the cytogenetic damage in exfoliated buccal cells considering the estimated inhalation doses to particulate matter (PM) in non-fire work settings. Methods:A total of 176 northern Portuguese wildland firefighters (82% males; mean age of 37.5 ± 10.9) were recruited during the pre-fire season of 2021. Relevant information was obtained through a self-administered questionnaire. Genomic instability was assessed for 172 northern Portuguese wildland firefighters by BMCyt. PM10 and PM2.5 inhalation doses (indoor/outdoor) were estimated for a group of 80 firefighters based on methods described elsewhere [1]. Results:Some lifestyle variables (e.g., daily consumption of vegetables) shown to have a protective role on some BMCyt endpoints (p<0.05), whereas others such coffee consumption or being partof Permanent Intervention Teams (full-time firefighters) presented a negative impact (p<0.05). No significant association was found between estimated inhaled doses of PM10and PM2.5 (mean 1.73 ± 0.43 μg kg-1and 0.53 ± 0.21 μg kg-1, correspondingly) and BMCyt endpoints. Conclusions:The characterization of a population is a very important step to have a broad perspective of the potential risk factors that may influence the studied endpoints in further analysis. Surveillance based on (bio)monitoring programs may be a crucial tool to identify firefighters at high risk for developing adverse health outcomes.

2023-04Conference object

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Characterization of the Human Papillomavirus 16 Oncogenes in K14HPV16 Mice: Sublineage A1 Drives Multi-Organ Carcinogenesis

Publication . Cochicho, Daniela; Nunes, Alexandra; Gomes, João Paulo; Martins, Luís; Cunha, Mário; Medeiros-Fonseca, Beatriz; Oliveira, Paula; Bastos, Margarida M.S.M.; Medeiros, Rui; Mendonça, Joana; Vieira, Luis; Gil da Costa, Rui M.; Felix, Ana

The study of ()-induced carcinogenesis uses multiple in vivo mouse models, one of which relies on the cytokeratin 14 gene promoter to drive the expression of all HPV early oncogenes. This study aimed to determine the HPV16 variant and sublineage present in the K14HPV16 mouse model. This information can be considered of great importance to further enhance this K14HPV16 model as an essential research tool and optimize its use for basic and translational studies. Our study evaluated HPV DNA from 17 samples isolated from 4 animals, both wild-type (n = 2) and HPV16-transgenic mice (n = 2). Total DNA was extracted from tissues and the detection of HPV16 was performed using a qPCR multiplex. HPV16-positive samples were subsequently whole-genome sequenced by next-generation sequencing techniques. The phylogenetic positioning clearly shows K14HPV16 samples clustering together in the sub-lineage A1 (NC001526.4). A comparative genome analysis of K14HPV16 samples revealed three mutations to the human papillomaviruses type 16 sublineage A1 representative strain. Knowledge of the HPV 16 variant is fundamental, and these findings will allow the rational use of this animal model to explore the role of the A1 sublineage in HPV-driven cancer.

2022-10-15Journal article

Open access

Portuguese wildland firefighters: assessing exposure and cytogenetic effects in non-fire settings during the pre-fire season

Publication . Pires, Joana; Esteves, Filipa; Slezakova, Klara; Madureira, Joana; Vaz, Josiana; Fernandes, Adília; Pereira, Maria do Carmo; Morais, Simone; Teixeira, João Paulo; Costa, Solange

IARC has recently listed occupational exposure as a firefighter carcinogenic to humans. However, firefighter is among the least studied occupations. The goal of the present study was to assess the level of cytogenetic damage in buccal mucosa (first-contact exposure tissue) of Portuguese firefighters during the pre- wildland fire season, as well as, to evaluate firefighters ́ exposure to particulate matter (PM) on fire stations as a baseline for occupational exposure characterization. A group of 172 Portuguese wildland firefighters were enrolled in the study, relevant information was obtained through a questionnaire. The frequency of micronucleus and other endpoints were evaluated via buccal micronucleus cytome assay (BMCyt) along with the influence of potential risk factors (e.g., smoking habits). Estimated inhalation doses of PM10 and PM2.5 (indoor/outdoor) were estimated for a group of 80 firefighters. No significant associations were found between estimated inhaled doses and BMCyt endpoints. Some lifestyle/behavioural variables were shown to significantly decrease the levels of cytogenetic endpoints, whereas others were found to increase them. Firefighters of the Permanent Intervention Team also shown to have significantly higher levels of BMCyt endpoints compared to other firefighters. Fire seasons are getting longer and dangerous, and this trend will continue in a progressive warming world. Implementing effective mitigation strategies for firefighters’ occupational exposure is an urgent public health issue.

2023-06-21Conference object

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