Earthworms and Grape Marc: Simultaneous Production of a High-Quality Biofertilizer and Bioactive-Rich Seeds

Winemaking produces annually millions of tons of grape marc as a byproduct, which is a revaluable resource having many potential uses, including a nutrient‐rich organic soil amendment. However, its application as untreated raw material can damage crops owing to the release of phytotoxic polyphenols. This agronomic problems can be minimized by vermicomposting, as earthworms can partly digest polyphenols. This chapter reports the results obtained in the processing of grape marc derived from white wine through vermicomposting on an industrial scale to yield both a high quality organic, polyphenol‐free fertilizer and grape seeds as a source of bioactive compounds. Vermicomposting reduced substantially the residue biomass. In a very short‐term, the process yielded a nutrient‐rich, microbiologically active and stabilized peat‐like material that can be easily separated from the seeds by sieving. The isolation of the seeds eliminates the polyphenol‐associated phytotoxicity from the vermicompost and left those seeds prepared to be easily processed to get different bioactive compounds, mainly rich‐polyphenols extracts but also rich‐fatty acids seed oil. The procedure described is effective, simple, environmental‐friendly and economical, and can easily be scaled up for industrial application yielding a variety of added‐value products from the initial grape marc

Itinerarios en la sala de matemáticas del Museo de la Ciencia. Actividades para Primaria

Utilizando como pretexto la sala de matemáticas del Museo de la Ciencia de Valladolid se analiza la importancia de las actividades complementarias para las matemáticas. Se estudian las competencias y se investiga como la manipulación, el juego y la motivación influye en el proceso de enseñanza-aprendizaje. La última parte del trabajo gira en torno a la realización de dos itinerarios dentro del contexto de los módulos propuestos por el museo. El primero se centra en la presentación del contenido y en el descubrimiento de nuevos aspectos de las matemáticas por parte del alumnado. El segundo presenta actividades para que sean los propios profesores los que organicen su visita según los contenidos académicos que se quieren trabajar.Grado en Educación Primari

Species-specific effects of epigeic earthworms on microbial community structure during first stages of decomposition of organic matter

Background: Epigeic earthworms are key organisms in organic matter decomposition because of the interactions they establish with microorganisms. The earthworm species and the quality and/or substrate availability are expected to be major factors influencing the outcome of these interactions. Here we tested whether and to what extent the epigeic earthworms Eisenia andrei, Eisenia fetida and Perionyx excavatus, widely used in vermicomposting, are capable of altering the microbiological properties of fresh organic matter in the short-term. We also questioned if the earthworm-induced modifications to the microbial communities are dependent on the type of substrate ingested. Methodology/Principal Findings: To address these questions we determined the microbial community structure (phospholipid fatty acid profiles) and microbial activity (basal respiration and microbial growth rates) of three types of animal manure (cow, horse and rabbit) that differed in microbial composition, after being processed by each species of earthworm for one month. No differences were found between earthworm-worked samples with regards to microbial community structure, irrespective of type of manure, which suggests the existence of a bottleneck effect of worm digestion on microbial populations of the original material consumed. Moreover, in mesocosms containing cow manure the presence of E. andrei resulted not only in a decrease in bacterial and fungal biomass, but also in a reduced bacterial growth rate and total microbial activity, while no such reduction was found with E. fetida and P. excavatus. Conclusions/Significance: Our results point to the species of earthworm with its associated gut microbiota as a strong determinant of the process shaping the structure of microbial communities in the short-term. This must nonetheless be weighed against the fact that further knowledge is necessary to evaluate whether the changes in the composition of microbiota in response to the earthworm species is accompanied by a change in the microbial community diversity and/or function.Ministerio de Ciencia e Innovación | Ref. CTM2009-0847

Profiling the Fatty Acids Content of Ornamental Camellia Seeds Cultivated in Galicia by an Optimized Matrix Solid-Phase Dispersion Extraction

