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- Baseline data and associations between urinary biomarkers of polycyclic aromatic hydrocarbons, blood pressure, hemogram, and lifestyle among wildland firefightersPublication . Barros, Bela; Paiva, Ana Margarida; Oliveira, Marta; Alves, Sara; Esteves, Filipa; Fernandes, Adília; Vaz, Josiana; Slezakova, Klara; Costa, Solange; Teixeira, João Paulo; Morais, SimoneIntroduction: Available literature has found an association between firefighting and pathologic pathways leading to cardiorespiratory diseases, which have been linked with exposure to polycyclic aromatic hydrocarbons (PAHs). PAHs are highlighted as priority pollutants by the European Human Biomonitoring Initiative in occupational and non-occupational contexts. Methods: This cross-sectional study is the first to simultaneously characterize six creatinine-adjusted PAHs metabolites (OHPAHs) in urine, blood pressure, cardiac frequency, and hemogram parameters among wildland firefighters without occupational exposure to fire emissions (> 7 days), while exploring several variables retrieved via questionnaires. Results: Overall, baseline levels for total OHPAHs levels were 2 to 23-times superior to the general population, whereas individual metabolites remained below the general population median range (except for 1-hydroxynaphthalene+1-hydroxyacenaphtene). Exposure to gaseous pollutants and/or particulate matter during work-shift was associated with a 3.5-fold increase in total OHPAHs levels. Firefighters who smoke presented 3-times higher total concentration of OHPAHs than non-smokers (p < 0.001); non-smoker females presented 2-fold lower total OHPAHs (p = 0.049) than males. 1-hydroxypyrene was below the recommended occupational biological exposure value (2.5 μg/L), and the metabolite of carcinogenic PAH (benzo(a)pyrene) was not detected. Blood pressure was above 120/80 mmHg in 71% of subjects. Firefighters from the permanent intervention team presented significantly increased systolic pressure than those who performed other functions (p = 0.034). Tobacco consumption was significantly associated with higher basophils (p = 0.01–0.02) and hematocrit (p = 0.03). No association between OHPAHs and blood pressure was found. OHPAHs concentrations were positively correlated with monocyte, basophils, large immune cells, atypical lymphocytes, and mean corpuscular volume, which were stronger among smokers. Nevertheless, inverse associations were observed between fluorene and pyrene metabolites with neutrophils and eosinophils, respectively, in non-smokers. Hemogram was negatively affected by overworking and lower physical activity. Conclusion: This study suggests possible associations between urinary PAHs metabolites and health parameters in firefighters, that should be further assessed in larger groups.
- Biomonitoring of firefighters’ exposure to priority pollutant metal(loid)s during wildland fire combat missions: Impact on urinary levels and health risksPublication . Paiva, Ana Margarida; Barros, Bela; Azevedo, Rui; Oliveira, Marta; Alves, Sara; Esteves, Filipa; Fernandes, Adília; Vaz, Josiana; Alves, Maria José; Slezakova, Klara; Pereira, Maria do Carmo; Teixeira, João Paulo; Costa, Solange; Almeida, Agostinho; Morais, SimoneWildland firefighters are exposed to metal(loid)s released during wildfires through vegetation combustion, which also promotes remobilization of accumulated anthropogenic metal(loid)s. Studies biomonitoring metal(loid)s exposure promoted exclusively by wildfire suppression activities are lacking. This work aimed to characterize, for the first time, the impact of real-life wildland firefighting operations on urinary levels of priority pollutant metal(loid)s [14 included in ATSDR, 11 in USEPA, and 4 in Human Biomonitoring for Europe Initiative priority lists] in firefighters. Spot urines were sampled pre-exposure (105 non-smokers, 76 smokers) and post-exposure to firefighting activities (20 non-smokers, 25 smokers); among those, paired samples were collected from 14 non-smoking and 24 smoking firefighters. Smokers displayed significantly higher baseline levels of zinc (28 %), lithium (29 %), cadmium (55 %), rubidium (13 %), and copper (20 %) than non-smokers. Following wildfire suppression, the concentration of the WHO potentially toxic metal(loid)s rose from 2 % to 3 % in smokers and 2 % to 5 % in non-smokers (up to 4 % for all firefighters and up to 5 % in paired samples). Levels of nickel (33-53 %), antimony (45-56 %), and cesium (40-47 %) increased significantly post-exposure in non-smokers (in all firefighters and in paired samples), whose urinary concentrations were generally more impacted by wildfire emissions than those of smokers. Arsenic (80 %) displayed the only significant increase post-exposure in smokers, being the best discriminant of exposure to wildfire emissions in these subjects. Significant positive correlations were found for age and/or career length with cadmium, lead, barium, strontium, and mercury, and for body mass index with arsenic. The reference/guidance values were exceeded for arsenic, zinc, cesium, nickel, antimony, cadmium, lead, thallium, mercury, copper, and cobalt in 1-90 % of firefighters suggesting augmented health risks due to wildfire combating and emphasizing the need of mitigation strategies. This study also provides biomonitoring data to help setting reference values for the occupationally exposed part of population.
