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.

A commercial blend of macroalgae and microalgae promotes digestibility, growth performance, and muscle nutritional value of European seabass (Dicentrarchus labrax L.) juveniles.

Algae can leverage aquaculture sustainability and improve the nutritional and functional value of fish for human consumption, but may pose challenges to carnivorous fish. This study aimed to evaluate the potential of a commercial blend of macroalgae (Ulva sp. and Gracilaria gracilis) and microalgae (Chlorella vulgaris and Nannochloropsis oceanica) in a plant-based diet up to 6% (dry matter basis) on digestibility, gut integrity, nutrient utilization, growth performance, and muscle nutritional value of European seabass juveniles. Fish (11.3 ± 2.70 g) were fed with isoproteic, isolipidic, and isoenergetic diets: (i) a commercial-type plant-based diet with moderate fishmeal (125 g kg-1 DM basis) and without algae blend (control diet; Algae0), (ii) the control diet with 2% algae blend (Algae2), (iii) the control diet with 4% algae blend (Algae4), and (iv) the control diet with 6% algae blend (Algae6) for 12 weeks. The digestibility of experimental diets was assessed in a parallel study after 20 days. Results showed that most nutrients and energy apparent digestibility coefficients were promoted by algae blend supplementation, with a concomitant increase in lipid and energy retention efficiencies. Growth performance was significantly promoted by the algae blend, the final body weight of fish fed Algae6 being 70% higher than that of fish fed Algae0 after 12 weeks, reflecting up to 20% higher feed intake of algae-fed fish and the enhanced anterior intestinal absorption area (up to 45%). Whole-body and muscle lipid contents were increased with dietary algae supplementation levels by up to 1.79 and 1.74 folds in Algae 6 compared to Algae0, respectively. Even though the proportion of polyunsaturated fatty acids was reduced, the content of EPA and DHA in the muscle of algae-fed fish increased by nearly 43% compared to Algae0. The skin and filet color of juvenile European seabass were significantly affected by the dietary inclusion of the algae blend, but changes were small in the case of muscle, meeting the preference of consumers. Overall results highlight the beneficial effects of the commercial algae blend (Algaessence®) supplementation in plant-based diets for European seabass juveniles, but feeding trials up to commercial-size fish are needed to fully assess its potential.

Effect of Dietary Inorganic and Chelated Trace Mineral Supplementation on the Growth Performance and Skeletal Deformities of European Seabass and Senegalese Sole Post-larvae.

Trace elements such as Cu, Fe, Mn and Zn are essential minerals in fish diets, especially important at early larval stages. The chemical speciation of these elements directly influences their uptake efficiency and metabolic utilization. In order to optimize the form of trace elements incorporated into larval feed, two experiments were conducted using two commercial fish species, European seabass (Dicentrarchus labrax) and Senegalese sole (Solea senegalensis), and two chemical forms (inorganic and glycinate chelates). Several fish performance parameters were measured, as well as bone status parameters to assess which form of mineral results in optimal fish biological performance. European seabass and Senegalese sole post-larvae were unresponsive (P > 0.05) to dietary treatments in terms of dry weight (DW), standard length (SL), relative growth rate (RGR) or feed conversion rates (FCR) when fed diets supplemented with chelated over inorganic trace minerals. This study suggests that replacing dietary inorganic mineral supplementation by their organic glycinate-chelated forms brings no beneficial effects on somatic growth and bone development in Senegalese sole and European seabass post-larvae fed high-quality commercial microdiets. Additionally, we show that mineral leaching from diets can be significant, but the use of chelated minerals can potentially mitigate this leaching phenomenon. Therefore, the selection of the dietary mineral form should take into account not only their economic value, but also their biological effect and environmental impact. Data generated in this trial provides new knowledge in trace mineral nutrition of early-stage marine fish.

Association between Elevated Iodine Intake and IQ among School Children in Portugal.

