The association of milk and dairy consumption with iodine status in pregnant women in Oporto region.

The role of milk and dairy products in supplying iodine to pregnant women is unknown in Portugal. The aim of this study was to evaluate the association between milk and dairy product consumption and the iodine status of pregnant women in the IoMum cohort of the Oporto region. Pregnant women were recruited between 10 and 13 weeks of gestation, when they provided a spot urine sample and information on lifestyle and intake of iodine-rich foods. Urinary iodine concentration (UIC) was determined by inductively coupled plasma MS. A total of 468 pregnant women (269 iodine supplement users and 199 non-supplement users) were considered eligible for analysis. Milk (but not yogurt or cheese) intake was positively associated with UIC, in the whole population (P = 0·02) and in the non-supplement users (P = 0·002), but not in the supplement users (P = 0·29). In non-supplement users, adjusted multinomial logistic regression analysis showed that milk consumption <3 times/month was associated with a five times increased risk of having UIC < 50 µg/l when compared with milk consumption ≥2 times/d (OR 5·4; 95 % CI 1·55, 18·78; P = 0·008). The highest UIC was observed in supplement users who reported consuming milk once per d (160 µg/l). Milk, but not yogurt or cheese, was positively associated with iodine status of pregnant women. Despite the observed positive association, daily milk consumption may not be sufficient to ensure adequate iodine intake in this population.

Iodine knowledge is associated with iodine status in Portuguese pregnant women: results from the IoMum cohort study.

Lack of knowledge about iodine has been suggested as a risk factor for iodine deficiency in pregnant women, but no studies have addressed this issue in Portugal. So, the aim of this study was to investigate iodine knowledge among Portuguese pregnant women and its association with iodine status. IoMum, a prospective observational study, included 485 pregnant women recruited at Centro Hospitalar e Universitário de S. João, Porto, between the 10th and 13th gestational weeks. Partial scores for knowledge on iodine importance, on iodine food sources or on iodised salt were obtained through the application of a structured questionnaire. Then, a total iodine knowledge score was calculated and grouped into low, medium and high knowledge categories. Urinary iodine concentration (UIC) was measured in spot urine samples by inductively coupled plasma MS. Of the pregnant women, 54 % correctly recognised iodine as important to neurocognitive development, 32 % were unable to identify any iodine-rich food and 71 % presented lack of knowledge regarding iodised salt. Of the women, 61 % had a medium total score of iodine knowledge. Knowledge on iodine importance during pregnancy was positively associated with iodine supplementation and also with UIC. Nevertheless, median UIC in women who correctly recognised the importance of iodine was below the cut-off for adequacy in pregnancy (150 µg/l). In conclusion, knowledge on iodine importance is positively associated with iodine status. Despite this, recognising iodine importance during pregnancy may not be sufficient to ensure iodine adequacy. Literacy-promoting actions are urgently needed to improve iodine status in pregnancy.

Lead Levels in Non-Occupationally Exposed Women with Preeclampsia.

There are many controversies regarding the relationship between lead exposure andcomplications in pregnancy. Preeclampsia (PE) is a maternal hypertensive disorder which is one of the main causes of maternal and foetal mortality. The aim of our study was to assess blood lead level (BLL) in Polish women with PE (PE group, n = 66) compared with healthy, non-pregnant women (CNP group, n = 40) and healthy pregnant women (CP group, n = 40). BLL was determined by inductively coupled plasma mass spectrometry (ICP-MS). The systolic blood pressure (SBP), diastolic blood pressure (DBP) and BLL in the CP group were significantly lower than in the PE group (p < 0.001). Logistic regression analyses of BLL showed a significant positive relationship with the presence of PE. Furthermore, both the SBP and DBP values were positively associated with BLL. This study indicates that preeclamptic women tend to present with significantly higher BLL compared to healthy pregnant women. There were no differences in the BLL between the CP and CNP groups.

Anomalous origin of right pulmonary artery: diagnosis, treatment, and follow-up in an adult patient.