- Biomonitoring of polycyclic aromatic hydrocarbons exposure and short-time health effects in wildland firefighters during real-life fire eventsPublication . Paiva, Ana Margarida; Barros, Bela; Oliveira, Marta; Alves, Sara; Esteves, Filipa; Fernandes, Adília; Vaz, Josiana; Slezáková; Teixeira, João Paulo; Costa, Solange; Morais, SimoneHuman biomonitoring data retrieved from real-life wildland firefighting in Europe and, also, worldwide are scarce. Thus, in this study, 176 Portuguese firefighters were biomonitored pre- and post- unsimulated wildfire combating (average:12-13 h; maximum: 55 h) to evaluate the impact on the levels of urinary polycyclic aromatic hydrocarbons hydroxylated metabolites (OHPAH; quantified by high-performance liquid chromatography with fluorescence detection) and the associated short-term health effects (symptoms, and total and differentiated white blood cells). Correlations between these variables and data retrieved from the self-reported questionnaires were also investigated. Firefighters were organized into four groups according to their exposure to wildfire emissions and their smoking habits: non-smoking non-exposed (NSNExp), non-smoking exposed (NSExp), smoking non-exposed (SNExp), and smoking and exposed (SExp). The most abundant metabolites were 1-hydroxynaphthalene and 1-hydroxyacenaphthene (1OHNaph + 1OHAce) (98-99 %), followed by 2-hydroxyfluorene (2OHFlu) (0.2-1.1 %), 1-hydroxyphenanthrene (1OHPhen) (0.2-0.4 %), and 1-hydroxypyrene (1OHPy) (0.1-0.2 %); urinary 3-hydroxybenzo(a)pyrene was not detected. The exposure to wildfire emissions significantly elevated the median concentrations of each individual and total OHPAH compounds in all groups, but this effect was more pronounced in non-smoking (1.7-4.2 times; p ≤ 0.006) than in smoking firefighters (1.3-1.6 times; p ≤ 0.03). The greatest discriminant of exposure to wildfire emissions was 1OHNaph + 1OHAce (increase of 4.2 times), while for tobacco smoke it was 2OHFlu (increase of 10 times). Post-exposure, white blood cells count significantly increased ranging from 1.4 (smokers, p = 0.025) to 3.7-fold (non-smokers, p < 0.001), which was accompanied by stronger significant correlations (0.480 < r < 0.882; p < 0.04) between individual and total OHPAH and total white blood cells (and lymphocytes > monocytes > neutrophils in non-smokers), evidencing the impact of PAH released from wildfire on immune cells. This study identifies Portuguese firefighters with high levels of biomarkers of exposure to PAH and points out the importance of adopting biomonitoring schemes, that include multiple biomarkers of exposure and biomarkers of effect, and implementing mitigations strategies.