Camellia (genus of flowering plants of fam. Theaceae) is one of the main crops in Asia, where tea and oil from leaves and seeds have been utilized for thousands of years. This plant is excellently adapted to the climate and soil of Galicia (northwestern Spain) and northern Portugal where it is grown not only as an ornamental plant, but to be evaluated as a source of bioactive compounds. In this work, the main fatty acids were extracted from Camellia seeds of four varieties of Camellia: sasanqua, reticulata, japonica and sinensis, by means of matrix-solid phase dispersion (MSPD), and analyzed by gas chromatography (GC) with MS detection of the corresponding methyl esters. MSPD constitutes an efficient and greener alternative to conventional extraction techniques, moreover if it is combined with the use of green solvents such as limonene. The optimization of the MSPD extraction procedure has been conducted using a multivariate approach based on strategies of experimental design, which enabled the simultaneous evaluation of the factors influencing the extraction efficiency as well as interactions between factors. The optimized method was applied to characterize the fatty acids profiles of four Camellia varieties seeds, allowing us to compare their fatty acid compositionThis research was supported by European Regional Development Fund 2007–2013 (FEDER), and project GPC2014/035 (Consolidated Research Groups Program of Galician Government Xunta de Galicia)S

High-Resolution Mass Spectrometry for the Comprehensive Characterization of Plant-Pigment-Based Tattoos and Dyes Formulations

There has been little research on emerging natural-pigment-based temporary tattoos and their constituents so far. This study focused on the characterization of natural henna and jagua tattoos and dyes in commercial samples. Sample manipulation and treatment were minimal to analyze natural pigments as they are marketed. The characterization of active compounds was performed by ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS). A novel untargeted characterization was also applied for these types of samples, searching for compounds ranging from the original ingredients to other additives of interest they may contain. The fact that these products may also be a source of unlabelled allergens was considered. This work showed that not all of the 34 samples analyzed contained their characteristic active ingredients, and in addition, 70 substances of a different nature were found. Therefore, a rapid and high-throughput methodology has been developed for the identification of the components of these dyes, providing a necessary approach for quality control or identification of suspicious substancesThis research was supported by projects EQC2018-005011-P (Ministry of Science, Innovation and Universities, Spain) and ED431 2020/06 (Xunta de Galicia). The authors belong to the CRETUS Strategic Partnership (ED431E 2018/01). All these programs are co-funded by FEDER (UE). L.R. acknowledges Xunta de Galicia for her predoctoral contract (ED481A-2018/227)S

Changes in chemical and microbiological properties of rabbit manure in a continuous-feeding vermicomposting system

In the present study the potential of the earthworm Eisenia fetida to process large amounts of waste was evaluated through continuous feeding reactors in which new layers of rabbit manure were added sequentially to form an age gradient inside the reactors. An optimal moisture level, ranging from 66% to 76%, was maintained throughout the process using an automatic watering system. The pH was close to 8.3, but decreased to 7.6 after 200 d of vermicomposting. No changes in electrical conductivity through the prole of layers were detected. Based on comparisons of phospholipid fatty acid (PLFA) proles and microbial activity measurements (basal respiration), a decrease in the levels of bacteria and fungi in layers corresponding to vermicomposting times of more than 200 d occurred. This points to a higher degree of stabilisation in the nal product, which is of utmost importance for its safe use as an organic amendment.Ministerio de Ciencia e Innovación | Ref. CTM2009-0847

Monitoring of Natural Pigments in Henna and Jagua Tattoos for Fake Detection

Temporary tattoos are a popular alternative to permanent ones. Some of them use natural pigments such as lawsone in the famous henna tattoos. Recently, jagua tattoos, whose main ingredients are genipin and geniposide, have emerged as an interesting option. This study was conducted to identify the presence and concentration of henna and jagua active ingredients (lawsone; genipin and geniposide, respectively) in commercial tattoo samples. Since natural pigments are often mixed with additives such as p-phenylenediamine (PPD) in the case of henna, PPD has been included in the study. Green and simple extraction methods based on vortex or ultrasound-assisted techniques have been tested. To determine the compounds of interest liquid chromatography (LC) with diode-array detection (DAD) has been applied; and PPD absence was confirmed by LC-QTOF (quadrupole-time of flight tandem mass spectrometry). This work demonstrated that only one out of 14 henna samples analyzed contained lawsone. For jaguas, genipin was found in all samples, while geniposide only in two. Therefore, quality control analysis on these semi-permanent tattoos is considered necessary to detect these ingredients in commercial mixtures, as well as to uncover possible fraud in products sold as natural hennaThis research was supported by project EQC2018-005011-P (Ministry of Science, Innovation and Universities, Spain. The authors belong to the CRETUS Strategic Partnership (ED431E2018/01). All these programmes are co-funded by FEDER (UE). L.R. acknowledges Xunta de Galicia for her predoctoral contractS

Recycling and valorization of distilled grape marc through vermicomposting: a pilot-scale study

