CIMO - Resumos em Proceedings Não Indexados à WoS/Scopus
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- Adaptive genomic variation in honey bees from arid regionsPublication . Yadró Garcia, Carlos A.; Henriques, Dora; Haddad, N.; Obeidat, W. M.; AlShagour, B.; Muz, Mustafa Necati; Arab, A.; Eissa, A. A.; Hosri, C.; Lamghari, F.; Arruda, J.; Rufino, José; Martín Hernández, R.; Nanetti, A.; Pinto, M. AliceThe Middle East is home to a large number of Apis mellifera subspecies that have evolved under extreme aridity and desertic conditions. Therefore, they represent a unique opportunity to disentangle the genetic basis of adaptation to such challenging environmental conditions. We arranged a collection of 514 drone samples (haploid males) split into two latitudinal transects. Transect 1 comprised 342 samples: 66 A. m. lamarckii (Egypt), 197 A. m. syriaca (Jordan), 30 A. m. syriaca (Lebanon), 21 A. m. meda (southeastern Turkey), and 27 A. m. anatoliaca (Anatolia, Turkey). Transect 2 comprised 173 samples: 14 A. m. jemenitica (Oman), 9 A. m. jemenitica (UAE), 75 A. m. meda (Iran), and 75 A. m. caucasia (northeastern Turkey). For all samples, whole genomes were resequenced, and environmental and bioclimatic variables were retrieved from WorldClim. For each transect, we applied three genotype–environment association methods (Samβada, pRDA, and LFMM) and one outlier-detection approach (PCAdapt) to identify SNPs associated with environmental adaptation. SNPs were annotated, and the intersection of genes between the four methods in each transect was analysed for gene enrichment. In both transects, enriched terms included genes related to biological regulation through endocrine, neuroendocrine, and neurotransmitter pathways. The regulation of gene expression was also enriched, including several transcription factors, especially HOX genes. Transmembrane and ionic transporters also seem to play a central role in environmental adaptation in these subspecies. Finally, neurogenesis, synapse establishment, and neural system development were also enriched. When gene enrichment analysis was set to exclude electronic annotations (automatically assigned, non-curated GO terms), enriched terms included processes such as behaviour, learning or memory, cognition, and associative and olfactory learning and behaviour. Our findings point to a complex suite of regulatory and sensory 62 pathways that collectively shape the environmental adaptation of honey bee subspecies that have evolved in arid environments.
- Application of a lab-made electronic nose as a tool to assess extra virgin olive oil sensory categoryPublication . Ferreiro, Nuno Manuel; Veloso, Ana C.A.; Rodrigues, Nuno; Pereira, José Alberto; Peres, António M.Premium extra virgin olive oils are usually subjected to national and international contests aiming to promote their competitiveness and internationalization capacity. Olive oils were assessed by a sensory expert panel and initially split according to the perceived intensity of the fruitiness sensation into four categories, namely ripe (RF), green delicate (GD), green medium (GM), and green robust (GR) fruitiness.1 However, even for trained panelists, the assessment comprises a subjectivity degree, and the number of oils that can be evaluated per day is limited. In this sense, the use of electronic sensing devices can be helpful as pre-assessment tools, which may allow reducing the initial number of oils to be evaluated. Thus, a lab-made electronic nose (E-nose) comprising nine commercial metal oxide semiconductor (MOS) sensors, previously developed by the research team,2 was applied (Figure 1) aiming to verify its suitability for this purpose. In total, 59 olive oils were included in this study, which were previously classified by an expert sensory panel in a national contest: 20 oils as RF, 15 as GD, 17 as GM and 7 as GR fruitiness. For the E-nose analysis, 0.5 mL of each olive oil was inserted into a 25 mL glass vial and placed in the sampling chamber at 28 ºC (temperature recommended by the International Olive Council for sensory analysis of olive oils) for 13-min, allowing to generate a volatile fraction representative of the sample. After a cleaning step of the sensors’ surfaces using an air flow, the gas headspace from each sample was directed into the detection chamber, where it interacted with the MOS sensors for 2.5 min. The resistance signals of each of the nine MOS sensors were recorded by a data logger at 4 sec intervals being then treated taking into account seven distinct feature extraction methods: the last response point (LP), the integral of the response curve (INT), the maximum response point (MAX), the minimum response point (MIN), the sum of the response curve (SUM), and the mean of the response curve (MEAN).3 The results showed that the E-nose feature extracted data could be used to satisfactorily discriminate, based on a linear discriminant analysis (LDA) coupled with a simulated annealing (SA) algorithm, the olive oils according to sensory category groups with a sensitivity of 100% for training (Figure 2a) and 73% for the leave-one-out cross-validation (LOO-CV) procedure. The lower sensitivity achieved for the internal validation was mainly due to misclassification between GD and GM. Indeed, the classification performance of the E-nose-LDA-SA could be enhanced if only three groups were considered: RF, GD+GM and GI fruitiness. In this case the E-nose could correctly classify 100% of the oils for training (Figure 2b) and 92% for LOO-CV. In conclusion, the E-nose could be effectively applied as a rapid, cost-effective, and non-invasive tool for olive oil sensory classification.
