Microfluidic Paper-Based Device Incorporated with Silica Nanoparticles for Iodide Quantification in Marine Source Dietary Supplements.

Iodine is an essential micronutrient for humans due to its fundamental role in the biosynthesis of thyroid hormones. As a key parameter to assess health conditions, iodine intake needs to be monitored to ascertain and prevent iodine deficiency. Iodine is available from various food sources (such as seaweed, fish, and seafood, among others) and dietary supplements (multivitamins or mineral supplements). In this work, a microfluidic paper-based analytical device (µPAD) to quantify iodide in seaweed and dietary supplements is described. The developed µPAD is a small microfluidic device that emerges as quite relevant in terms of its analytical capacity. The quantification of iodide is based on the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) by hydrogen peroxide in the presence of iodine, which acts as the catalyst to produce the blue form of TMB. Additionally, powder silica was used to intensify and uniformize the colour of the obtained product. Following optimization, the developed µPAD enabled iodide quantification within the range of 10-100 µM, with a detection limit of 3 µM, and was successfully applied to seaweeds and dietary supplements. The device represents a valuable tool for point-of-care analysis, can be used by untrained personnel at home, and is easily disposable, low-cost, and user-friendly.

Iodineminho Study: Iodine Supplementation and Prevalence of Iodine Deficiency in Pregnant Women.

CONTEXT: Iodine is necessary for the proper brain development. The prevalence of iodine deficiency in Portuguese pregnant women led the health authorities, in 2013, to recommend iodine supplementation for women in preconception, throughout pregnancy and during lactation. OBJECTIVE: To assess the impact of iodine supplementation initiated in the preconception or the first trimester of pregnancy on the prevalence of iodine deficiency and maternal thyroid status. METHODS: An observational prospective cohort study that follows thyroid function and iodine status of women recruited in preconception or in the first trimester of pregnancy. RESULTS: The median urinary iodine concentration (UIC) was significantly higher among women taking iodine supplements (no-supplement group UIC=63µg/L; supplement group UIC =100µg/L, p = 0.002) but still below the levels recommended by the World Health Organization. Only 15% of pregnant women had adequate iodine status and 17% showed UIC < 50 µg/l. There was no influence of whether iodine supplementation started in preconception or in the 1st trimester of gestation (UIC preconception group: 112µg/L vs UIC pregnancy group: 91µg/L, p = 0.569). In the 1st trimester of pregnancy, total thyroxine levels were lower and free triiodothyronine levels were higher in non-supplemented women. Thyroglobulin levels were lower in women who started iodine supplementation in preconception compared to non-supplemented women and women who started iodine supplementation during gestation. CONCLUSION: In the Minho region of Portugal, fertile women have insufficient iodine intake. Additional public health measures are needed since the current recommendations for iodine supplementation for pregnancy are unsatisfactory to achieve an adequate iodine status.

Iodine Availability through Iodized Salt in Portugal: 2010-2021 Sales Evolution and Distribution.

Salt iodization programs are considered the most cost-effective measures to ensure adequate iodine intake in iodine-deficient populations. Portuguese women of childbearing age and pregnant women were reported to be iodine-deficient, which led the health authorities, in 2013, to issue a recommendation for iodine supplementation during preconception, pregnancy and lactation. In the same year, iodized salt became mandatory in school canteens. Of note, no regulation or specific programs targeting the general population, or the impact of iodized salt availability in retailers, are known. The present study analyzed iodized salt supermarket sales from 2010 to 2021 from a major retailer, identifying the proportion of iodized salt in total salt sales and its distribution in mainland Portugal. Data on iodine content were collected through the nutritional label information. Of a total of 33 salt products identified, 3 were iodized (9%). From 2010 to 2021, the weighted sales of iodized salt presented a growing tendency, reaching the maximum of 10.9% of total sales (coarse plus fine salt) in 2021. Iodized salt reached a maximum of 11.6% of total coarse salt in 2021, a maximum of 2.4% of the total fine salt in 2018. The overall sales of iodized salt and their contribution to iodine intake are extremely low, prompting additional studies to understand the consumer's choice and awareness of the benefits of iodized salt.