The present study sought to evaluate the effectiveness of vermicomposting for the valorization of distilled grape marc, one of the main solid by-products of the winery sector during a 56-day pilot-scale trial. The increase in the density and biomass of earthworms ( Eisenia andrei ) during the earlier stages of the process reflected the suitability of the distilled marc ( Vitis vinifera L. cv. Mencía) as feedstock in order to sustain large earthworm populations on a pilot-scale level. Supporting this, from 14 days onwards the pH of Mencía distilled marc fell within weak-alkaline levels and the electrical conductivity was between 0.21 and 0.11 mS cm −2 providing optimum conditions for earthworm growth. A rapid decrease in microbial activity as well as in the content of total polyphenols, both indicative of stabilized materials was also recorded after 14 days of vermicomposting. Moreover, the content of macro- and micronutrients in the end product matched with those considered to have the quality criteria of a good vermicompost with respect to plant health and safe agricultural use. Altogether, it underlines the feasibility of vermicomposting as an environment-friendly approach for the biological stabilization of distilled grape marc fulfilling both environmental protection and fertilizer production.Agencia Estatal de Investigación | Ref. PID2021-124265OB-I00Xunta de Galicia | Ref. ED431C 2022/07Agencia Estatal de Investigación | Ref. RYC-2016-21231Universidade de Vigo/CISU

Epigeic earthworms exert a bottleneck effect on microbial communities through gut associated processes

Background: Earthworms play a critical role in organic matter decomposition because of the interactions they establish with microorganisms. The ingestion, digestion, assimilation of organic material in the gut and then casting is the first step in earthworm-microorganism interactions. The current knowledge of these direct effects is still limited for epigeic earthworm species, mainly those living in man-made environments. Here we tested whether and to what extent the earthworm Eisenia andrei is capable of altering the microbiological properties of fresh organic matter through gut associated processes; and if these direct effects are related to the earthworm diet. Methodology: To address these questions we determined the microbial community structure (phospholipid fatty acid profiles) and microbial activity (fluorescein diacetate hydrolysis) in the earthworm casts derived from three types of animal manure (cow, horse and pig manure), which differed in microbial composition. Principal Findings: The passage of the organic material through the gut of E. andrei reduced the total microbial biomass irrespective of the type of manure, and resulted in a decrease in bacterial biomass in all the manures; whilst leaving the fungi unaffected in the egested materials. However, unlike the microbial biomass, no such reduction was detected in the total microbial activity of cast samples derived from the pig manure. Moreover, no differences were found between cast samples derived from the different types of manure with regards to microbial community structure, which provides strong evidence for a bottleneck effect of worm digestion on microbial populations of the original material consumed. Conclusions/Significance: Our data reveal that earthworm gut is a major shaper of microbial communities, thereby favouring the existence of a reduced but more active microbial population in the egested materials, which is of great importance to understand how biotic interactions within the decomposer food web influence on nutrient cycling.Ministerio de Ciencia e Innovación | Ref. CTM2009-0847

Tracking Bacterial Spoilage in Cosmetics by a New Bioanalytical Approach: API-SPME-GC-MS to Monitor MVOCs

The main goal of this work was the use of the powerful solid-phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS) technique to unequivocally identify microbial volatile organic compounds (MVOCs) derived from the enzymatic activity produced during metabolic processes using analytical profile index (API) biochemical tests. Three bacteria were selected for this study: Escherichia coli, Proteus mirabilis, and Pseudomonas aeruginosa. They were inoculated and incubated to both API components and real cosmetics, as well as to a mixture of them. Specific MVOCs were successfully identified as biomarkers for each one of the studied microorganisms: Indole and 2-nitrophenol as Escherichia coli markers, 2-undecanone and phenylethyl alcohol as Proteus mirabilis-specific markers, and 1-undecene and 2′-aminoacetophenone as Pseudomonas aeruginosa ones. In addition, a high number of MVOCs were identified as general markers of bacterial presence. The results revealed that the MVOCs’ formation is highly subtract dependent. Therefore, the ultimate and most challenging objective is to establish a relationship between the identified MVOCs and the original compound present in the substrate. This work establishes the design and development of this original approach, and its practical application to the control of microbial contamination in real cosmetic samplesThis research was funded by project QFashion (IN852A 2016/157, CONECTA-PEME, Xunta de Galicia). The authors belong to the CRETUS Strategic Partnership (ED431E 2018/01). All these programmes are co-funded by FEDER (UE)S