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- Complementary field and laboratory batch studies to quantify generation rates of perfluoroalkyl acids in a contaminated agricultural topsoil with unknown precursorsPublication . Haluska, Alexander Arthur; Röhler, Klaus; Fabregat-Palau, Joel; Alexandrino, Diogo A. M.; Abramov, Sergey; Thompson, Katharine J.; Straub, Daniel; Kleindienst, Sara; Bugsel, Boris; Zweigle, Jonathan; Zwiener, Christian; Grathwohl, Peter; Alexandrino, DiogoSoil microbiome changes and generation rates of per- and polyfluoroalkyl substances (PFAS) precursors were studied in a contaminated agricultural field using a combination of field and laboratory batch microcosm studies. 16S rRNA gene amplicon sequencing was used to track how microbial community composition changed over time, while perfluoroalkyl acids (PFAA) generation rates were quantified using a combination of field and batch incubations combined with the direct total oxidizable precursor (dTOP) assay. The study site in Brilon-Scharfenberg, North Rhine-Westphalia, Germany, has PFAS contamination in the topsoil (0 to 30cm) originating from compost. Generation rate constants of these short-chain PFAA estimated from batch incubations (0.12 to 0.751/year) were higher but similar to rate constants from the fields (0.05 to 0.221/year). Long-term field mass discharge data (2009 to 2023) suggest that at least 60years are needed to remove 99.99% of short-chain PFAA and their precursors. 16S rRNA gene amplicon sequencing data revealed a major impact of PFAA on the biodiversity of soil microorganisms, with batch-incubated contaminated soils showing higher richness and diversity indexes than field control soils. However, most of these impacts occurred at lower taxonomical ranks and did not seem to have a prominent impact on the overall structure of the autochtonous microbial communities of the soils where PFAA were produced and accumulated. Overall, our findings demonstrate that well-controlled aerobic batch test combined with the results of dTOP assay are a suitable approach for estimating short-chain PFAA generation rates. Additionally, our research suggests that the complete removal of PFAA precursors from topsoil will take decades.
- Microcystin-degrading bacteria reduce bioaccumulation in Fragaria vulgaris and enhance fruit yield and qualityPublication . Haida, Mohammed; Khalloufi, Fatima El; Essadki, Yasser; Alexandrino, Diogo A. M.; Mugani, Richard; Hejjaj, Abdessamad; Campos, Alexandre; Vasconcelos, Vitor; Carvalho, Maria F.; Díez‑Quijada, Leticia; Cameán, Ana M.; Oudra, Brahim; Alexandrino, DiogoIn Morocco, red fruit production has thrived, primarily utilizing hydroponic methods to control crops, increase fruit yield and quality, and avoid soil-related problems. However, the irrigation of these expansive hydroponic farms relies heavily on water sourced from dams, many of which are contaminated with Microcystins (MCs). To address this contamination issue, ongoing research is focused on discovering effective and cost-efficient biological solutions for eliminating MCs. In this study, we isolate and identify bacterial strains capable of degrading MCs, evaluate the rate of degradation, and investigate how soil inoculated with these bacteria affects the accumulation of MCs in plant tissue. The partial 16S rRNA analyses of three bacterial sequences were conducted, identifying them through NCBI as follows: Ensifer sp. (B1) isolated from soil, Shinella sp. (B2) from a cyanobacterial bloom, and Stutzerimonas sp. (B3) from water. These bacteria exhibited the ability to degrade MCs, with approximately 34.75%, 73.75%, and 30.1% of the initial concentration (20 µg/L) being removed after a 6-day period for B1, B2, and B3, respectively. Moreover, strawberry plants were cultivated hydroponically in a greenhouse for a duration of 90 days. These plants were subjected to extracts of cyanobacteria containing 10 and 20 µg/L of Microcystins (MC), as well as water from an artificial lake contaminated with MC, both with and without the presence of isolated bacterial strains. Among these strains, Shinella sp. exhibited the highest efficacy in mitigating MC accumulation. Specifically, it resulted in a reduction of approximately 1.159 µg of MC per kilogram of root dry weight, leading to complete elimination in the leaves and fruits. The findings also indicated that the inoculation of perlite with the three MC-degrading bacterial strains significantly enhanced growth, photosynthetic pigments, yield, biochemical constituents, and quality attributes of strawberries (p≤0.05). These promising outcomes suggest the potential of this approach for addressing the adverse impacts of crops irrigated with MC-contaminated water in future agricultural practices.