Anomalous origin of a branch pulmonary artery from the aorta is a rare malformation, accounting for 0.12% of all congenital heart defects. We present the case of a 43-year-old man with an anomalous origin of the right pulmonary artery (AORPA) from the ascending aorta. Reimplantation of the right pulmonary artery was carried out successfully, with favourable evolution in the medium-term follow-up. It is the first described case that receives corrective treatment in adulthood with a favourable evolution.

Elemental impurities in lipsticks: Results from a survey of the Portuguese and Brazilian markets.

For safety reasons, European regulations prohibit the use of a long list of metal(loid)s as ingredients of cosmetic products. However, their presence as impurities in finished products is virtually unavoidable, even under GMP conditions. This study aimed at determining the elemental profile of lipsticks available in the Portuguese and Brazilian markets. A total of 96 lipsticks were purchased in Brazil (n = 53; 9 brands) and Portugal (n = 43; 7 brands) and the content of 44 elements was determined. Results ranged from <1 µg/g to several tens of µg/g (e.g., Sn, Mn, Zn). Significant differences were found between Portuguese and Brazilian products for several elements, particularly for Pb. For the elements of major toxicological concern (Pb, Cd, As, Sb, Hg), mean values were always below the current limits set by the German competent authority. However, a significant percentage of exceedances were observed for Pb (24%) and Cd (21%). A safety assessment was carried out for the toxicologically relevant elements. Results showed that, except for Pb, the systemic exposure resulting from lipstick use represents less than 0.2% (ca. 3% for Pb) of the respective permitted daily exposure even in the worst-case scenario (i.e., ingestion of the total amount of product applied).

Effects of Chrysosporum (Aphanizomenon) ovalisporum extracts containing cylindrospermopsin on growth, photosynthetic capacity, and mineral content of carrots (Daucus carota).

Natural toxins produced by freshwater cyanobacteria, such as cylindrospermopsin, have been regarded as an emergent environmental threat. Despite the risks for food safety, the impact of these water contaminants in agriculture is not yet fully understood. Carrots (Daucus carota) are root vegetables, extensively consumed worldwide with great importance for human nourishment and economy. It is, therefore, important to evaluate the possible effects of using water contaminated with cyanotoxins on carrot cultivation. The aim of this work was to investigate cylindrospermopsin effects on D. carota grown in soil and irrigated for 30 days, with a Chrysosporum ovalisporum extract containing environmentally relevant concentrations of cylindrospermopsin (10 and 50 µg/L). The parameters evaluated were plant growth, photosynthetic capacity, and nutritional value (mineral content) in roots of carrots, as these are the edible parts of this plant crop. The results show that, exposure to cylindrospermopsin did not have a clear negative effect on growth or photosynthesis of D. carota, even leading to an increase of both parameters. However, alterations in mineral contents were detected after exposure to crude extracts of C. ovalisporum containing cylindrospermopsin. A general decline was observed for most minerals (Ca, Mg, Na, Fe, Mn, Zn, Mo, and P), although an increase was shown in the case of K and Cu, pointing to a possible interference of the cyanobacterial extract in mineral uptake. This study is the first to evaluate the effects of C. ovalisporum extracts on a root vegetable, however, more research is necessary to understand the effects of this toxin in environmentally relevant scenarios.

Trace Elements in Ambient Air at Porto Metropolitan Area-Checking for Compliance with New European Union (EU) Air Quality Standards.

Because of the scientific evidence showing that arsenic (As), cadmium (Cd), and nickel (Ni) are human genotoxic carcinogens, the European Union (EU) recently set target values for metal concentration in ambient air (As: 6 ng/m(3), Cd: 5 ng/m(3), Ni: 20 ng/m(3)). The aim of our study was to determine the concentration levels of these trace elements in Porto Metropolitan Area (PMA) in order to assess whether compliance was occurring with these new EU air quality standards. Fine (PM2.5) and inhalable (PM10) air particles were collected from October 2011 to July 2012 at two different (urban and suburban) locations in PMA. Samples were analyzed for trace elements content by inductively coupled plasma-mass spectrometry (ICP-MS). The study focused on determination of differences in trace elements concentration between the two sites, and between PM2.5 and PM10, in order to gather information regarding emission sources. Except for chromium (Cr), the concentration of all trace elements was higher at the urban site. However, results for As, Cd, Ni, and lead (Pb) were well below the EU limit/target values (As: 1.49 ± 0.71 ng/m(3); Cd: 1.67 ± 0.92 ng/m(3); Ni: 3.43 ± 3.23 ng/m(3); Pb: 17.1 ± 10.1 ng/m(3)) in the worst-case scenario. Arsenic, Cd, Ni, Pb, antimony (Sb), selenium (Se), vanadium (V), and zinc (Zn) were predominantly associated to PM2.5, indicating that anthropogenic sources such as industry and road traffic are the main source of these elements. High enrichment factors (EF > 100) were obtained for As, Cd, Pb, Sb, Se, and Zn, further confirming their anthropogenic origin.