- Assessing pesticide impact on honeybee gut microbiota: a call for microbial diversity as an environmental risk assessment endpointPublication . Rosa-Fontana, Annelise; Aguado, Daniel; Martín Hernández, Raquel; Higes, Mariano; Henriques, Dora; Pinto, M. AliceA recent roadmap for integrating environmental microbiota into risk assessments under the European Food Safety Authority (EFSA) remit has been published. The honeybee gut microbiota has emerged as a promising avenue to protect bees against stressors. Honeybees exhibit a stable core microbiota, and dysbiosis may serve as an indicator of adverse conditions. We investigated the gut microbiota of newly emerged “Apis mellifera iberiensis” workers exposed to a single concentration of the insecticide flupyradifurone (FPF, 36 ppm). The control groups included pure syrup (negative control, NC) and syrup supplemented with 1% acetone (acetone control, AC). Laboratory trials followed official guidelines (OECD No. 245). The abdomen of each bee was separated from the thorax, and DNA extraction was performed individually. Full-length 16s rRNA amplicon metagenomic was sequenced through PacBio sequel II (HiFi/CCS mode). The Shannon diversity index was used to analyze honeybee gut microbiota composition across experimental groups. Our results revealed a significant increase in bacterial community diversity (Shannon index, P = 0.003) after ten days of chronic exposure to FPF. This effect was more pronounced when compared to the AC group (P = 0.003) than to the NC group (P = 0.03). These findings demonstrate that FPF disrupts the honeybee gut microbiota. This study represents the first characterization of honeybee gut microbiota strictly adhering to OECD guidelines without modifications or adaptations. Furthermore, we have provided new insights into pesticide risk assessment, highlighting an overlooked aspect of bee health assessment. We propose integrating this approach into pesticide risk assessments by using diversity indices as comparative parameters. Specifically, we advocate for the inclusion of honeybee gut microbiota dysbiosis as a sublethal effect in the initial screening phase of risk assessments (laboratory-based assays) and as a key parameter for evaluating pollinator health.