- Cumulus cell DNA damage linked to fertilization success in females with an ovulatory dysfunction phenotypePublication . Rodrigues, Bárbara; Sousa, Vanessa; Esteves, Filipa; Vale-Fernandes, Emídio; Costa, Solange; Sousa, Daniela; Brandão, Raquel; Leal, Carla; Pires, Joana; Gaivão, Isabel; Teixeira, João Paulo; Nogueira, António J.A.; Jorge, PaulaIntracytoplasmic sperm injection (ICSI) is a widely used technique in fertility centers. ICSI success depends on both nuclear and cytoplasmic oocyte maturation. Cumulus cells, which surround the oocytes, play a pivotal role in oocyte competence. However, the significance of DNA damage in cumulus cells as a marker of fertilization success remains largely unexplored. This study aims to investigate the relationship between DNA damage in cumulus cells of females undergoing ICSI, and oocyte competence, with a focus on in vitro fertilization (IVF) outcomes. We employed the alkaline comet assay to assess DNA damage levels (%TDNA) in cumulus cells and whole blood from 22 potentially fertile females and 35 infertile females, including 20 with an ovulatory disfunction phenotype. Our results revealed significant differences between the levels of %TDNA in cumulus cells and blood. Females with an ovulatory dysfunction phenotype exhibited higher levels of %TDNA in cumulus cells compared to potentially fertile females. Additionally, within the group of females with ovulatory dysfunction, a significant correlation was observed between %TDNA levels and the number of oocytes with two pronuclei. Our findings suggest that blood does not accurately reflect DNA damage in cumulus cells, which was correlated with the fertilization success in females with ovulatory dysfunction. High levels of %TDNA in cumulus cells were associated with a higher likelihood of successful fertilization. Moreover, our results imply that low levels of %TDNA may be linked to oocytes that fail to complete maturation and, consequently, do not fertilize (oocytes with zero pronuclei). Further research with larger cohorts is necessary to validate these findings and to explore potential applications in female fertility. However, our study provides evidence that DNA damage in cumulus cells may serve as a valuable biomarker for predicting fertilization success and oocyte competence.
- FAIREHR: A Novel Online Research Registry PlatformPublication . Galea, Karen S.The FAIREHR (Findable, Accessible, Interoperable, Reusable Environmental and Health Registry) is a state-of-the-art online registry platform designed to enhance the transparency, reproducibility, and comparability of environmental health research, focusing on human biomonitoring (HBM) studies as a starting point. This platform is developed in response to the Europe Regional Chapter of the International Society of Exposure Science (ISES Europe) HBM working group strategic objectives to generate high quality HBM by harmonising the data life cycle and implementation of FAIR (findable, accessible, interoperable and reusable) guiding principles. The registry enables preregistration of studies, capturing key metadata on study design, data management, and planned methods before recruitment od the participants. FAIREHR is the first registry tailored for HBM studies and is also first registry promoting FAIR by design studies. Benefits of FAIREHR include increased research visibility, improved data comparability, enhanced collaboration, and better-informed decision-making. We will discuss the unique propositions of FAIREHR, emphasizing its role in enhancing the exchange of information, with its implementation expected to yield significant benefits for researchers, policymakers, and public health through effective utilization of HBM data.
