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- 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 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.
- Benchmarking lamp primer design platforms for pyrethroid resistance SNP detection in varroa destructorPublication . Bejaoui, Mohamed; Costa, Maíra; Pinto, M. Alice; Henriques, DoraVarroa destructor, an ectoparasitic mite of Apis mellifera, is a driver of colony and pollination service declines. Widely used pyrethroid acaricides targeting the mite voltage-gated sodium channel (VGSC) have selected for resistance mutations at codons 918 and 925 in Domain II. Genotyping 100 mites from 35 apiaries at the locus revealed ~40% pyrethroid-resistant haplotypes, with the double-resistant M918L/L925V variant at 43%, establishing the Portuguese baseline. PCR-based assays at this locus are robust but laboratory-bound, whereas loop-mediated amplification (LAMP) offers an isothermal, rapid, low-cost alternative for field resistance surveillance. Using this baseline, we benchmarked mutation - focused LAMP primer sets with PrimerExplorer V5, NEB LAMP Designer, and LAMP Designer v1.16 under shared, explicitly defined thermodynamic constraints. We constrained outer primers and inner-primer Tm to 55–70 °C, loops to ≈62–65 °C, GC to 40–70%, F2–B2 span to 120–180 bp, and filtered candidates using ΔG thresholds for hairpins and dimers. PrimerExplorer V5 provided flexible control over primer geometry around codons and loop placement, but required a loop-primer step and external specificity checks. NEB LAMP Designer rapidly generated optimised sets with conservative end-stability and self-dimer filtering, although limited parameter tuning and absence of BLAST restricted mutation-centred optimization. LAMP Designer v1.16 delivered an exhaustive thermodynamic assessment of hairpins, self-dimers, and cross-dimers together with BLAST-supported specificity and evaluation, at the cost of a steeper learning curve and reliance on proprietary software. In NEB LAMP Designer and LAMP Designer v1.16, targeted codons were embedded in distinct inner-primer segments (918 in F2, F1c; 925 in F1c, B1c, respectively), whereas PrimerExplorer V5 was used to design assays on codon 925 alone, given that M918L never occurred without L925V. Collectively, these comparisons show how software architecture and Tm/ΔG criteria shape LAMP primer solutions for resistance SNPs and inform the design of field-deployable diagnostics for evolutionary surveillance.
- 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.
- Desarrollo de modelos predictivos para la selección de genotipos de aceituna de mesa menos susceptibles a Bactrocera oleae (Rossi)Publication . González-Fernández, Antonio; Rallo, Pilar; Peres, António M.; Pereira, José Alberto; Morales-Sillero, AnaLa mosca del olivo (Bactrocera oleae Rossi), considerada como la plaga más devastadora en los olivares, causa importantes pérdidas en la producción y en la calidad de la aceituna de mesa y del aceite de oliva a nivel mundial. Esto conlleva a que se le otorgue atención a la tolerancia a esta plaga en los programas de mejora del olivo durante el proceso de selección de genotipos. Son múltiples los factores que influyen en la preferencia de la mosca del olivo por ciertas variedades, entre otros los parámetros físicos y químicos del fruto, que deben interpretarse de forma colectiva. Los objetivos de este trabajo han sido identificar las diferencias de susceptibilidad a B. oleae en una selección de genotipos y desarrollar modelos predictivos basados en rasgos físicos y químicos del fruto, capaces de predecir el comportamiento de la mosca. Para ello se han empleado frutos de cuatro genotipos: dos selecciones avanzadas del programa de mejora genética de aceituna de mesa de la Universidad de Sevilla, y dos variedades tradicionales muy apreciadas para el aderezo (US 06-1388; US-06-194; ‘Hojiblanca’; ‘Kalamon’), todos procedentes de un ensayo de material vegetal localizado en Morón de la Frontera (Sevilla). En condiciones controladas de laboratorio, se ha estudiado, por un lado, la preferencia de la mosca mediante bioensayos de oviposición y, por otro lado, se han desarrollado modelos predictivos basados en parámetros como el peso, tamaño, color y textura del fruto, además de los contenidos en compuestos fenólicos.
- Different approaches to Olive Pomace valorisationPublication . Cordeiro, Ana; Lamas, Sandra; Rodrigues, Ana; Cadavez, Vasco; Silvério, Sara; Peres, António M.; Rodrigues, Lígia R.; Amorim, CláudiaOlive pomace (OP) is a substantial by-product, rich in lignocellulosic materials and water, obtained following the extraction of olive oil. It comprises the following components: olive skins, pulp, seeds, and stones. As global olive oil production continues to rise, the management and utilisation of OP have become increasingly important from both an economic and an environmental perspective [1]. In this study, the OP was chemically characterized and its potential as a source of value-added compounds was evaluated. For this purpose, several extraction studies were preformed, namely lipid extraction using petroleum ether, phenolic compound extraction with ethanol, and free sugar extraction with water using a Soxhlet system (Figure 1). These extractions allowed to recover 11.1 ± 0.3 % (w/w) of lipids, different types of phenolic compounds and oligosaccharides. Phenolic compounds are widely recognized for their health-promoting properties (e.g. antioxidant, anti-inflammatory, cancer prevention or antimicrobial activity) and have been the subject of extensive research. Oligosaccharides (OS) are short chains of monosaccharide units linked by glycosidic bonds. The ingestion of these compounds has been demonstrated to confer a multitude of salutary effects, predominantly due to their function as prebiotics and their capacity to regulate gut health and influence overall metabolic processes. Enzymatic hydrolysis was also performed using commercial xylanase from Trichoderma reesei, under optimal conditions (pH 4.5, 40°C, 150 rpm) to evaluate the potential of OP to produce xylo-based oligosaccharides [2]. The hydrolysis process was monitored through Thin Layer Chromatography (TLC), High-Performance Liquid Chromatography (HPLC) using different columns, and the Dinitrosalicylic acid (DNS) method (Figure 2). The results indicated the production of xylo-based oligosaccharides, validating the proposed bioprocess as a viable method for XOS production from OP. This approach not only provides value-added to OP but also aligns with sustainable waste management practices.