Effects of microcystin-LR, cylindrospermopsin and a microcystin-LR/cylindrospermopsin mixture on growth, oxidative stress and mineral content in lettuce plants (Lactuca sativa L.).

Toxic cyanobacterial blooms are documented worldwide as an emerging environmental concern. Recent studies support the hypothesis that microcystin-LR (MC-LR) and cylindrospermopsin (CYN) produce toxic effects in crop plants. Lettuce (Lactuca sativa L.) is an important commercial leafy vegetable that supplies essential elements for human nutrition; thus, the study of its sensitivity to MC-LR, CYN and a MC-LR/CYN mixture is of major relevance. This study aimed to assess the effects of environmentally relevant concentrations (1, 10 and 100 µg/L) of MC-LR, CYN and a MC-LR/CYN mixture on growth, antioxidant defense system and mineral content in lettuce plants. In almost all treatments, an increase in root fresh weight was obtained; however, the fresh weight of leaves was significantly decreased in plants exposed to 100 µg/L concentrations of each toxin and the toxin mixture. Overall, GST activity was significantly increased in roots, contrary to GPx activity, which decreased in roots and leaves. The mineral content in lettuce leaves changed due to its exposure to cyanotoxins; in general, the mineral content decreased with MC-LR and increased with CYN, and apparently these effects are time and concentration-dependent. The effects of the MC-LR/CYN mixture were almost always similar to the single cyanotoxins, although MC-LR seems to be more toxic than CYN. Our results suggest that lettuce plants in non-early stages of development are able to cope with lower concentrations of MC-LR, CYN and the MC-LR/CYN mixture; however, higher concentrations (100 µg/L) can affect both lettuce yield and nutritional quality.

Assessment of metal(loid)s phytoavailability in intensive agricultural soils by the application of single extractions to rhizosphere soil.

The influence of soil properties on the phytoavailability of metal(loid)s in a soil-plant system was evaluated. The content of extractable metal(loid)s obtained by using different extraction methods was also compared. To perform this study, a test plant (Lactuca sativa) and rhizosphere soil were sampled at 5 different time points (2, 4, 6, 8 and 10 weeks of plant growth). Four extraction methods (Mehlich 3, DTPA, NH4NO3 and CaCl2) were used. Significant positive correlations between the soil extractable content and lettuce shoot content were obtained for several metal(loid)s. The extraction with NH4NO3 showed the higher number of strong positive correlations indicating the suitability of this method to estimate metal(loid)s phytoavailability. The soil CEC, OM, pH, texture and oxides content significantly influenced the distribution of metal(loid)s between the phytoavailable and non-phytoavailable fractions. A reliable prediction model for Cr, V, Ni, As, Pb, Co, Cd, and Sb phytoavailability was obtained considering the amount of metal(loid) extracted by the NH4NO3 method and the main soil properties. This work shows that the analysis of rhizosphere soil by single extractions methods is a reliable approach to estimate metal(loid)s phytoavailability.

Technological and textural properties of gluten-free quinoa-based pasta (Chenopodium quinoa Wild).