The goal of this work was to examine whether elevated iodine intake was associated with adverse effects on IQ among school-age children in Portugal. In a representative sample of children from the north of the country, IQ percentiles by age (assessed with Raven's Colored Progressive Matrices) were dichotomized to <50 ("below-average" IQs) and ≥50. Morning urine iodine concentrations, corrected for creatinine, were dichotomized to <250 µg/g and ≥250 µg/g, according to the European Commission/Scientific Committee on Food's tolerable upper level of daily iodine intake for young children. Data were examined with Chi-square tests, logistic regression, and GLM univariate analysis. The sample (N = 1965) was classified as generally iodine-adequate (median urinary iodine concentration = 129 µg/L; median iodine-to-creatinine ratio = 126 µg/g) according to the WHO's criteria. A greater proportion of children in the ≥250 µg/g group had below-average IQs, compared to children with less than 250 µg/g (p = 0.037), despite a sizable (though non-significant) proportion of children in the less-than-250 µg/g group also presenting below-average IQs, at the bottom of the iodine distribution (<50 µg/g). The proportion of below-average IQs increased with increasingly elevated iodine concentrations (p = 0.047). The association remained significant after the adjustment for confounders, with the elevated iodine group showing increased odds of having below-average IQs when compared with the non-elevated iodine group (OR 1.55; 95% CI 1.11−2.17; p = 0.011). Consistently, the former group presented a lower mean IQ than the latter (p = 0.006). High iodine intake was associated with lower IQs even in a population classified as iodine-adequate. These results bear on child cognition and on initiatives involving iodine supplementation.

Trace Elements in Medicinal Plants Traditionally Used in the Treatment of Diabetes-Do They Have a Role in the Claimed Therapeutic Effect?

Background: Medicinal plants are often used in the treatment of diabetes mellitus, although knowledge about their mode of action and the substances responsible for their antidiabetic potential is limited. It is well known that some trace elements play a role in glucose metabolism and insulin action. Thus, a particular trace elements profile could be associated with the antidiabetic properties observed for some medicinal plants. Methods: Infusions (n = 102) prepared from commercial herbal products (n = 34) containing medicinal plants indicated for the treatment of diabetes (n = 16 different plant species) and infusions (n = 60) prepared from commercial herbal products (n = 20) containing medicinal plants without such an indication (n = 7 different plant species) were analyzed by ICP-MS for their trace elements content. Results: In both groups, results varied significantly between different medicinal plants and also between different origins (brands) of the same medicinal plant. Significant differences (p < 0.05) between the two groups were found for nine elements, including four trace elements related to glucose metabolism (Mn, B, V, and Se), but with lower median contents in the group of medicinal plants for diabetes. Conclusions: Except for some particular species (e.g., Myrtilli folium) in which the trace element Mn may play a role in its antidiabetic effect, globally, a direct association between the claimed antidiabetic properties and a specific trace element profile of the studied medicinal plants was not evident.

Seaweeds rehydration and boiling: Impact on iodine, sodium, potassium, selenium, and total arsenic contents and health benefits for consumption.

Considering the importance of seaweeds for the development of sustainable and innovative food products, this study aimed to characterize the impact of hydrothermal processing on iodine, sodium, potassium, selenium, and arsenic concentrations of four seaweed species (S. latissima, L. digitata, U. pinnatifida, and C. crispus) and on the associated health risks-benefits for consumers. These elements revealed a common pattern for leachable fractions of iodine, total arsenic, and selenium: L. digitata ≥ S. latissima > C. crispus > U. pinnatifida after rehydration and boiling during different periods. The behavior for sodium was: S. latissima > L. digitata > C. crispus > U. pinnatifida, and for potassium: U. pinnatifida > L. digitata > S. latissima > C. crispus. Generally, the species that attained more significant losses were S. latissima and L. digitata. A health-relevant sodium/potassium ratio below 0.7 was found for all species except for U. pinnatifida. In some species, the risk-benefit analysis revealed that high iodine and arsenic levels might promote risks for consumption, even after 20 min boiling, but 5 g of processed U. pinnatifida could contribute to adequate iodine, sodium, potassium, and selenium intakes for all population groups. Standardized processing treatments of seaweeds can open new opportunities for the sector.

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.

Effect of Dietary Manganese and Zinc Levels on Growth and Bone Status of Senegalese Sole (Solea senegalensis) Post-Larvae.

Essential dietary trace elements, such as zinc (Zn) and manganese (Mn), critically influence a wide range of physiological, metabolic, and hormonal processes in fish larvae and post-larvae. Despite their importance for normal fish growth and skeletal development, trace mineral nutrition has not been extensively studied in the early stages of development of fish. Post-larvae of an emergent aquaculture species, Senegalese sole (Solea senegalensis), were the subject of this study in order to better understand the effects of diet supplementation of trace minerals upon fish larval development and performance. Sole post-larvae were fed a combination of organic Mn (45 and 90 mg kg-1 feed) and organic Zn (100 and 130 mg kg-1 feed) and survival, growth, mineral deposition rates, and vertebral bone status were assessed. Our results showed that although no significant effect was found on the growth performance of Senegalese sole post-larvae, Mn and Zn supplementation to a commercial microdiet for marine fish larvae at higher dietary levels (Mn at 90 mg kg-1 and Zn at 130 mg kg-1) improved larval survival, decreased the severity of vertebral malformations, and increased the deposition of Mn in bone.