Potential of phytoremediation for the removal of petroleum hydrocarbons in contaminated salt marsh sediments.

Degradation of petroleum hydrocarbons in colonized and un-colonized sediments by salt marsh plants Juncus maritimus and Phragmites australis collected in a temperate estuary was investigated during a 5-month greenhouse experiment. The efficiency of two bioremediation treatments namely biostimulation (BS) by the addition of nutrients, and bioaugmentation (BA) by addition of indigenous microorganisms was tested in comparison with hydrocarbon natural attenuation in un-colonized and with rhizoremediation in colonized sediments. Hydrocarbon degrading microorganisms and root biomass were assessed as well as hydrocarbon degradation levels. During the study, hydrocarbon degradation in un-colonized sediments was negligible regardless of treatments. Rhizoremediation proved to be an effective strategy for hydrocarbon removal, yielding high rates in most experiments. However, BS treatments showed a negative effect on the J. maritimus potential for hydrocarbon degradation by decreasing the root system development that lead to lower degradation rates. Although both plants and their associated microorganisms presented a potential for rhizoremediation of petroleum hydrocarbons in contaminated salt marsh sediments, results highlighted that nutrient requirements may be distinct among plant species, which should be accounted for when designing cleanup strategies.

Bioremediation potential of microorganisms from a sandy beach affected by a major oil spill.

The aim of this work was to evaluate the bioremediation potential of microorganisms from intertidal sediments of a sandy beach affected by a major oil spill 7 years before and subject to chronic petroleum contamination since then. For that, the response of microorganisms to a new oil contamination was assessed in terms of community structure, abundance, and capacity to degrade hydrocarbons. Experiments were carried out under laboratory-controlled conditions by mixing sediment with crude oil with three different nitrogen supplementations in 50 ml serum bottles under constant shake for 15 days. Autochthonous microorganisms were able to respond to the new oil contamination by increasing their abundance (quantified by DAPI) and changing the community structure (evaluated by DGGE). This response was particularly clear for some specific bacterial groups such as Pseudomonas, Actinomycetales, and Betaproteobacteria. These communities presented an important potential for hydrocarbon degradation (up to 85 % for TPHs and 70 % for total PAHs), being the biodegradation stimulated by addition of an appropriate amount of nitrogen.

Influence of different salt marsh plants on hydrocarbon degrading microorganisms abundance throughout a phenological cycle.

The influence of Juncus maritimus, Phragmites australis, and Triglochin striata on hydrocarbon degrading microorganisms (HD) in Lima River estuary (NW Portugal) was investigated through a year-long plant life cycle. Sediments un-colonized and colonized (rhizosediments) by those salt marsh plants were sampled for HD, total cell counts (TCC), and total petroleum hydrocarbons (TPHs) assessment. Generally, TCC seemed to be markedly thriving by the presence of roots, but without significant (p > 0.05) differences among rhizosediments. Nevertheless, plants seemed to have a distinct influence on HD abundance, particularly during the flowering season, with higher HD abundance in the rhizosediments of the fibrous roots plants (J. maritimus < P. australis < T. striata). Our data suggest that different plants have distinct influence on the dynamics of HD populations within its own rhizosphere, particularly during the flowering season, suggesting a period of higher rhizoremediation activity. Additionally, during the vegetative period, plants with fibrous and dense root system tend to retain hydrocarbons around their belowground tissues more efficiently than plants with adventitious root system. Overall results indicate that fibrous root plants have a higher potential to promote hydrocarbons degradation, and that seasonality should be taken into account when designing long-term rhizoremediation strategies in estuarine areas.

Bacterial community response to petroleum contamination and nutrient addition in sediments from a temperate salt marsh.