Quinoa (Chenopodium quinoa Willd.) is an Andean grain with a perfect nutritional composition that, by diversifying its transformation, becomes an attractive alternative for consumers looking for a high-quality, healthy diet with a source of vegetable proteins. The objective of this work was to elaborate and evaluate the technological and textural properties of quinoa paste in its entirety through the Star-Shaped Composite Central Design (CCD) of 5 process variables: Water Temperature (°C), Water Quality (ml), Mixing Time (min), Drying Temperature (°C), and Drying Time (min), with 5 levels each. At the same time, the yield and good cooking quality were studied to optimize the process. In the model of the equation for the cooking time response, a negative and significant influence of drying temperature was shown. On the other hand, for cooking loss, dough gain, a* and b* values, and texture had high values if the drying time was increased. On the contrary, the L-value decreases, which is positively significant. Meanwhile, the swelling index was only significantly positive within the technological properties. In addition, it was found that the optimal conditions for producing quality pasta were 25 °C: 1150 ml: 30 min: 70 °C and 80 min, respectively, with a desirability of 0.883. When the pasta was prepared with quinoa, the cooking time was 7 min, the cooking loss was 2.46 g/g, the mass gain was 23.6 g/g, the cooking yield was 7.99%, the swelling index was 2.9%, water absorption was 135%, and protein was 12.71 g and 0. 21 Pa in texture, these results being consistent with cited research. Likewise, the whiteness was 51.97 for the values a* 2.41 and b* 12.45; all this analysis is reflected in the final yield of the process at 78%. In conclusion, the results indicated that, by optimizing the conditions in the production of gluten-free quinoa pasta, it is possible to obtain a gluten-free product with high added value, excellent cooking quality, adequate technological properties, texture, and color acceptable to the consumer.

An eco-friendly approach for analysing sugars, minerals, and colour in brown sugar using digital image processing and machine learning.

Brown sugar is a natural sweetener obtained by thermal processing, with interesting nutritional characteristics. However, it has significant sensory variability, which directly affects product quality and consumer choice. Therefore, developing rapid methods for its quality control is desirable. This work proposes a fast, environmentally friendly, and accurate method for the simultaneous analysis of sucrose, reducing sugars, minerals and ICUMSA colour in brown sugar, using an innovative strategy that combines digital image processing acquired by smartphone cell with machine learning. Data extracted from the digital images, as well as experimentally determined contents of the physicochemical characteristics and elemental profile were the variables adopted for building predictive regression models by applying the kNN algorithm. The models achieved the highest predictive capacity for the Ca, ICUMSA colour, Fe and Zn, with coefficients of determination (R2) ≥ 92.33 %. Lower R2 values were observed for sucrose (81.16 %), reducing sugars (85.67 %), Mn (83.36 %) and Mg (86.97 %). Low data dispersion was found for all the predictive models generated (RMSE < 0.235). The AGREE Metric assessed the green profile and determined that the proposed approach is superior in relation to conventional methods because it avoids the use of solvents and toxic reagents, consumes minimal energy, produces no toxic waste, and is safer for analysts. The combination of digital image processing (DIP) and the kNN algorithm provides a fast, non-invasive and sustainable analytical approach. It streamlines and improves quality control of brown sugar, enabling the production of sweeteners that meet consumer demands and industry standards.

High-Value Compounds in Papaya By-Products (Carica papaya L. var. Formosa and Aliança): Potential Sustainable Use and Exploitation.

BACKGROUND: Food waste is a global and growing problem that is gaining traction due to its environmental, ethical, social, and economic repercussions. Between 2022 and 2027, the worldwide papaya market is expected to have a huge increase, meaning a growth in organic waste, including peels and seeds. Thus, this study evaluated the potential use of peels and seeds of two mature papaya fruits as a source of bioactive compounds, converting these by-products into value-added products. Proximate analysis (AOAC methods), mineral content (ICP-MS), free sugars (HPLC-ELSD), fatty acid composition (GC-FID), vitamin E profile (HPLC-DAD-FLD), and antioxidant activity (DPPH and FRAP assays) were evaluated. RESULTS: Both by-products showed high total protein (20-27%), and dietary fiber (32-38%) contents. Papaya peels presented a high ash content (14-16%), indicating a potential application as a mineral source. 14 fatty acids were detected, with α-linolenic acid (30%) as the most abundant in the peels and oleic acid (74%) in the seeds. Both by-products showed high antioxidant activity. CONCLUSION: Papaya by-products display great potential for industrial recovery and application, such as formulation of new functional food ingredients.