- Unveiling the culturable and non‐culturable actinobacterial diversity in two macroalgae species from the northern Portuguese coastPublication . Girão, Mariana; Alexandrino, Diogo A. M.; Cao, Weiwei; Costa, Isabel; Jia, Zhongjun; Carvalho, Maria F.; Alexandrino, DiogoActinomycetota, associated with macroalgae, remains one of the least explored marine niches. The secondary metabolism of Actinomycetota, the primary microbial source of compounds relevant to biotechnology, continues to drive research into the distribution, dynamics, and metabolome of these microorganisms. In this study, we employed a combination of traditional cultivation and metagenomic analysis to investigate the diversity of Actinomycetota in two native macroalgae species from the Portuguese coast. We obtained and taxonomically identified a collection of 380 strains, which were distributed across 12 orders, 15 families, and 25 genera affiliated with the Actinomycetia class, with Streptomyces making up approximately 60% of the composition. Metagenomic results revealed the presence of Actinomycetota in both Chondrus crispus and Codium tomentosum datasets, with relative abundances of 11% and 2%, respectively. This approach identified 12 orders, 16 families, and 17 genera affiliated with Actinomycetota, with minimal overlap with the cultivation results. Acidimicrobiales emerged as the dominant actinobacterial order in both macroalgae, although no strain affiliated with this taxonomic group was successfully isolated. Our findings suggest that macroalgae represent a hotspot for Actinomycetota. The synergistic use of both culture-dependent and independent approaches proved beneficial, enabling the identification and recovery of not only abundant but also rare taxonomic members.
- Impact of per- and polyfluorinated alkyl substances (PFAS) on the marine environment: Raising awareness, challenges, legislation, and mitigation approaches under the One Health conceptPublication . Bilela, Lada Lukić; Matijošytė, Inga; Krutkevičius, Jokūbas; Alexandrino, Diogo A.M.; Safarik, Ivo; Burlakovs, Juris; Gaudêncio, Susana P.; Carvalho, Maria F.; Alexandrino, DiogoPer- and polyfluorinated alkyl substances (PFAS) have long been known for their detrimental effects on the ecosystems and living organisms; however the long-term impact on the marine environment is still insufficiently recognized. Based on PFAS persistence and bioaccumulation in the complex marine food network, adverse effects will be exacerbated by global processes such as climate change and synergies with other pollutants, like microplastics. The range of fluorochemicals currently included in the PFAS umbrella has significantly expanded due to the updated OECD definition, raising new concerns about their poorly understood dynamics and negative effects on the ocean wildlife and human health. Mitigation challenges and approaches, including biodegradation and currently studied materials for PFAS environmental removal are proposed here, highlighting the importance of ongoing monitoring and bridging research gaps. The PFAS EU regulations, good practices and legal frameworks are discussed, with emphasis on recommendations for improving marine ecosystem management.
- Nano versus bulk: Evaluating the toxicity of lanthanum, yttrium, and cerium oxides on Enchytraeus crypticusPublication . Santos, Joana; Neca, Inês; Capitão, Ana; Nogueira, João; Santos, José A. S.; Pinto, Edgar; Barreto, Angela; Silva, Ana L. Daniel da; Maria, Vera L.; Pinto, EdgarConsidering the increase in demand for rare earth elements (REEs) and their accumulation in soil ecosystems, it is crucial to understand their toxicity. However, the impact of lanthanum, yttrium and cerium oxides (La2O3, Y2O3 and CeO2, respectively) on soil organisms remains insufficiently studied. This study aims to unravel the effects of La2O3, Y2O3 and CeO2 nanoparticles (NPs) and their corresponding bulk forms (0, 156, 313, 625, 1250 and 2500 mg/kg) on the terrestrial species Enchytraeus crypticus. The effects on survival, reproduction (21 days (d)), avoidance behavior (2 d) and DNA integrity (2 and 7 d) of E. crypticus were evaluated. No significant effects on survival were observed. For La2O3, the bulk form affected more endpoints than the NPs, inducing avoidance behavior (1250 mg/kg) and DNA damage (1250 mg/kg - 2 d; 2500 mg/kg - 7 d). The Y2O3 NPs demonstrated higher toxicity than the bulk form: decreased reproduction (≥ 1250 mg/kg); induced avoidance behavior (≥ 625 mg/kg) and DNA damage (≥ 156 mg/kg - 2 d; 2500 mg/kg - 7 d). For CeO2, both forms exhibited similar toxicity, decreasing reproduction (625 mg/kg for bulk and 2500 mg/kg for NPs) and inducing DNA damage at all tested concentrations for both forms. REEs oxides toxicity was influenced by the REEs type and concentration, exposure time and material form, suggesting different modes of action. This study highlights the distinct responses of E. crypticus after exposure to REEs oxides and shows that REEs exposure may differently affect soil organisms, emphasizing the importance of risk assessment.
