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- Advice on how to start beekeeping, memories with bees and the uses of honey: results of an online questionnaire with European beekeepersPublication . Perichon, Samuel; Adamchuk, Leonora; Biber, Lejla; Božič, Janko; Chlebo, Róbert; Filipi, Janja; Leidenberger, Sonja; Mavrofridis, Georgios; Özgör, Erkay; Pocol, Cristina Bianca; Porporato, Marco; Rodríguez-Flores, María Shantal; Vilas-Boas, Miguel; Zacepins, AleksejsBeekeeping is a demanding activity that requires both particular human qualities from those who practise it and an environment that is favourable to bees. This is why the interviewed beekeepers advise to take time to think before starting to keep the bees, to find a mentor, and to always have the desire to get to know the bees even if this is not enough for success. In Northern and Western Europe, beekeepers consider patience and calmness as essential qualities to be a “good beekeeper”, while in Southern Europe, passion for bees is the main driver of success. The reasons for abandonment or failure also include human and environmental factors. To better understand the relationship between beekeepers and their bees, interviewees were encouraged to share their best memories. With years of experience, contemplative memories fade into memorable situations in which bee practices are described, and then in turn, they fade into moments of sociability. Honey harvesting plays an important role. Productive considerations are more expressed in Southern Europe, where it is an exceptional harvest that marks on people’s minds, more than the first honey harvest. The strong connection to their bees and the territory they live in is also expressed in the choice of their favourite honey. Many beekeepers mentioned the local honey production, which they are proud to produce with their own bees, even though they sometimes became criticized for a too high price for this local product. © 2024 International Bee Research Association.
- 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.
- Application of encapsulated tomato by-product extract as a colorant in mascarponePublication . Molina, Adriana K.; Leichtweis, Maria Gabriela; Machado, Manuela; Silva, Sara; Pintado, Manuela; Barreira, João C.M.; Dias, Maria Inês; Prieto Lage, Miguel A.; Barros, Lillian; Pereira, CarlaAgricultural by-products, often underutilized, represent a significant opportunity to obtain sustainable value-added products to develop new functional foods. In this context, the (typically discarded) aerial parts of tomato plants (Solanum lycopersicum L. var. cerasiforme), were studied as new sources of natural bioactive compounds, focusing the extraction and encapsulation (using spray drying with maltodextrin as the encapsulant) of chlorophylls. The encapsulated natural colorant was subsequently incorporated into mascarpone cheese to assess its stability and effect on nutritional and antioxidant properties. The obtained extracts were mainly characterized by high concentrations of bioactive compounds, particularly quercetin-3-O-deoxyhexoside (14.4 ± 0.3 mg/g). The colorant imparted a greener hue to the mascarpone (a* –6.0 ± 0.05 vs. –1.7 ± 0.05 in control, p < 0.001) and maintained antioxidant activity (OxHLIA EC₅₀ = 1440 ± 72 to 1166 ± 66 μg/mL, p = 0.030) without altering the nutritional profile (p > 0.05). Moreover, no cytotoxicity was observed, and the fatty acid composition remained unchanged. Accordingly, the studied encapsulated colorant might be used as a functional ingredient in other dairy products, imparting an attractive color while preserving nutritional integrity. In conclusion, the incorporation of TAPC is an effective strategy to enrich dairy products with natural pigments and bioactive compounds without compromising quality. Future research is needed to explore its application in other food matrices and assess long-term storage effects.
- 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.
- Assessment of farmer’s knowledge and attitudes toward fungi and mycotoxin contamination in staple crops in Southern MozambiquePublication . Bila, João; Macuamule, Custódia; Bombe, Amina; Ribeiro, Maria Isabel; Venâncio, Armando; Afonso, Sandra; Rodrigues, PaulaIn Mozambique, 80% of the population directly depends on agriculture as a source of food and income. However, some of the most produced food crops, such as maize, rice and peanuts, are easily contaminated by fungi and mycotoxins. The naturally high prevalence of mycotoxins can be aggravated by the high vulnerability and lack of knowledge of the farmers. The aims of this study were to assess the knowledge and perceptions of small-size and medium-size farmers in the provinces of Inhambane and Gaza, southern Mozambique, regarding awareness of fungi and mycotoxin contamination of food crops, losses of production and income, and the causes and consequences of this contamination. A survey was conducted with 180 farmers in the two provinces. A multiple linear regression model was used to correlate the level of knowledge with the sociodemographic characteristics of the studied population. Results The results showed that 97.8% of the farmers have an insufficient level of knowledge about fungi and mycotoxins contamination of food crops. While 17.8% showed sufficient or good knowledge of the conditions that promote fungal contamination, only 3.9% knew what measures to apply to mitigate their occurrence. The level of knowledge was lower for the Inhambane farmers. According to the estimated model, province, gender, age (>45 years old), primary and secondary (1st cycle) education, another source of income other than agriculture and experience as a farmer (>10 years) are statistically significant predictors of the level of knowledge of the Mozambican farmers analyzed. These findings highlight the urgent need of tailored interventions to promote good agricultural and storage practices that allow the mitigation of mycotoxin contamination of food.