Use of a Pteridine Moiety to Track DNA Uptake in Cells.

Fluorescence labeled oligonucleotides have a long history of being used to monitor nucleic acid transport and uptake. However, it is not known if the fluorescent moiety itself physically limits the number of pathways that can be used by the cell due to steric, hydrophobic, or other chemical characteristics. Here, we report a method for comparing the uptake kinetics of oligonucleotides labeled either with the fluorescent pteridine, 3-methyl-8-(2-deoxy-ß-D-ribofuranosyl) isoxanthopterin (3MI), or the common fluorophore 5-carboxyfluorescein (5-FAM). We use a multiphoton microscopic technique to monitor nucleic acid uptake LLC-PK1, a pig renal tubular cell line that is known to have multiple uptake pathways. We find that the two fluorophores enter the cells at different rates, suggesting that choice of fluorescent moiety influences the uptake pathway used by a cell. Finally, we reconstituted an LLC-PK1 membrane channel that is selective for nucleic acids in planar lipid bilayers, and tested the ability of the labeled nucleic acids to permeate the channel. We find that 3MI, and not 5-FAM labeled oligonucleotides can traverse the plasma membrane through the channel. These results have implications for future studies aimed at delivering pteridine moieties to cells and for tracking nucleic acid transport into tissues.

Further Evidence on Trace Element Imbalances in Haemodialysis Patients-Paired Analysis of Blood and Serum Samples.

Previous studies have shown that haemodialysis patients have an increased risk of trace element imbalances. Most studies have determined the concentration of trace elements in serum only, but most trace elements are not uniformly distributed between plasma and blood cells, which justifies separate analysis of the different compartments. In this study, we determined both the serum and whole blood concentration of a wide panel of trace elements (Li, B, Mn, Co, Ni, Cu, Zn, Se, Rb, Sr, Mo, Cd, Pb) in haemodialysis patients and compared them with those of a control group. Whole blood and serum samples were collected during routine laboratory testing of patients undergoing chronic haemodialysis. For comparison purposes, samples from individuals with normal renal function were also analysed. Statistically significant differences (p < 0.05) were found between the two groups for whole blood concentrations of all analysed elements except Zn (p = 0.347). For serum, the difference between groups was statistically significant for all elements (p < 0.05). This study confirms that patients on haemodialysis tend to present significant trace element imbalances. By determining the concentration of trace elements in both whole blood and serum, it was shown that chronic haemodialysis may affect intra- and extracellular blood compartments differently.

Otolith shape and elemental signatures provide insights into the connectivity of euryhaline Chelon labrosus inhabiting two close estuaries with different burdens of xenoestrogens in the Southern Bay of Biscay.

Intersex gonads have been observed in thicklip grey mullet Chelon labrosus, inhabiting estuaries with high burdens of xenoestrogens in the Southeast Bay of Biscay, but knowledge of population connectivity among estuaries is lacking for this euryhaline fish species. This study investigates the population structure of C. labrosus using otolith shape and elemental signatures of 60 adults (overall length âˆ¼ 38 cm) from two estuaries 21 nautic miles apart, one with a high incidence of intersex condition (Gernika), and the other one pristine (Plentzia). Otolith shape analyses were performed using elliptical Fourier descriptors, while elemental signatures of whole sagittae were obtained by inductively coupled plasma mass spectrophotometry. Univariate and multivariate statistics were applied to determine if otolith signatures show patterns of homogeneity between estuaries. The data indicated significant differences in the otolith shape and elemental composition between mullets of Gernika and Plentzia. Elemental differences were mainly driven by Sr, Li (both higher in Plentzia) and Ba (higher in Gernika). The 98% re-classification success rate obtained from stepwise linear discriminant function analysis suggests that Gernika and Plentzia individuals form separated population units. The limited connectivity between these two close estuaries would indicate a different life history of exposure to chemicals, which might explain the high prevalence of intersex condition in Gernika and its absence in Plenztia.

Serum Iodine and Bromine in Chronic Hemodialysis Patients-An Observational Study in a Cohort of Portuguese Patients.