- Firefighters’ Occupational Exposure in Preparation for Wildfire Season: Addressing Biological ImpactPublication . Esteves, Filipa; Slezakova, Klara; Madureira, Joana; Vaz, Josiana; Fernandes, Adília; Morais, Simone; Pereira, Maria do Carmo; Teixeira, João Paulo; Costa, SolangeThe characterization of wildland firefighters’ occupational exposure must consider different exposures, including those at the fire station. The present study aimed to characterize the occupational exposure of 172 Northern Portuguese wildland firefighters in fire stations during the pre-wildfire season of 2021. The biological impact of estimated inhaled doses of PM10 and PM2.5 (indoor/outdoor) was accessed through a buccal micronucleus cytome (BMCyt) assay in exfoliated buccal cells of a subgroup of 80 firefighters. No significant association was found between estimated inhaled doses of PM10 and PM2.5 (mean 1.73 ± 0.43 µg kg−1 and 0.53 ± 0.21 µg kg−1, respectively) and biological endpoints. However, increased frequencies of cell death parameters were found among subjects of the Permanent Intervention Teams (full-time firefighters). The intake of nutritional supplements was associated with a significant decrease in micronucleus frequencies (i.e., DNA damage or chromosome breakage). In addition, our findings showed a significantly increased frequency of cell death endpoints (i.e., nuclear fragmentation) with coffee consumption, while daily consumption of vegetables significantly decreased it (i.e., nuclear shrinkage). Our results provide data on the occupational exposure of wildland firefighters while working in fire stations during the pre-wildfire season, providing the essential baseline for further studies throughout the wildfire season.
- Guiding principles for mixture threshold derivation from effect biomarkersPublication . OECD Environment Directorate Chemicals and Biotechnology Committee; OECD Environment Health and Safety Division__APAGARForeword: Currently available assessment approaches for cumulative risk of chemical mixtures can only be applicable to a small number of substances present at workplaces and in the environment. We cannot anticipate a significant change of this situation in the near future due to extensive data need for cumulative risk assessments. Presently, effect biomarkers are the most direct option to address the risk of known and unknown mixtures in an integrative way. Traditional occupational health risk assessments often rely on external exposure measurements, such as air monitoring, which may not fully capture the complexities of workplace exposures. Human biomonitoring is used to measure internal exposures or effects in exposed individuals or groups from all potential routes of exposure (i.e., inhalation, oral, and dermal). Exposure to mixtures in the workplace and environment is the most common chemical exposure scenario in our daily lives. However, methods for assessing the risks and for setting mixture threshold limits to avoid adverse effects lack global harmonization. Monitoring of effect biomarkers can support regulatory risk assessment in multiple ways. An effect biomarker indicates a stressor-induced biological effect which can be associated with a disease and can be interpreted as a potential predictor of a downstream effect i.e. measuring a key event in a Mode of Action (MoA) or Adverse Outcome Pathway (AOP). Thus, biomarkers can provide an integrated measure of the response to relevant stressors by all routes of known and unknown exposures. However, effect biomarker responses are usually not straightforward to interpret regarding their predictive value to indicate adverse effects. A systematic understanding of the relevance of effect biomarker data will enhance the protection of workers and/or ecosystems, if used under appropriate ethical and regulatory frameworks. Therefore, harmonized guidance for assessing effect biomarkers and their application to risk assessments are needed. The guiding principles proposed in this document describe the key concepts for the derivation and interpretation of mixture thresholds* for selected effect biomarkers for use in occupational or ecological risk assessments. The aim of these guiding principles is to present a harmonized assessment approach which will save resources and promote consistency across regulatory agencies at national and international levels. The development of this document was a joint activity of the Organisation for Economic Co-operation and Development (OECD) Working Party on Exposure Assessment & Working Party on Hazard Assessment (WPEA & WPHA) in collaboration with more than 90 experts from 25 countries and other stakeholders (see chapter 8 project participation). The activity was started in October 2022 and the development of this guiding principles document was co-led by Robert Pasanen-Kase (SECO*, CH) as coordinator, Maryam Zare-Jeddi (BIAC*), Nancy B. Hopf (Unisanté, CH), Susana Viegas (ENSP*/UNL, PT), Dan Villeneuve (US-EPA*, US), Martin Wilks and Rex FitzGerald (University of Basel, CH), Radu Corneliu Duca (LNS*, LU) and the OECD Secretariat. The document was drafted in close collaboration with experts providing input on different aspects of human and environmental effect-biomonitoring including Bernice Scholten, (TNO* , NL), Eszter Simon (FOEN* , CH), Devika Poddalgoda (Health Canada, CAN), Anna Bal Price (JRC*, EU); Vicente Mustieles, Antonio Hernandez-Jerez (University of Granada, ES), Christoph van Thriel (IFADO* , DE), Stefano Bonassi (IRCCS*, San Raffaele Roma, IT), Michael Fenech (University of South Australia, AUS), Sophie Ndaw (INRS*, FR), Christina Pieper (German Federal Institute for Risk Assessment, DE), Lucian Farcal, Alicia Paini (EFSA*, EU). The initial draft guidance document was reviewed in 2025 by expert group (see chapter 8) and WPEA & WPHA members and was commented by eleven experts from six different organisations / institutes / companies and was finalized. This adopted biomonitoring guiding principles document is published under the responsibility of the Chemical and Biotechnology Committee of the OECD.