- Assessing the prebiotic potential of xylooligosaccharides produced by one-step fermentation using agro-residuesPublication . Cordeiro, Ana; Fernandes, Andreia; Sousa, Joana; Cardoso, Beatriz; Alves, Joana; Silvério, Sara; Peres, António M.; Rodrigues, Lígia R.; Amorim, CláudiaThe prebiotic effect is a fundamental concept in the fields of nutrition and gut health, referring to the beneficial effects of specific non-digestible dietary components on the gut microbiota, including xylooligosaccharides (XOS). These compounds function as food sources for beneficial gut bacteria, fostering their growth and activity. In this work, in vitro studies were performed to evaluate the prebiotic potential of XOS produced from olive stones (OS) and coffee silver skin (CSS) via a one step fermentation using a recombinant Bacillus subtilis 3610 harbouring the xylanase gene xyn2 from Trichoderma reesei. This potential was compared with a commercially available prebiotic oligofructose (Orafti®, BENEO, Germany). A mixture of human faeces from four healthy donors aged between 24 and 28 years old was used as inoculum. The pH variation and the production of short-chain fatty acids (SCFAs), gases, and ammonia were analysed during the 48 hours fermentations. The prebiotic supplementation resulted in a reduction of the pH value over time, with oligofructose presenting the most significant pH drop at 48 hours (ΔpH=3.65). The addition of prebiotics also significantly increased the production of beneficial SCFAs, with oligofrutose exhibiting a notable increase in the production of lactic and acetic acid production after 48 hours (28.0±0.1 and 28±1 mM, respectively), while OS-XOS and CSS-XOS demonstrated a more prominent rise towards the production of acetic acid (14.8±0.4 and 20.4±0.1 mM, respectively), butyric acid (2.5±0.3 and 3.29±0.04 mM, respectively), and valeric acid (75±1 and 110±14 mM, respectively) at 48 hours. Remarkably, the gas analysis revealed that the addition of OS/CSS-XOS fully suppressed the production of CH4 and increased the CO2 generation after 48 hours (2.6±0.7 and 5.20±0.05 mmol.L-1medium, respectively). These findings strongly suggest that the XOS produced from OS and CSS holds potential prebiotic properties for human health.
- Assessment of farmer's knowledge about fungi and mycotoxin in Southern MozambiquePublication . Bila, João; Macuamule, Custódia; Bombe, Amina; Ribeiro, Maria Isabel; Venâncio, Armando; Afonso, Sandra; Rodrigues, PaulaMycotoxins find their way into the human and animal body through the consumption of mycotoxin contaminated foods, which may result in acute or chronic intoxication. This study aimed to assess knowledge about fungi and mycotoxins among farmers in the provinces of Gaza and Inhamban.e, in southem Mozamhique.Data were collected using quantitative study hased on non-probabilistic questionnaire, covering 180 farmers from Gaza (90) and Inhambane (90) Provinces, from October to November 2022. Data were subjected to descriptive and statistical analysis. The majority of farmers were aged 36 or over (75.0%), were female (75.2%), had no education or only had primary education (80.6%) and had been a farmer for longer 15 years old (64.4%). The level of knowledge about fungi and mycotoxins is mainly explained by the province of residence, followed by the level of education, age and gender, with producers with higher levels of education and of male gender recording a higher level of knowledge. Regarding mycotoxins, most fanners have never heard about these toxic compounds. Likewise, regardless of the province, a significant number of farmers did not know or have never accounted for production losses or income losses due to fungal or mycotoxin contamination.
- Biodiesel production by transesterification using choline hydroxide as catalystPublication . Brito, Paulo; Lima, Renata; Queiroz, Ana; Ribeiro, António E.Biodiesel is a mixture of fatty acid methyl esters (FAMEs) and is a biodegradable and renewable fuel, produced from fat sources mainly composed of triglycerides. The use of ionic liquids (ILs) in biodiesel catalytic production has been studied mainly in the ecological field, as it allows a high recycling efficiency. Choline (2-hydroxyethyl trimethylammonium)-based ILs have received attention due to their biocompatibility characteristics and potential for various industrial applications. Specifically, choline hydroxide (ChOH) represents a promising option. This work's objective is the optimization of the methyl transesterification reaction conditions using commercial and waste sunflower oil (WSO) as raw material and ChOH as a catalyst, assessing the possibility of recovering the catalyst between reaction cycles. Therefore, biodiesel production was carried out on heating plates with temperature control and with magnetic stirring, using methanol reflux. After phase separation, centrifugation was used to enhance biodiesel recovery. Reaction conversion was assessed by acidity drop determination, and the biodiesel FAME content was determined by GC-FID, through a procedure in accordance with EN 14103, using methyl heptadecanoate as the internal standard. IL recovery was carried out by solvent extraction with water-based binary systems, followed by an FTIR analysis of both phases for ChOH detection, and a comparison with initial IL samples. Optimal conversions, determined by acid value (AV) reduction or by biodiesel FAME mass content, were obtained using a 4%wt. catalyst load, oil/methanol molar ratio of 1:8, duration of 1 h, and temperature of 65 °C. The products’ AV for WSO showed a significant reduction relating to the raw material AV (6.14 mgKOH/g). For the reactions with commercial sunflower oil (AV close to 0.20 mgKOH/g), the biodiesel phase AV remained low. ChOH recovery, performed with n-butanol/water and ethyl acetate/water systems, proved to be inefficient under the conditions tested. FTIR analysis showed the presence of ChOH in both liquid–liquid extraction phases.