Background: Patients on chronic hemodialysis therapy are at high risk of disturbances in trace element status due to both the underlying disease and the hemodialysis process itself. Data on iodine and bromine levels in these patients are scarce. Methods: Using an ICP-MS analytical procedure, serum iodine and bromine levels were determined in a cohort (n = 57) of end-stage renal disease patients on chronic hemodialysis. The results were compared with those of a control group (n = 59). Results: Hemodialysis patients presented serum iodine levels within the normal range, slightly lower than in controls, but without reaching a statistically significant difference (67.6 ± 17.1 µg/L vs. 72.2 ± 14.8 µg/L; p = 0.1252). In contrast, serum bromine levels were much lower in patients (1086 ± 244 µg/L vs. 4137 ± 770 µg/L; p < 0.0001), at values only about 26% of the values observed in controls. Conclusions: Hemodialysis patients had normal serum iodine levels, but highly decreased serum bromine levels. The clinical significance of this finding requires further investigation, but it may be associated with sleep disturbances and fatigue that affect hemodialysis patients.

Placental levels of essential and non-essential trace element in relation to neonatal weight in Northwestern Spain: application of generalized additive models.

Adequate gestational progression depends to a great extent on placental development, which can modify maternal and neonatal outcomes. Any environmental toxicant, including metals, with the capacity to affect the placenta can alter the development of the pregnancy and its outcome. The objective of this study was to correlate the placenta levels of 14 essential and non-essential elements with neonatal weight. We examined relationships between placental concentrations of arsenic, cadmium, cobalt, copper, mercury, lithium, manganese, molybdenum, nickel, lead, rubidium, selenium, strontium, and zinc from 79 low obstetric risk pregnant women in Ourense (Northwestern Spain, 42°20'12.1″N 7°51.844'O) with neonatal weight. We tested associations between placental metal concentrations and neonatal weight by conducting multivariable linear regressions using generalized linear models (GLM) and generalized additive models (GAM). While placental Co (p = 0.03) and Sr (p = 0.048) concentrations were associated with higher neonatal weight, concentrations of Li (p = 0.027), Mo (p = 0.049), and Se (p = 0.02) in the placenta were associated with lower newborn weight. Our findings suggest that the concentration of some metals in the placenta may affect fetal growth.

First approach to assess the effects of nanoplastics on the soil species Folsomia candida: A mixture design with bisphenol A and diphenhydramine.

The terrestrial environment is one of the main recipients of plastic waste. However, limited research has been performed on soil contamination by plastics and even less assessing the effects of nanoplastics (NPls). Behind the potential toxicity caused per se, NPls are recognized vectors of other environmental harmful contaminants. Therefore, the main aim of the present study is to understand whether the toxicity of an industrial chemical (bisphenol A - BPA) and a pharmaceutical (diphenhydramine - DPH) changes in the presence of polystyrene NPls to the terrestrial invertebrate Folsomia candida. Assessed endpoints encompassed organismal (reproduction, survival and behavior) and biochemical (neurotransmission and oxidative stress) levels. BPA or DPH, 28 d single exposures (1 to 2000 mg/kg), induce no effect on organisms' survival. In terms of reproduction, the calculated EC50 (concentration that causes 50% of the effect) and determined LOEC (lowest observed effect concentration) were higher than the environmental concentrations, showing that BPA or DPH single exposure may pose no threat to the terrestrial invertebrates. Survival and reproduction effects of BPA or DPH were independent on the presence of NPls. However, for avoidance behavior (48 h exposure), the effects of the tested mixtures (BPA + NPls and DPH + NPls) were dependent on the NPls concentration (at 0.015 mg/kg - interaction: no avoidance; at 600 mg/kg - no interaction: avoidance). Glutathione S-transferase activity increased after 28 d exposure to 100 mg/kg DPH + 0.015 mg/kg NPls (synergism). The increase of lipid peroxidation levels found after the exposure to 0.015 mg/kg NPls (a predicted environmental concentration) was not detected in the mixtures (antagonism). The results showed that the effects of the binary mixtures were dependent on the assessed endpoint and the tested concentrations. The findings of the present study show the ability of NPls to alter the effects of compounds with different natures and mechanisms of toxicity towards soil organisms, showing the importance of environmental risk assessment considering mixtures of contaminants.