- The Impact of Sleep on Haematological Parameters in FirefightersPublication . Alves, Sara; Silva, Francisca; Esteves, Filipa; Costa, Solange; Slezakova, Klara; Alves, Maria; Pereira, Maria; Teixeira, João Paulo; Morais, Simone; Fernandes, Adília; Queiroga, Felisbina; Vaz, JosianaSleep is a vital process that impacts biological functions such as cell renewal, bone regeneration, and immune system support. Disrupted sleep can interrupt erythropoiesis, leading to fewer red blood cells, reduced haemoglobin concentration, and decreased haematocrit levels, potentially contributing to haematological disorders. This is particularly concerning for shift workers for example firefighters. While previous studies have explored sleep’s adverse effects on various professions, research specific to firefighters is limited. This study investigates the relationship between sleep quality and haematological parameters among firefighters in Northeast Portugal. From a sample of 201 firefighters, variations in red blood cells, haemoglobin, and haematocrit values were linked to sleep quality. The study utilised non-parametric tests (Wilcoxon-Mann-Whitney, Spearman’s correlation) to explore the connection between sleep quality and haematological profile. The impact of covariates on haematological parameters was assessed using non-parametric ANCOVA (Quade’s). A multiple regression analysis was employed to further understand how sleep quality and various confounding variables impact haematological levels. Findings suggest a negative link between sleep quality and haematological levels, meaning that as sleep quality deteriorates, there is a tendency for haematological levels to decrease, as indicated by Spearman’s correlation (rRBC = −0.157, pRBC = 0.026; rHb = −0.158, pHb = 0.025; rHCT = −0.175, pHCT = 0.013). As observed in scientific literature, the correlation found suggests a possible inhibition of erythropoiesis, the process responsible for red blood cell production. Despite firefighters presenting a haematological profile within the reference range (RBC: 5.1 × 106/mm3 (SD ± 0.4), Hb: 15.6 g/dL (SD ± 1.3), 47% (SD ± 1.0), there is already an observable trend towards lower levels. The analysis of co-variables did not reveal a significant impact of sleep quality on haematological levels. In conclusion, this study underscores the importance of sleep quality in determining haematological parameters among firefighters. Future research should investigate the underlying mechanisms and long-term implications of poor sleep quality on firefighter health. Exploring interventions to enhance sleep quality is vital for evidence-based strategies promoting firefighter well-being.