Microbial communities play an important role in the biodegradation of organic pollutants in sediments, including hydrocarbons. The aim of this study was to evaluate the response of temperate salt marsh microbial communities to petroleum contamination, in terms of community structure, abundance and capacity to degrade hydrocarbons. Sediments un-colonized and colonized (rhizosediments) by Juncus maritimus, Phragmites australis and Triglochin striata were collected in a temperate estuary (Lima, NW Portugal), spiked with petroleum under variable nutritional conditions, and incubated for 15 days. Results showed that plant speciation emerged as the major factor for shaping the rhizosphere community structure, overriding the petroleum influence. Moreover, when exposed to petroleum contamination, the distinct salt marsh microbial communities responded similarly with (i) increased abundance, (ii) changes in structure, and (iii) decreased diversity. Communities, particularly those associated to J. maritimus and P. australis roots displayed a potential to degrade petroleum hydrocarbons, with degradation percentages between 15% and 41%, depending on sediment type and nutritional conditions. In conclusion, distinct salt marsh microbial communities responded similarly to petroleum contamination, but presented different pace, nutritional requirements, and potential for its biodegradation, which should be taken into account when developing bioremediation strategies.

Potential of the microbial community present in an unimpacted beach sediment to remediate petroleum hydrocarbons.

The potential of the microbial communities present in the intertidal zone of an unimpacted beach (a beach that did not suffer any significant oil spill) to degrade hydrocarbons was investigated. For that, laboratory-based microcosms (50-ml flasks) were set up with sandy beach sediment spiked with crude oil and incubated with local seawater for 15 days in the dark. Three bioremediation treatments were tested (biostimulation (BS), autochthonous bioaugmentation (AB), and combined treatment of biostimulation + bioaugmentation (BS + AB)) and the results were compared with natural attenuation (NA). Visual inspection showed clearly an oil solubility increase (confirmed by a higher hydrocarbons concentration in supernatant solutions) for all tested treatments when compared to NA. Significant degradation of the oil, shown by different profiles of petroleum hydrocarbons, was also observed for the different treatments particularly for BS + AB. Therefore, the microbial community of this unimpacted beach sediment could respond to an oil spill, degrading hydrocarbons. But to increase the natural attenuation pace, obtained results indicated that BS + AB is an appropriate approach for the bioremediation of beaches recently impacted by an oil spill. The autochthonous microbial cultures can be obtained "before" or "after" the contamination of the target site, being inoculated into the site right after it contamination.

Biodegradation of petroleum hydrocarbons in estuarine sediments: metal influence.

In this work, the potential effect of metals, such as Cd, Cu and Pb, on the biodegradation of petroleum hydrocarbons in estuarine sediments was investigated under laboratory conditions. Sandy and muddy non-vegetated sediments were collected in the Lima River estuary (NW Portugal) and spiked with crude oil and each of the metals. Spiked sediments were left in the dark under constant shaking for 15 days, after which crude oil biodegradation was evaluated. To estimate microbial abundance, total cell counts were obtained by DAPI staining and microbial community structure was characterized by ARISA. Culturable hydrocarbon degraders were determined using a modified most probable number protocol. Total petroleum hydrocarbons concentrations were analysed by Fourier Transform Infrared Spectroscopy after their extraction by sonication, and metal contents were determined by atomic absorption spectrometry. The results obtained showed that microbial communities had the potential to degrade petroleum hydrocarbons, with a maximum of 32 % degradation obtained for sandy sediments. Both crude oil and metals changed the microbial community structure, being the higher effect observed for Cu. Also, among the studied metals, only Cu displayed measurable deleterious effect on the hydrocarbons degradation process, as shown by a decrease in the hydrocarbon degrading microorganisms abundance and in the hydrocarbon degradation rates. Both degradation potential and metal influence varied with sediment characteristics probably due to differences in contaminant bioavailability, a feature that should be taken into account in developing bioremediation strategies for co-contaminated estuarine sites.

Use of solid phase extraction for the sequential injection determination of alkaline phosphatase activity in dynamic water systems.

In this work, a solid phase extraction sequential injection methodology for the determination of alkaline phosphatase activity in dynamic water systems was developed. The determination of the enzymatic activity was based on the spectrophotometric detection of a coloured product, p-nitrophenol, at 405 nm. The p-nitrophenol is the product of the catalytic decomposition of p-nitrophenyl phosphate, a non-coloured substrate. Considering the low levels expected in natural waters and exploiting the fact of alkaline phosphatase being a metalloprotein, the enzyme was pre-concentrated in-line using a NTA Superflow resin charged with Zn(2+) ions. The developed sequential injection method enabled a quantification range of 0.044-0.441 unit mL(-1) of enzyme activity with a detection limit of 0.0082 unit mL(-1) enzyme activity (1.9 µmol L(-1) of pNP) and a determination rate of 17 h(-1). Recovery tests confirmed the accuracy of the developed sequential injection method and it was effectively applied to different natural waters and to plant root extracts.