- Evaluation of the potential of annatto seed powder to reduce the formation of heterocyclic amines in charcoal-grilled and pan-fried beef pattiesPublication . Gonçalves, Thaís de Moura Neves; Braga, Anna Rafaela Cavalcante; Pinto, Edgar; Viegas, Olga; Mesquita, Leonardo M. de Souza; Ferreira, Isabel Maria Pinto Leite Viegas Oliveira; García-Jares, Carmen; De Rosso, Veridiana Vera; Domene, Semíramis Martins Álvares; Pinto, EdgarVarious strategies are being explored to reduce the formation of undesirable compounds during the thermal processing of foods. This study investigates the impact of incorporating annatto seed powder (Bixa orellana L.) into beef patties to reduce the formation of heterocyclic amines (HAs) during charcoal-grilling and pan-frying. A three-level full factorial design was used to assess the effect of both annatto seed powder concentration and cooking times on HAs formation. The results showed that HA formation increased with longer cooking times and decreased with higher concentrations of annatto seed powder. A significant reduction in HA content was observed in both charcoal-grilled and pan-fried beef patties when annatto seed powder was added, with a particularly notable 91 % reduction at the 1 % addition level. These findings demonstrate that the addition of annatto seed powder is a highly effective strategy for reducing HA formation in beef patties. Chemical compounds studied in this article: 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) (PubChem CID: 62275); 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline (4,8-DiMeIQx) (PubChem CID: 104739); 2amino-3,7,8-trimethylimidazo[4,5-f]quinoxaline (7,8-DiMeIQx) (PubChem CID: 104855); 2-amino-1-methyl-6phenylimidazo[4,5-b]pyridine (PhIP) (PubChem CID: 1530); 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (TrpP-1) (PubChem CID: 5284474); 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2) (PubChem CID: 5284476); 2-amino-9H-pyrido[2,3-b]indole (A α (MeA α C) (PubChem CID: 62805); 2-amino-3-methyl-9H-pyrido[2,3-b]indole C) (PubChem CID: 62244); Bixin (PubChem CID: 5281226).
- Stock structure of the gulf hake urophycis cirrata (Teleostei: Phycidae) in south-western atlantic using Otolith shape and elemental analysesPublication . Santificetur, César; Rossi-Wongtschowski, Carmen Lúcia Del Bianco; Ruperti, André; Almeida, Agostinho; Pinto, Edgar; Correia, Alberto Teodorico; Pinto, EdgarUrophycis cirrata is an important demersal fish species targeted by Brazilian industrial fisheries. With high exploitation rates, its stock(s) is(are) currently deemed fully exploited or overexploited. While basic ecological information, such as length at first maturity, exists, knowledge of its population structure is limited. A sub-sample of 90 sagittal otoliths of U. cirrata juveniles (300–411 mm total length) collected during the Program for Assessment of the Sustainable Potential of Living Resources in the Exclusive Economic Zone (REVIZEE) in 2001/2002 was analyzed. Samples came from the outer continental shelf and upper slope of the southeast-south Brazilian coast, divided into three regions: northern (Cabo São Tomé to São Sebastião), central (São Sebastião to Cabo Santa Marta Grande), and southern (Cabo Santa Marta Grande to Chuí). Otolith shape (elliptic Fourier descriptors) and elemental (element:Ca) signatures were examined using univariate (ANOVA, Tukey) and multivariate (MANOVA, LDFA) statistical methods. An overall reclassification success rate of 86% was achieved using both signatures. However, individuals from the three regions were not fully separable, indicating a single, albeit not homogeneous, population unit for fisheries management. As fish stocks are dynamic, contemporary studies should be conducted to verify whether this population structure persists.
- Assessing work-related musculoskeletal disorders and psychosocial risks in bus drivers: Insights from a municipal company case study in PortugalPublication . Silva, Tânia T.; Mendes, Tatiana R.; Lapa, Inês; Carvalho, Paulo; Rodrigues, Matilde A.; Rodrigues, Matilde; Carvalho, PauloThe public transport sector plays a crucial role in society, oering essential services and providing employment to a significant number of drivers. Despite the importance of this sector, it is essential to recognize that drivers are exposed to various occupational risks inherent to their daily work, which can have serious implications for their health. This study aims to characterize and analyse Work-Related Musculoskeletal Disorders (WMSD) and psychosocial risks in a public transport company. In the initial phase of the study, a questionnaire was administered to assess musculoskeletal symptoms and psychosocial risks. In the second phase, an inertial motion capture system was used to evaluate the risk of developing WMSD. The results revealed a significant and concerning prevalence of burnout, with over 60% of workers reporting high or severe levels across all dimensions (i.e., personal, work-related, and client-related burnout). Depression, anxiety, and stress were within typical ranges, though a relevant percentage of participants exhibited severe and extremely severe levels of depression (7.2%), anxiety (12.2%), and stress (8%). Musculoskeletal discomfort was highly prevalent, particularly in the lower back (68.3%) and neck regions (57.2%), regarding pain over the last 12 months. Additionally, the risk of developing WMSDs was high across the various microtasks, which were analyzed across dierent bus lines and routes, with Rapid Upper Limb Assessment (RULA) scores ranging from 4 (Medium Risk) to 7 (Very High Risk). Based on the results, varying bus types and routes is recommended. Programs should enhance wellbeing, and studies should assess interventions on health, stress, and occupational risks focused on enhancing worker wellbeing should be implemented, and future studies should assess the impact of interventions targeting health, stress, and occupational risks.