Effect of Zinc Source and Exogenous Enzymes Supplementation on Zinc Status in Dogs Fed High Phytate Diets.

Zinc is an essential element, a cofactor of many enzymes, and performs catalytic, structural and regulatory functions. Once in the gastrointestinal tract, zinc can interact with food constituents. Phytic acid, the major phosphorus storage in plants, limits zinc availability from animal feeds due to the formation of insoluble complexes with phytates. This study tested the effect of supplemental zinc source (zinc sulfate and a chelate zinc proteinate) and the addition of exogenous enzymes from a solid-state fermentation product of Aspergillus niger to a high phytate diet. The study was designed according to three Latin Squares 4 × 4 with a 2 × 2 factorial arrangement of treatments, with four periods, four diets, and 12 young adult Beagles. Periods lasted 5 weeks each. Diets were supplemented with 75 mg/kg of zinc sulfate (IZ) or zinc proteinate (OZ), and without or with 200 mg/kg of exogenous enzymes (IZ+, OZ+). Results showed that zinc proteinate increased the bioavailability of phosphorus, yet the zinc biomarkers remained unaffected by the zinc source, with the exception of lymphocyte subsets that benefit from zinc proteinate. The use of exogenous enzymes did not affect zinc availability nor nutrient and energy digestibility.

Nutritional, chemical and antioxidant/pro-oxidant profiles of silverskin, a coffee roasting by-product.

Coffee silverskin (a coffee roasting by-product) contains high amounts of dietary fibre (49% insoluble and 7% soluble) and protein (19%). Potassium (∼5g/100g), magnesium (2g/100g) and calcium (0.6g/100g) are the major macrominerals. The vitamin E profile of silverskin comprises α-tocopherol, ß-tocopherol, É£-tocopherol, δ-tocopherol, ß-tocotrienol, É£-tocotrienol, and δ-tocotrienol. The fatty acid profile is mainly saturated (C16:0 and C22:0), but the total amount of fat is low (2.4%). Caffeine (1.25g/100g), chlorogenic acid (246mg/100g), and 5-hydroxymethylfurfural (5.68mg/100g) are also present in silverskin. Total phenolics and flavonoids are partially responsible for the in vitro antioxidant activity. Silverskin extracts protected erythrocytes from oxidative AAPH- and H2O2-induced hemolysis, but at high concentrations a pro-oxidant effect on erythrocyte morphology was observed.

Mineral Composition of Dry Dog Foods: Impact on Nutrition and Potential Toxicity.

Detailed mineral profile of a selection of commercially available complete dry dog foods was determined using ICP-MS (Se, Cu, Mn, Zn, and nonessential trace elements), flame photometry (Na and K) and atomic and molecular spectrophotometry (Ca, P, Mg, and Fe). The contribution of ingredients to the mineral composition was correlated to the food market segment. Results showed an oversupply of essential elements due to the energy density effect on feed intake. Additives contributed from 40.8 to 55.1% to the total trace elements contents. With the exception of Se, all trace elements were supplied above the nutritional requirements of adult dogs. Legal limits of Cu, Se, and Zn were surpassed. The content of nonessential trace elements included values in the range of nanograms to micrograms per kg, without surpassing safe upper limits. This work brings awareness to the need to find supplementation strategies that ensure nutritional adequacy and avoid waste.

Fibre fortification of wheat bread: impact on mineral composition and bioaccessibility.

In this work, wheat bread was fortified with fibre enriched extracts recovered from agroindustry by-products, namely, elderberry skin, pulp and seeds (EE); orange peel (OE); pomegranate peel and interior membranes (PE); and spent yeast (YE). The impact of this fortification on the total and bioaccessible mineral composition of wheat breads, estimated mineral daily intake, and the relationship between bioaccessibility and dietary fibre was evaluated. Fortification with OE, EE, and PE improved the content of essential minerals in bread when compared to control bread. The exception was bread fortified with YE, which presented a mineral content similar to control bread, but its mineral bioaccessibility was significantly higher than in all the other bread formulations. The opposite was observed for PE bread, which presented a significant reduction of bioaccessible minerals. We concluded that the origin of the fibre rich extract must be carefully selected, to avoid potential negative impact on mineral bioaccessibility.