- Assessment of Trioza erytreae microbiome and mitochondrial genome variability by integrated high-throughput sequencing approachPublication . Chiumenti, Michela; Nicoloso, Vittorio; Fereres, Alberto; Pereira, José Alberto; Maree, Hans J.; Bester, Rachelle; Reynaud, Bernard; Delatte, Hélène; Peña, Leandro; Pallás, Vicente; Serra, Pedro; Navarro, Beatriz; Serio, Francesco DiHuanglongbing, a devastating citrus disease, is associated with 'Candidatus Liberibacter asiaticus', 'Ca. L. africanus' or 'Ca L. americanus', bacteria transmitted by the psylloids Diaphorina citri and Trioza erytreae. Using a DNA-Seq and metabarcode sequencing integrated approach, the first catalogue of endosymbionts associated with T. erytreae from the Iberian Peninsula, South Africa and African Islands, was generated. The almost complete genome of two new bacteria, one facultative and one obligate, tentatively named Asaia-like endosymbiont of T. erytreae and Sodalis-like endosymbiont of T. erytreae, respectively, was assembled and annotated. The complete mitochondrial genomes of T. erytreae from the geographical areas studied were also assembled and phylogenetic analyses were performed, suggesting that T. erytreae populations currently present in the Iberian Peninsula and specimens analyzed from South Africa may have originated from a common ancestor. Similar results were obtained when the genetic distances between Sodalis-like endosymbiont of T. erytreae were taken into consideration, thus supporting the symbiont-host codivergence which suggests that this bacterium is approaching to an obligate status. Finally, a new genetic marker of T. erytreae, an insertion in the mitochondrial tRNA-Ser gene, was identified only in some European samples, showing for the first time the existence of two mixed subpopulations of T. erytreae. The integrated DNA-Seq and metabarcode sequencing approach used in this study, besides generating a catalogue of T. erytreae endosymbionts, provided novel data on the sequence variability of bacterial and insect mitochondrial genomes from different geographic areas, highlighting the possible original sources of currently spreading T. erytreae populations may be more complex than previously reported.
- 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.
- Bioactive metabolites from algae: occurrence, extraction techniques, functional properties, food applications and therapeutic prospectsPublication . Pereira, Cláudia S.G.P.; Carpena, M.; Barreira, João C.M.; Silva, Cristiana F.C.; Prieto Lage, Miguel A.; Oliveira, BeatrizThis review critically summarizes the major algal-derived bioactive compounds, including phenolics compounds (e.g., phlorotannins, bromophenols, phenolic acids), sterols (e.g., fucosterol, beta-sitosterol), and polysaccharides (e. g., carrageenan, fucoidan, ulvan), as well as key pigments such as astaxanthin, beta-carotene, lutein, violaxanthin, zeaxanthin, and chlorophylls. In addition, state-of-the-art extraction technologies including supercritical fluid extraction, pressurized liquid extraction, ultrasound assisted, microwave assisted, surfactant assisted, and enzyme assisted extractions, and carbon dioxide-expanded liquids) were systematically compared considering key parameters such as yield, versatility, compound selectivity, extract quality, cost-effectiveness, technical complexity, operational conditions, and sustainability principles. Based on this comparative evaluation, ultrasound assisted extraction emerged as the most versatile among advanced technologies, same as enzyme assisted extraction within ulterior techniques. The potential incorporation of these bioactive compounds into novel algaebased food systems, either as functional ingredients or texturizing agents, was also discussed. Moreover, their relevance in alternative therapeutic applications was also highlighted, with a proof-of-concept focused on the antiviral potential of sulfated polysaccharides (carrageenan, fucoidan, and ulvan). Overall, algae-based ingredients were found to be promising biofunctional agents for innovative food products, nutraceutical formulations, and therapeutic alternatives.
- Carob and pomegranate extracts enhance plant defence mechanisms against olive anthracnose through antioxidant activity and phenolic compounds productionPublication . Antón-Domínguez, Begoña I.; Sánchez-Pereira, Luiza; Lamas, Sandra; Rodrigues, Nuno; Baptista, Paula; Agustí-Brisach, CarlosOlive anthracnose (OA), caused by Colletotrichum species, is one of the most economically damaging disease in olive sector. This study was focused to identify sustainable control alternatives by evaluating the effect of pomegranate and carob extracts against OA and elucidating their mode of action. In vitro assays on mycelial growth, conidial production, and germination, and appressoria formation of C. godetiae and C. nymphaeae were performed. Neither extract significantly inhibited mycelial growth, while both reduced reproductive structures formation of the pathogens. Bioassays on detached and attached olive fruits in plants of cv. Arbequina, treated with extracts and/or inoculated with C. godetiae were conducted to evaluate the effect of the extracts on disease progression. Despite of the limited effect of the extracts on reducing disease progression in detached fruit, curative applications were more effective than preventive ones. Carob extract was more effective than pomegranate extract in reducing the disease incidence progression in attached fruit in planta. The resistance-inducing effect of plant extracts was evaluated by quantifying of H2O2 and phenolic compounds production in olive leaves at 0, 3, 7 and 24 h after inoculation with C. godetiae. Both extracts increased these parameters in the inoculated plants, with carob extract triggering an earlier activation and promoting a greater diversity of phenolic compounds accumulation. These findings reveals that these two extracts act as resistance inducers through different effective defence pathways, with carob extract standing out as a promising bioprotector against OA. These results open new possibilities for environmentally friendly management of OA using plant extracts.