In vivo toxicity assessment of eugenol and vanillin-functionalised silica particles using Caenorhabditis elegans.

The toxicological properties of different silica particles functionalised with essential oil components (EOCs) were herein assessed using the in vivo model C. elegans. In particular, the effects of the acute and long-term exposure to three silica particle types (SAS, MCM-41 micro, MCM-41 nano), either bare or functionalised with eugenol or vanillin, were evaluated on different biological parameters of nematodes. Acute exposure to the different particles did not reduce nematodes survival, brood growth or locomotion, but reproduction was impaired by all the materials, except for vanillin-functionalised MCM-41 nano. Moreover, long-term exposure to particles led to strongly inhibited nematodes growth and reproduction. The eugenol-functionalised particles exhibited higher functionalisation yields and had the strongest effects during acute and long-term exposures. Overall, the vanillin-functionalised particles displayed milder acute toxic effects on reproduction than pristine materials, but severer toxicological responses for the 96-hour exposure assays. Our findings suggest that the EOC type anchored to silica surfaces and functionalisation yield are crucial for determining the toxicological effects of particles on C. elegans. The results obtained with this alternative in vivo model can help to anticipate potential toxic responses to these new materials for human health and the environment.

Predicting Gilthead Sea Bream (Sparus aurata) Freshness by a Novel Combined Technique of 3D Imaging and SW-NIR Spectral Analysis.

A technique that combines the spatial resolution of a 3D structured-light (SL) imaging system with the spectral analysis of a hyperspectral short-wave near infrared system was developed for freshness predictions of gilthead sea bream on the first storage days (Days 0-6). This novel approach allows the hyperspectral analysis of very specific fish areas, which provides more information for freshness estimations. The SL system obtains a 3D reconstruction of fish, and an automatic method locates gilthead's pupils and irises. Once these regions are positioned, the hyperspectral camera acquires spectral information and a multivariate statistical study is done. The best region is the pupil with an R² of 0.92 and an RMSE of 0.651 for predictions. We conclude that the combination of 3D technology with the hyperspectral analysis offers plenty of potential and is a very promising technique to non destructively predict gilthead freshness.

Characterisation of chemical damage on tissue structures by multispectral imaging and machine learning procedures: Alkaline hypochlorite effect in C. elegans.

Multispectral imaging represents a powerful technique to maximise data collection and analysis for biological materials. It improves the exploitation and understanding of in vivo/vitro experiments. This work focused on testing the capability of multispectral imaging to characterise the tissue damage produced by alkaline hypochlorite on the body and eggs of the biological model C. elegans. To that end, three synchronisation processes with different final bleach and sodium hydroxide concentrations were performed. The impact of treatments was characterised by measuring egg viability and morphology, besides capturing multispectral images of both nematode bodies and eggs. Multispectral images consisted of seven slices captured from different wavelengths within the visible/infrared spectrum by different light-pass filters. The results showed dependence between increased alkaline hypochlorite concentration and loss of egg viability/morphology. This relation was also observed for the imaging data, which showed alterations to tissue transmittance for all the tested wavelengths for both bodies and eggs. Localised alterations related to alkaline hypochlorite diffusion through anatomical nematode orifices were recognised. Applying multivariate methods to imaging data successfully characterised tissue alterations, from which treatment type was predicted for both nematodes and eggs. Moreover, the alterations recorded by imaging data were also used to predict egg viability regardless of treatment type (0.94). The high correlation between the imaging data from nematodes and eggs with egg viability evidenced multispectral imaging's ability to characterise tissue damage and its possible practical application to study alterations to the tissues of this biological model.

Studying process variables to obtain undisturbed shaped soft meat for people with poor oral health.

This study evaluated injection (I) and vacuum impregnation (VI) as the best methods to apply papain, and other important processing conditions (batch, aging and cooking conditions), to obtain soft meat (suitable for people with poor oral health) without disturbing its original shape. Two aging times were evaluated and four cooking conditions by immersion in soup. Meat samples were injected or vacuum-impregned (0.85 kPa) with a papain solution (5% w/v). After cooking, they were analyzed by the compression test, and by image and sensory analyses. The results indicated that by using both methods to apply the enzyme, the obtained meat was suitable for people with poor oral health, even if VI was the better method because it minimized the factors batch, aging and cooking condition. Therefore, the best meat processing method to obtain panelists' highest softness values and the best appreciation was employing aged meat pretreated by VI and cooked at 65 °C for 10 min.

Effects of essential oil components exposure on biological parameters of Caenorhabditis elegans.

The extensive use of essential oil components in an increasing number of applications can substantially enhance exposure to these compounds, which leads to potential health and environmental hazards. This work aimed to evaluate the toxicity of four widely used essential oil components (carvacrol, eugenol, thymol, vanillin) using the in vivo model Caenorhabditis elegans. For this purpose, the LC50 value of acute exposure to these components was first established; then the effect of sublethal concentrations on nematodes' locomotion behaviour, reproduction, heat and oxidative stress resistance and chemotaxis was evaluated. The results showed that all the components had a concentration-dependent effect on nematode survival at moderate to high concentrations. Carvacrol and thymol were the two most toxic compounds, while vanillin had the mildest toxicological effect. Reproduction resulted in a more sensitive endpoint than lethality to evaluate toxicity. Only pre-exposure to carvacrol and eugenol at the highest tested sublethal concentrations conferred worms oxidative stress resistance. However, at these and lower concentrations, both components induced reproductive toxicity. Our results evidence that these compounds can be toxic at lower doses than those required for their biological action. These findings highlight the need for a specific toxicological assessment of every EOC application.