- Colouring applications of microalgae and cyanobacteria photosynthetic pigments: Challenges for industrial and market acceptancePublication . Reimão, Mariana; Almeida, Liliana; Ramos, Cristiana; Eusébio, Nádia; Martins, Rosário; Silva, Manuela Vieira da; Vasconcelos, Vítor; Freitas, Marisa; Freitas, Marisa; Vieira da Silva, Manuela; Martins, Rosario; REIMÃO BORGES LOPES DA SILVA, MARIANAMicroalgae and cyanobacterial photosynthetic pigments possess immense potential for sustainable colouring applications in industries such as food, feed, cosmetics and textiles, primarily because of their vibrant colours, safety and eco-friendliness. These photosynthetic pigments align with the growing demand for eco-conscious products. However, substantial challenges persist at the industrial level, particularly in scaling up production and achieving acceptance in niche markets. This review highlights the critical roles of abiotic factors such as light quality, temperature, pH, nutrient availability and salinity in enhancing pigment production using conventional production methods. Additionally, issues related to the cost, purity and stability of the pigments must be addressed to facilitate broader acceptance. Looking forward, the focus must be on developing innovative cultivation techniques that optimise and define the abiotic conditions most favourable for maximum pigment production or even incorporate genetic engineering tools. Additionally, exploring untapped species from inhospitable environmental sources and integrating the residual biomass through biorefinery processes represent effective approaches for enhancing the sustainability of microalgae and cyanobacterial pigment production on an industrial scale. Overcoming these barriers utilising targeted research and industry collaboration as well as fostering market acceptance is essential for fully harnessing the potential of these natural pigments and driving the transition towards more sustainable industrial colouring practices.
- Prevalence of the use of dosimeters for ionizing radiation from fluoroscopy − a systematic literature review and meta-analysisPublication . Videira, Sara; Rodrigues, Matilde; Santos, Joana; Guedes, Joana; Martins, João; Silva, Manuela Vieira da; Vieira da Silva, Manuela; Rodrigues, MatildeThis study aims to assess the prevalence of individual dosimeter use among workers exposed to ionizing radiation during fluoroscopy-guided procedures. Additionally, factors contributing to its use were identified. Studies were identified through searches in five databases on 13 April 2024. Additionally, snowballing techniques were employed. The review followed PRISMA guidelines and the CoCoPop model. A narrative synthesis, bibliometric analysis, and meta-analysis were performed. Study quality was assessed using the Joanna Briggs Institute checklist for prevalence studies. Fifty studies involving 11,067 individuals were included. Orthopedics/traumatology was the most studied specialty (46 %). Median use rates were: 24 %(IQR = 44 %) for eye lens dosimeters, 15 %(IQR = 13 %) for electronic real-time dosimeters, 27 %(IQR = 42 %) for wrist/finger dosimeters, 25 %(IQR = 23 %) for collar/ thyroid dosimeters and 5 % for ankle dosimeter; 15 %(IQR = 78 %) reported no monitoring device. In 15 studies (30 %) with 4,188 individuals (38 %), the overall prevalence of “always” using whole-body dosimeters was 43 % [95 %CI:24–62]. By continent, the highest prevalence was found in Africa (75 %[95 % CI: 46–95]), while the lowest was in the “Americas” (33 %[95 % CI: 16–52]). Significant moderator effects were found: higher prevalence in “Africa” (p = 0.04), “gastroenterology” (p = 0.04), and “involving radiology” (p = 0.01); lower in “orthopedics” (p = 0.01) and “physicians” (p = 0.03). No significant moderator effects were found: “very high Human Development Index” (p = 0.72) and “high Human Development Index” (p = 0.69). Studies showed moderate risk of bias (6/9), with little evidence of publication bias. Exposure doses may be underestimated due to the low prevalence of dosimeter use. Interventions targeting individual and organizational factors are needed to promote consistent use and improve safety.