- Discriminação de azeites ‘Verdeal Transmontana’ de acordo com a idade das oliveiras recorrendo à espectroscopia no infravermelho por transformada de FourierPublication . Lamas, Sandra; Baptista, Paula; Santamaria-Echart, Arantzazu; Barreiro, Filomena; Peres, António M.; Pereira, José Alberto; Rodrigues, Nuno; Baptista, PaulaO efeito da idade das oliveiras na qualidade do azeite é ainda pouco conhecido. Nesse sentido, o presente trabalho teve como objetivo avaliar a influência da idade da oliveira na qualidade e composição dos azeites, usando a cv. Verdeal Transmontana como modelo. Para tal foram selecionadas 15 oliveiras jovens (< 20 anos), 15 com idade intermédia (entre 20 e 50 anos) e 15 centenárias (> 100 anos), de cujas azeitonas foram extraídos azeites. Nos azeites avaliaram-se os parâmetros de qualidade (acidez, índice de peróxidos, coeficiente de extinção específica no ultravioleta, avaliação sensorial), atividade antioxidante e estabelecidos os espetros no infravermelho por transformada de Fourier (FTIR). Todos os azeites obtidos podem ser classificados como azeite virgem extra. A idade da planta influenciou alguns parâmetros, sem que tenha uma tendência linear. Os azeites extraídos de oliveiras com idade intermédia, apresentaram valores superiores de atividade antioxidante e teor em fenóis totais. A aplicação da análise discriminante linear (LDA), conjuntamente com o algoritmo metaheurístico de seleção de variáveis (SA), aos espetros de FTIR, permitiram estabelecer um modelo FTIR-LDA-SA, com base nas transmitâncias (%) registadas para 10 números de onda selecionados (2860, 2856, 2783, 1329, 1094, 918, 739, 723, 683, e 615 cm-¹). O modelo apresentou sensibilidade e especificidade de 100%, permitindo identificar corretamente as idades de todas as oliveiras estudadas, com base nos espectros FTIR, de acordo com a validação cruzada k-fold. O sucesso alcançado na previsão permite concluir que a técnica FTIR pode ser usada como uma ferramenta não invasiva/não-destrutiva de identificação da idade das oliveiras com base nos espectros de transmitância dos azeites extraídos.
- Distribution of pyrethroid and amitraz resistance in Varroa destructor mites from honey bee (Apis mellifera) colonies in PortugalPublication . Costa, Maíra; Yadró Garcia, Carlos A.; Rodrigues, Cláudia; Lopes, Ana Rita; Pérez-Pérez, Antonio; Martín Hernández, Raquel; Higes, Mariano; Pinto, M. Alice; Henriques, DoraHoney bees (Apis mellifera) are widely used for pollination and honey production worldwide. However, they face a serious threat from the ectoparasitic mite Varroa destructor. This parasite causes varroosis and transmits multiple viruses, such as the Deformed wing virus (DWVV), compromising the health and survival of colonies. While there are many acaricides commercially available to treat colonies against varroa, the recurrent use of certain molecules is leading to the development of varroa resistance. In Portugal, two classes of synthetic compounds are currently used: pyrethroids (fluvalinate and flumethrin) and formamidines (amitraz). However, the excessive and repeated use of acaricides has led to the development of resistance in mite populations. Pyrethroid resistance is linked to mutations in the Voltage-Gated Sodium Channel (VGSC) gene, with amino acid substitutions, at position 925 and 918. Amitraz resistance is associated with mutations in the Octopamine-like β-adrenergic receptor (Octβ2R). The amino acid changes include asparagine (N) to serine (S) at position 87 (N87S), observed in France; tyrosine (Y) to histidine (H) at position 215 (Y215H), in the USA; phenylalanine (F) to leucine (L) at position 290 (F290L), in Spain; and tyrosine (Y) to phenylalanine (F) at position 337 (Y337F), in Turkey. The distribution of resistance alleles has not yet been explored in Portugal. To fill this data gap, Varroa mites from honey bee colonies across various regions of the country were analyzed through DNA extraction, PCR with specific primers, and Sanger sequencing. The results confirmed the of widespread distribution of resistance alleles to pyrethroids at positions 925 and 918, like those observed in Spain. In contrast, preliminary results did not detect any amitraz resistance alleles, although the small sample size warrants caution in interpretation. This study represents an important step in understanding V. destructor resistance to chemical treatments in Portugal, providing essential data for monitoring acaricide resistance and supporting the development of more effective management strategies to help beekeepers better protect their colonies.