Evaluation of the influence of food intake on the incorporation and excretion kinetics of mesoporous silica particles in C.elegans.

The effect of the presence of food on the incorporation and excretion of silica particles was studied in this work using the biological model Caenorhabditis elegans and image analysis techniques. The experiment was based on two 24-hour phases: exposure and depuration. During exposure, nematodes were maintained for 24 h in liquid medium with silica particles, but some with and others without food. During depuration, nematodes were transferred to medium without particles. Nematodes were analysed by an image analysis in both phases to quantify the properties of particle distributions in nematodes' bodies with time. No differences were found in the proportion of nematodes carrying particles in the exposure phase when food was present. However in the depuration phase, lack of food generated a high proportion of particle carriers. Particle distribution properties were also similar in the exposure phase. Nevertheless, lack of food produced particle accumulation due to decelerated excretion because digestive tube relaxed under these conditions. Thus after the depuration phase, lack of food led particles to persist in digestive tubes. According to these results, intake of silica particles had no retention effects when a food flux was provided, but particles were not easily excreted when the food flux was interrupted.

Caenorhabditis elegans to Model the Capacity of Ascorbic Acid to Reduce Acute Nitrite Toxicity under Different Feed Conditions: Multivariate Analytics on Behavioral Imaging.

Nitrocompounds are present in the environment and human diet and form part of vegetables and processed meat products as additives. These compounds are related to negative impacts on human and animal health. The protective effect of ascorbic acid has been demonstrated by some biological systems as regards several nitrocompounds. This work focused on studying the possibility of modeling this effect on nitrite toxicity with the model Caenorhabditis elegans. The three factors studied in this work were ascorbic acid concentration, nitrite exposure concentration, and presence/absence of food. The protective effect was evaluated by scoring lethality and its impact on behavior by means of multivariate statistical methods and imaging analytics. The effects of nitrite and the influence of food availability were evidenced. Apart from increasing lethality, nitrite had disruption effects on movements. All the observed symptoms reduced when ascorbic acid was administered, and it diminished lethality in all cases. Ascorbic acid maintained nematodes' postural capacities. The results suggest that nitrites' nonspecific toxicity in C. elegans can be mitigated by ascorbic acid, as previously evidenced in other biological systems. Thus, our results reveal the ability of C. elegans to reproduce the known protective effect of ascorbic acid against nitrite.

Toxicological implications of amplifying the antibacterial activity of gallic acid by immobilisation on silica particles: A study on C. elegans.

Immobilisation of natural compounds on solid supports to amplify antimicrobial properties has reported successful results, but modifications to physico-chemical properties can also imply modifications from a toxicological viewpoint. This work aimed to study the immobilising process of gallic acid in the antibacterial activity of L. innocua and its toxicological properties in vivo using Caenorhabditis elegans. The experiment was based on obtaining the minimum bactericidal concentration for free and immobilised gallic acid by comparing lethality, locomotion behaviour, chemotaxis and thermal stress resistance on C.elegans at those concentrations. The results showed a lowering minimum bactericidal concentration and modifications to nematode responses. Increased lethality and velocity of movements was observed. Immobilisation increased the repellent effect of gallic acid with a negative chemotaxis index. Thermal stress resistance was also affected, with higher mortality for immobilised gallic acid compared to bare particles and free gallic acid. Thus despite evidencing a generalised increase in the toxicity of gallic acid in vivo, lowering the minimum bactericidal concentration allowed a bacterial reduction of 99 % with less than one third of mortality for the nematodes exposed to free gallic acid.

Non destructive monitoring of the yoghurt fermentation phase by an image analysis of laser-diffraction patterns: Characterization of cow's, goat's and sheep's milk.

Monitoring yogurt fermentation by the image analysis of diffraction patterns generated by the laser-milk interaction was explored. Cow's, goat's and sheep's milks were tested. Destructive physico-chemical analyses were done after capturing images during the processes to study the relationships between data blocks. Information from images was explored by applying a spectral phasor from which regions of interest were determined in each image channel. The histograms of frequencies from each region were extracted, which showed evolution according to textural modifications. Examining the image data by multivariate analyses allowed us to know that the captured variance from the diffraction patterns affected both milk type and texture changes. When regression studies were performed to model the physico-chemical parameters, satisfactory quantifications were obtained (from R2 = 0.82 to 0.99) for each milk type and for a hybrid model that included them all. This proved that the studied patterns had a common fraction of variance during this processing, independently of milk type.

Effect of tiger-nut (Cyperus esculentus) milk co-product on the surface and diffusional properties of a wheat-based matrix.

The food processing industry generates huge volumes of waste and co-products which still contain valuable compounds. Tiger-nut milk production generates large amounts of a co-product with a high insoluble fibre content, which is interesting as a bioactive component from a nutritional viewpoint. This co-product is formed by two different tissues in composition, particle size and colour terms, so two different flours were obtained from them. Both flours were included in a wheat-based matrix at different substitution levels: 5%, 10% and 20% (d.b). The surface tension of matrices, and the wettability and diffusion of water and oil, were studied. The results showed the matrix's reduced capacity to interact with solvents, principally from the 10% substitution level, with diminished surface tension, and a longer time was needed for both water and oil to wet and diffuse.