Uncovering the effects of copper feed supplementation on the selection of copper-tolerant and antibiotic-resistant Enterococcus in poultry production for sustainable environmental practices.

The use of antibiotics in animal production is linked to the emergence and spread of antibiotic-resistant bacteria, a threat to animal, environmental and human health. Copper (Cu) is an essential element in poultry diets and an alternative to antibiotics, supplementing inorganic or organic trace mineral feeds (ITMF/OTMF). However, its contribution to select multidrug-resistant (MDR) and Cu tolerant Enterococcus, a bacteria with a human-animal-environment-food interface, remains uncertain. We evaluated whether feeding chickens with Cu-ITMF or Cu-OTMF contributes to the selection of Cu tolerant and MDR Enterococcus from rearing to slaughter. Animal faeces [2-3-days-old (n = 18); pre-slaughter (n = 16)] and their meat (n = 18), drinking-water (n = 14) and feed (n = 18) from seven intensive farms with ITMF and OTMF flocks (10.000-64.000 animals each; 2019-2020; Portugal) were sampled. Enterococcus were studied by cultural, molecular and whole-genome sequencing methods and Cu concentrations by ICP-MS. Enterococcus (n = 477; 60 % MDR) were identified in 80 % of the samples, with >50 % carrying isolates resistant to tetracycline, quinupristin-dalfopristin, erythromycin, streptomycin, ampicillin or ciprofloxacin. Enterococcus with Cu tolerance genes, especially tcrB ± cueO, were mainly found in faeces (85 %; E. faecium/E. lactis) of ITMF/OTMF flocks. Similar occurrence and load of tcrB ± cueO Enterococcus in the faeces was detected throughout the chickens' lifespan in the ITMF/OTMF flocks, decreasing in meat. Most of the polyclonal MDR Enterococcus population carrying tcrB ± cueO or only cueO (67 %) showed a wild-type phenotype (MICCuSO4 ≤ 12 mM) linked to absence of tcrYAZB or truncated variants, also detected in 85 % of Enterococcus public genomes from poultry. Finally, < 65 µg/g Cu was found in all faecal and meat samples. In conclusion, Cu present in ITMF/OTMF is not selecting Cu tolerant and MDR Enterococcus during chickens' lifespan. However, more studies are needed to assess the minimum concentration of Cu required for MDR bacterial selection and horizontal transfer of antibiotic resistance genes, which would support sustainable practices mitigating antibiotic resistance spread in animal production and the environment beyond.

The Improved Quality of Gluten-Free Bread Due to the Use of Flaxseed Oil Cake: A Comprehensive Study Evaluating Nutritional Value, Technological Properties, and Sensory Quality.

The development of gluten-free bakery products, the quality of which is comparable to the quality of regular wheat-based products, remains a technological challenge. In this study, flaxseed oil cake (FOC), a by-product of flaxseed oil extraction and a source of nutritional and functional compounds, was used as an ingredient in the experimental bread formulation as partial replacement of starches (5%, 15%, and 30%). The gluten-free breads (GFBs) were evaluated for technological parameters, nutritional value, and sensory quality. Compared with the control, all FOC-enhanced GFBs were significantly (p < 0.05) richer in proteins, fat, and dietary fibre, with an increase that was proportional to the concentration of FOC in the formulation. At low-to-moderate levels (5% and 15%), FOC improved the specific volume, texture characteristics (reduced crumb hardness, gumminess, and chewiness), and appearance of GFBs, which allowed us to ameliorate its sensory features, but at 30% of FOC, the quality of the crumb texture decreased significantly (p < 0.005). Among the experimental GFBs, FOC15% exhibited improved technological characteristics and was rated by the sensory panel as the best in terms of overall quality. The results of the conducted research highlighted the benefits of incorporating FOC into GFB as a promising approach to developing a palatable, high-quality bakery product that may be a healthier option for individuals on a gluten-free diet, offering nutritional benefits. Nevertheless, it should be emphasised that the amount of FOC in GFB requires careful regulation.