- Bioinformatics pipeline to evaluate patterns of diversity in detoxification genes in the Western honey bee (Apis mellifera)Publication . Barbosa, Daniela; Li, Fernanda; Bashir, Sana; Lopes, Ana Rita; Yadró Garcia, Carlos A.; Quaresma, Andreia; Rufino, José; Rosa-Fontana, Annelise; Verbinnen, Gilles; de Graaf, Dirk C.; De Smet, Lina; Taliadoros, Demetris; Webster, Matthew; Pinto, M. Alice; Henriques, DoraThe Western honey bee, Apis mellifera, displays significant genetic diversity in detoxification genes, which is pivotal for environmental adaptation and resilience. Herein, we developed a bioinformatics pipeline to investigate patterns of diversity in these genes, focusing on single nucleotide polymorphisms (SNPs) across A. mellifera populations, with variant annotation performed using both snpEff and the Variant Effect Predictor (VEP). Our pipeline integrates GATK, VCFtools, PLINK, bcftools, snpEff, and VEP to process genomic data systematically. Regions of interest were defined in a BED file for variant filtering. Using GATK, SNPs were extracted from a VCF file and conversion to PLINK format for population genetics analyses. Variants were filtered by minor allele frequency (MAF) and population differentiation (FST index) to identify SNPs with considerable. Variants were annotated with snpEff and VEP to predict functional impacts, enabling a comparative analysis of their annotation consistency and depth. Custom scripts were developed to map SNPs to detoxification genes, quantify SNP density, and integrated gene descriptions and lineage data. The resulting data were visualized using a combination of and generate different graphs using ggplot2 and chromoMap for chromossomal maps. Quality control steps were applied through the pipeline ensuring data reliability. Our findings reveal distinct SNP patterns in detoxification genes, highlighting candidate SNPs associated with A. mellifera subspecies-specific adaptations. The comparison of snpEff and VEP annotations provides insights into their strengths and limitations, which can help optimize software selection for genomic studies. This pipeline offers a reproducible framework for studying genetic diversity in A. mellifera that is adaptable to other species, advancing conservation and evolutionary genomics.
- Breakthrough Curves for the Continuous Adsorption of Micropollutants on Activated Carbon/Geopolymer MixturesPublication . Baldo, Arthur P.; Silva, Ana P. F.; Silva, Adriano S.; Díaz de Tuesta, Jose Luis; Marin, Pricila; Peres, José A.; Gomes, Helder T.Breakthrough curves are key for designing continuous flow adsorption systems, typically obtained experimentally or through modeling [1]. In this study, a mixture of activated carbon (AC) and geopolymers (GP) was tested in a continuous treatment system for the removal of model contaminants from waters in a single-component system composed of sulfamethoxazole (SMX), acetaminophen (ACT), or gallic acid (GA). In addition, the adsorbent materials were also tested in a multi-component water matrix to evaluate the interactions between adsorbates and adsorbents. Adsorption tests were conducted using 1g of GP and 0.5g of AC, filled with glass beads. The experiments were carried out at room temperature, considering 100 mg/L for all contaminants and natural pH of the solutions. The flow was set to 1 mL/min and controlled by a peristaltic pump. The breakthrough curves are shown in Figure 1. Based on the experimental data, the useful operating times were calculated, assuming that the effluent concentration should not exceed 5% of the initial concentration [2], yielding 228 minutes for ACT, 46 minutes for SMX, and 53 minutes for GA in the multi-component system. A reduction in time was observed when compared to the single-component system due to faster saturation caused by competition for active sites. The useful adsorption capacities were also calculated, with values of 45.7 mg/g for ACT, 9.2 mg/g for SMX, and 10.7 mg/g for GA in the multicomponent system. The overall results indicated that the column was efficient, as the contaminating compounds were removed up to the system breakthrough. In conclusion, this preliminary study suggests that these materials can be effectively combined and applied in tertiary wastewater treatments, as permeable reactive barriers.