- MIR-HBM: The Minimum Information Requirements Guidance for Human Biomonitoring StudiesPublication . Hopf, Nancy B.Background To fully understand the context, methods, data, and interpretations of a human biomonitoring (HBM) study, access to comprehensive background information is essential. However, the diversity in HBM study designs, coupled with varying levels of detail in the data collected, often makes meaningful comparisons, data reuse, and interpretation across different studies challanging. Materials and methods To address this need, the Minimum Information Requirements for Human Biomonitoring (MIR-HBM) was developed by the European Chapter of the International Society of Exposure Science (ISES Europe) HBM working group and the HBM Global Network. MIR-HBM describes the minimal set of information that must be provided to enable effective communication of the setup of an HBM study to others. It enables the generation of the metadata ('data about the data') that will cover all components of the study, including rationale and objectives, study population characteristics, biological specimen collection, laboratory analysis and method parameters, data analysis plan, data interpretation, and communication and reporting. Results Adherence to these reporting guidelines will result in publications of increased clarity,quality, comparability, and usefulness to the scientific community and other stakeholders. Integrating MIR-HBM as a FAIR metadata schema into the FAIREHR (Findable, Accessible, Interoperable, and Reusable Environmental and Health Registry) platform is the next key step planned to enable its implementation and adoption. Conclusions Overall, the MIR-HBM on HBM study metadata promotes transparency and completeness in reporting and enhances rapid capturing of the contents of the HBM study, thereby stimulating findability and accessibility to HBM data and supporting effective quality assessment.
- Occupational exposure to wildland firefighting and its effects on systemic DNA damagePublication . Esteves, Filipa; Madureira, Joana; Costa, Carla; Pires, Joana; Barros, Bela; Alves, Sara; Vaz, Josiana; Oliveira, Marta; Slezakova, Klara; Fernandes, Adília; Pereira, Maria do Carmo; Morais, Simone; Valdiglesias, Vanessa; Bonassi, Stefano; Teixeira, João Paulo; Costa, SolangeBackground: Portugal is among the European Union countries more devastated by forest fires. Wildland firefighters are at the forefront of this battle, facing exposure to a wide range of harmful pollutants. Epidemiological studies have highlighted a potential link between occupational firefighting exposure and several diseases, including cancer. To date, very few studies have explored the biological mechanisms associated with such exposure. The present longitudinal study aims to assess changes in early effect biomarkers following wildland firefighters’ occupational exposure to a real wildfire event. Methods: Paired blood samples from 59 healthy Portuguese wildland firefighters were collected at two different time points: before wildfire season and after a fire event during wildfire season. Sociodemographic variables (e.g., age, sex) and work-related factors (e.g., years of service) were assessed via a self-reported questionnaire. Levels of early effect biomarkers, such as primary DNA damage and oxidative DNA damage (oxidised purines) were assessed via comet assay. DNA double-strand breaks (DSBs) were evaluated by phosphorylated H2AX (γH2AX). Moreover, hydroxylated polycyclic aromatic hydrocarbon metabolites (OHPAHs) and metal(loid)s were quantified in urine samples. The influence of urinary OHPAHs, urinary metal(loid)s, and other exposure-related factors (e.g., firefighting duration) on changes (Δ) in early effect biomarkers (post-vs. baseline levels) was investigated. Results: Firefighting activities led to a significant increase in both primary DNA damage and oxidative DNA damage by 22 % (95 % CI: 1.11–1.35; p < 0.05) and 23 % (95 % CI: 1.04–1.45; p < 0.05), respectively. Results from linear regression revealed that per each unit increase of urinary 2-hydroxyfluorene (2-OHFlu) (μmol/mol creatinine), the risk of ⧍ oxidative DNA damage increased by 20 % [FR: 1.20 (1.09–1.32); p < 0.01]. Additionally, each unit increase in urinary cesium (Cs) (μg/L) resulted in a significant 4 % increase in Δ primary DNA damage [FR: 1.04 (1.01–1.06); p < 0.05] and a 3 % increase in Δ oxidative DNA damage [FR: 1.03 (1.01–1.05); p < 0.05]. Post-exposure levels of γH2AX were significantly correlated with urinary 2-OHFlu levels assessed after firefighting (r = 0.30; p < 0.05). Furthermore, exposure duration and reported breathing difficulties during firefighting were significantly associated with increased levels of primary DNA damage. Conclusion: Results obtained provide insights into the potential human health effects of wildland firefighting occupational exposure at the genetic and molecular levels, offering new and important mechanistic data. These findings are crucial for implementing health and safety measures, recommendations, and best practices to mitigate occupational risks and protect the health of wildland firefighters.