Anthropogenic pressure in a Portuguese river: Endocrine-disrupting compounds, trace elements and nutrients.

Natural organic compounds such as phytoestrogens and phytosterols found in various plants, as well as mycotoxins produced by fungi, can be found in aquatic environments. The aim of this study was to investigate the occurrence of three different classes of natural estrogenic compounds, i.e., phytoestrogens, phytosterols and mycotoxins, in estuarine water samples from the Ave River estuary. For that, water samples were collected at five sampling points distributed along the estuary at low tide, during 1 year, processed by solid-phase extraction (SPE) and analyzed by gas chromatography coupled to mass spectrometry (GC-MS). To correlate the presence of phytoestrogens and phytosterols in the estuarine water, local flora was collected on riverside. Trace elements content and physicochemical parameters such as nutrients and dissolved oxygen were also determined seasonally at each sampling point, to give insights for the evaluation of water quality and anthropogenic pressure. Both phytoestrogens and phytosterols showed a seasonal variation, with the highest values observed in spring and summer and the lowest in winter. Daidzein (DAID) was found up to 404.0 ng L(-1) in spring and coumestrol (COUM) was found up to 165.0 ng L(-1) in summer. The mycotoxin deoxynivalenol (DON) was ubiquitously determined with values ranging from 59.5 to 642.4 ng L(-1). Nutrients and metals distribution and concentration varied among sampling stations and seasons. This study revealed for the first time the presence of mycotoxins, various classes of phytoestrogens and stigmasterol (STG) in estuarine water from the Ave River (Portugal), and the evaluation of the water quality confirmed that this estuary is still highly impacted by anthropogenic activities.

Anatomical regional differences in selenium levels in the human brain.

The role of selenium in human brain physiology, as well as in aging and neurodegenerative processes, remains unclear. Thus, the aim of this study was to establish the "normal" (reference) levels for selenium in the human brain, as well as anatomical regional differences and age-related changes. Using inductively coupled plasma-mass spectrometry after sample microwave-assisted acid digestion, selenium levels were measured in 14 different areas of the brain of adult individuals (n = 42; 71 ± 12, range 50-101 years old) without a known history of neurodegenerative, neurological, or psychiatric disorders. In the whole data set (n = 588; 42 individuals × 14 brain areas), selenium levels ranged from 552 to 1435 ng/g, with a mean ± SD content of 959 ± 178 ng/g (dry weight basis). Selenium distribution across the different brain areas was heterogeneous, with the highest levels in the putamen, parietal inferior lobule, and occipital cortex and the lowest expression in the medulla and cerebellum. Selenium levels were unchanged with aging. Compared with the age-matched control group, significantly increased levels of selenium were found in the globus pallidus, superior temporal gyrus, and frontal cortex of Parkinson's disease (n = 1) and Alzheimer's disease (n = 2) patients. This study provides new data on the anatomical regional differences in selenium levels in the human brain, which will aid future interpretation of studies examining brain tissue affected by neurodegenerative (and other) brain diseases.

Trace elements in human milk: correlation with blood levels, inter-element correlations and changes in concentration during the first month of lactation.

Using inductively coupled plasma mass spectrometry (ICP-MS) based analytical procedures, the concentration of several trace elements (Mn, As, Pb, Co, Ni, Cu, Zn and Se) was determined in human milk samples collected from a group of healthy lactating Portuguese women (n=44), both on the 2nd day postpartum (i.e., colostrum; n=34) and at 1 month postpartum (i.e., mature milk; n=19). Blood samples (n=44), collected on the 2nd day after parturition, were also analyzed for the same trace elements. No major correlations were observed between the levels of the analyzed trace elements in blood and colostrum samples. All the studied elements, except for Co, Pb and Ni, showed a significant trend for a decrease in concentration in milk during the first month of lactation. This trend was more pronounced for Zn and Se, whose levels decreased to approximately 23% and 44% of their initial mean concentration, respectively. With the exception of Co (r=0.607) and Zn (r=0.487), no significant correlations were observed when comparing the levels of each trace element between samples of colostrum and mature milk. Several inter-element correlations were found within each type of milk sample. The most significant were: (i) Se vs Cu (r=0.828) and Se vs Co (r=0.605) in colostrum samples and (ii) Ni vs Pb (r=0.756), Ni vs Mn (r=0.743) and Se vs Co (r=0.714) in mature milk samples. An inverse correlation between Zn and Se was also found in both types of milk sample; however, it only reached statistical significance for mature milk (r=-0.624).