- Comparative evaluation of oranges available for consumption in PortugalPublication . Fajardo, Ana S.; Serra, Valéria; Peres, António M.; Veloso, Ana C.A.; Henriques, Marta H.F.Citrus production in Portugal, particularly in Algarve, plays a significant role in the national and European markets. Oranges from this region are highly regarded for their exceptional quality, due to unique climatic and geographical conditions. Many of these oranges hold the Protected Geographical Indication (PGI) "Citrinos do Algarve" certification, ensuring their authenticity and superior characteristics. While Algarve oranges dominate the market, other varieties also reach consumers, warranting a broader comparative analysis. This study aimed to compare 5 orange varieties, Baía, Dalmau, Navelate, Lane Late, and Salustiana, sourced from different regions, cultivation methods, and market sources. A versatile approach was used, combining physicochemical and sensory analyses, to determine their quality attributes. External properties (appearance, dimensions, and color) were measured along juice parameters such as total solids (TS), density, total soluble solids (SS) (°Brix), titratable acidity (TA), SS/TA ratio, and color. Sensory perception was assessed by an untrained panel evaluating aspects like appearance, taste, sweetness, and acidity. Additionally, an electronic tongue (E-tongue) was employed to detect and analyze chemical patterns associated with sweetness perception, ensuring precise and consistent results. Results showed all varieties except Dalmau had over 70% sensory acceptance, with scores exceeding 4.2 on a 7-point scale. Lane Late variety was the most favored among consumers, excelling in both external attributes and taste. Lane Late physicochemical analysis supported these preferences: pH of 3.53 ± 0.12, TA of 0.50 ± 0.12 g citric acid/100 g, SS of 12.14 ± 0.66 °Brix, and the highest SS/TA ratio of 22.99 ± 3.27, reinforcing its perceived sweetness. E-tongue analysis successfully distinguished flavor profiles. A classification model using 40 lipid sensor membranes achieved a 94.4% correct classification rate. The linear discriminant analysis (LDA) model, enhanced by the simulated annealing (SA) variable selection algorithm, efficiently separated the 5 orange cultivars. Two discriminant functions (DF1 accounting for 99.8% variability and DF2 for 0.10%) effectively differentiated the samples. The E-tongue's ability to detect and classify sweet and acidic sensations contributed to its strong predictive performance. This integrated analytical approach provided a comprehensive comparison of the sweetness profiles of oranges available in Portugal, blending objective and subjective evaluations. The study reaffirmed the superior qualities of Algarve oranges, with the PGI-certified Lane Late standing out. With an average diameter of 87.41 ± 3.76 mm, second only to Baía (92.14 ± 5.40 mm), and an intermediate peel thickness of 5.27 ± 0.58 mm, Lane Late demonstrated notable physical attributes. Its TS content (12.01 ± 0.72%) was close to Navelate (12.62 ± 1.49%), while it had the lowest ash content (0.38 ± 1.13%), reinforcing its distinctiveness.
- Comprehensive analysis of oranges available for consumption in Portugal: A comparative studyPublication . Fajardo, Ana S.; Serra, Valéria; Peres, António M.; Veloso, Ana C.A.; Henriques, Marta H.F.Citrus production in Portugal's Algarve region is a major economic activity, contributing to both local and national markets with exports primarily to Europe.