Hydrocarbon degradation potential of salt marsh plant-microorganisms associations.

Estuaries are often considered sinks for contaminants and the cleanup of salt marshes, sensitive ecosystems with a major ecological role, should be carried out by means of least intrusive approaches, such as bioremediation. This study was designed to evaluate the influence of plant-microorganisms associations on petroleum hydrocarbons fate in salt marshes of a temperate estuary (Lima River, NW Portugal). Sediments un-colonized and colonized (rhizosediments) by different plants (Juncus maritimus, Phragmites australis, Triglochin striata and Spartina patens) were sampled in four sites of the lower and middle estuary for hydrocarbon degrading microorganisms (HD), total cell counts (TCC) and total petroleum hydrocarbons (TPHs) assessment. In general, TPHs, HD and TCC were significantly higher (P < 0.05) in rhizosediments than in un-colonized sediments. Also recorded were differences on the abundance of hydrocarbon degraders among the rhizosediment of the different plants collected at the same site (J. maritimus < P. australis < T. striata), with statistically significant differences (P < 0.05) between J. maritimus and T. striata. Moreover, strong positive correlations-0.81 and 0.84 (P < 0.05), between biotic (HD) and abiotic (organic matter content) parameters and TPHs concentrations were also found. Our data clearly suggest that salt marsh plants can influence the microbial community, by fostering the development of hydrocarbon-degrading microbial populations in its rhizosphere, an effect observed for all plants. This effect, combined with the plant capability to retain hydrocarbons around the roots, points out that salt marsh plant-microorganisms associations may actively contribute to hydrocarbon removal and degradation in estuarine environments.

Comparison of the response of three microalgae species exposed to elutriates of estuarine sediments based on growth and chemical speciation.

The elutriate sediment toxicity test (ESTT) provides a measure of the amount of a substance that is exchanged between the sediment and the aqueous phase during resuspension processes such as floods or dredging operations. This study used ESTT with two complementary aims: a comparison of the elutriates of two estuarine sediments (anaerobic muddy [A] and aerobic sandy [B]) in terms of toxicity and a comparison of the response of three different microalgae (Emiliania huxleyi (coccolithophore), Dunaliella minuta (green alga), and Phaeodactylum tricornutum (diatom)) to each elutriate in terms of growth, heavy metals uptake, and organic ligands release or uptake in order to find eventual differences of sensitivity. The interpretation of the results was based on chemical speciation in the culture media. Both elutriates, particularly A, were much richer than seawater (control medium) in some heavy metals and organic ligands able to bind strongly heavy metals. Elutriate A slightly inhibited P. tricornutum growth but stimulated growth of E. huxleyi and D. minuta. Elutriate B stimulated the growth of the three algal species. Therefore, the diatom behaved differently from both the coccolithophore and the green alga. Strong complexation of trace metals by organic ligands could be the cause of absence of the metallic toxicity of the elutriates. Growth inhibition of P. tricornutum in elutriate A could be caused by ammonia-N and/or organic compounds. The concentration of the organic ligands decreased markedly (about 75%) in both elutriates after 10 d of incubation in contrast to the control culture, where their concentration increased about 50% because of exudation. This phenomenon was interpreted to result from ligand uptake by the algae, free or as metal complexes. This work demonstrated that beside the evaluation of toxicity of free heavy metals to alga species, the organic ligands must not be ignored. Depending on the amount of ligand present, the toxicity can be reduced (sequestration) or enhanced (better availability through uptake of metal-ligand complexes). Since the applied ESTT is a standard procedure (U.S. Environmental Protection Agency) for the evaluation of dredged material proposed for ocean disposal, it is necessary to discuss results obtained during toxicity tests with such elutriates in detail.