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While pesticide use is subject to strict regulatory oversight worldwide, it remains a main concern for environmental protection, including biodiversity conservation. This is partly due to the current regulatory approach that relies on separate assessments for each single pesticide, crop use, and non-target organism group at local scales. Such assessments tend to overlook the combined effects of overall pesticide usage at larger spatial scales. Integrative landscape-based approaches are emerging, enabling the consideration of agricultural management, the environmental characteristics, and the combined effects of pesticides applied in a same or in different crops within an area. These developments offer the opportunity to deliver informative risk predictions relevant for different decision contexts including their connection to larger spatial scales and to combine environmental risks of pesticides, with those from other environmental stressors. We discuss the needs, challenges, opportunities and available tools for implementing landscape-based approaches for prospective and retrospective pesticide Environmental Risk Assessments (ERA). A set of “building blocks” that emerged from the discussions have been integrated into a conceptual framework. The framework includes elements to facilitate its implementation, in particular: flexibility to address the needs of relevant users and stakeholders; means to address the inherent complexity of environmental systems; connections to make use of and integrate data derived from monitoring programs; and options for validation and approaches to facilitate future use in a regulatory context. The conceptual model can be applied to existing ERA methodologies, facilitating its comparability, and highlighting interoperability drivers at landscape level. The benefits of landscape-based pesticide ERA extend beyond regulation. Linking and validating risk predictions with relevant environmental impacts under a solid science-based approach wil

Atlantic horse mackerel (Trachurus trachurus) fillets were coated by gallic acid and chitosan solutions, including chitosan nanoparticles, and fish quality and shelf life were assessed during chilled storage for 13 days. All chitosan-containing coatings decreased microorganisms’ growth in more than 2 log cycles up to late storage stages; however, those with chitosan nanoparticles resulted to be more effective, probably due to a greater contact surface with fish muscle. Furthermore, the lowest total volatile basic nitrogen (TVB-N) values, as well as pH values below 7, were found for horse mackerel fillets coated by those solutions with chitosan nanoparticles. Additionally, horse mackerel fillets coated by solutions with chitosan nanoparticles also showed the lowest thiobarbituric acid reactive substances (TBARS) values, maybe owing to a more sustained release of gallic acid. In this regard, it is worth noting that, although gallic acid solution showed no antimicrobial activity, this prevented lipid oxidation and, therefore, preserved color and texture during the chilling storage.

This study aimed to microencapsulate protein hydrolysates from stripped weakfish (Cynoscion guatucupa) industrial byproducts produced by Alcalase (HA) and Protamex (HP) by spray drying, using maltodextrin as wall material. The physicochemical characteristics, and in vitro antioxidant and Angiotensin-I converting enzyme-inhibitory activities were evaluated during storage. Both microencapsulated hydrolysates showed spherical shape (~3.6 µm particle diameter), low water activity (<0.155) during storage and reduced hygroscopicity (~30%) compared to the free hydrolysate. Infrared spectroscopy evidenced the maltodextrin-hydrolysate interaction. Based on the in vitro results, nematoid C. elegans in L1 larval stage were treated with free and microencapsulated HP, which demonstrated a protective effect on nematoid exposed to oxidative stress (survival ~ 13% control, 77% free HP, and 85% microencapsulated HP) and improved their growth and reproduction rate. Thus, microencapsulation appears to be a good alternative to maintain hydrolysates stability during storage, showing bioactivity in C. elegans.

Branching morphogenesis helps increase the efficiency of gas and liquid transport in many animal organs. Studies in several model organisms have highlighted the molecular and cellular complexity behind branching morphogenesis. To understand this complexity, computational models have been developed with the goal of identifying the “major rules” that globally explain the branching patterns. These models also guide further experimental exploration of the biological processes that execute and maintain these rules. In this paper we introduce the tracheal gills of mayfly (Ephemeroptera) larvae as a model system to study the generation of branched respiratory patterns. First, we describe the gills of the mayfly Cloeon dipterum, and quantitatively characterize the geometry of its branching trachea. We next extend this characterization to those of related species to generate the morphospace of branching patterns. Then, we show how an algorithm based on the “space colonization” concept (SCA) can generate this branching morphospace via growth towards a hypothetical attractor molecule (M). SCA differs from other branch-generating algorithms in that the geometry generated depends to a great extent on its perception of the “external” space available for branching, uses few rules and, importantly, can be easily translated into a realistic “biological patterning algorithm”. We identified a gene in the C. dipterum genome (Cd-bnl) that is orthologous to the fibroblast growth factor branchless (bnl), which stimulates growth and branching of embryonic trachea in Drosophila. In C. dipterum, this gene is expressed in the gill margins and areas of finer tracheolar branching from thicker trachea. Thus, Cd-bnl may perform the function of M in our model. Finally, we discuss this general mechanism in the context of other branching pattern-generating algorithms.

The influence of different melanosis-inhibiting formulations on sensory parameters, texture, water holding capacity and microbiological properties of frozen deep-water rose shrimp (Parapenaeus longirostris) was evaluated throughout 6 months of storage at −18 °C. Eight formulations were tested: three of them containing 4-hexylresorcinol (4-HR, 0.05, 0.1 and 0.25%, w/v) in combination with organic acids (citric, ascorbic and acetic) and chelating agents (ethylenediaminetetraacetic acid and di-sodium di-hydrogen pyrophosphate); four commercial formulae based on sulphites (alone or accompanied by gluconic acid, chitosan or glucose), and a commercial blend based on 4-HR and NaCl. Non-treated shrimp were used as control. 4-HR-based treatments, as well as the joint use of sulphites and d-gluconic acid, were very effective in preventing melanosis during storage without affecting sensory parameters. All formulations assayed did not alter the water holding capacity of muscle protein during frozen storage. In general terms, shrimp hardness increased after a melanosis-inhibiting blend treatment and remained more or less constant during storage. From the microbiological point of view, experimental formulations with 4-HR reduced the total microbiota after treatment, especially at 0.1% (w/w). After 6 months of frozen storage, for most microbial groups studied counts were lower than the limit of detection, except in shrimp treated with the commercial 4-HR-based formulation, where counts were close to 3 log cfu/g. The melanosis-inhibiting blends assayed were useful to preserve sensory, functional and microbial quality in shrimp after six months of frozen storage.

Two moderately halophilic and psychrotolerant new Mycoplasma species were isolated from common cephalopods. Three strains were isolated in pure culture from two individual European flying squid (Todarodes sagittatus), and two individual octopuses (Octopus vulgaris). The strains showed optimal growth at 25 °C and a salinity of 3% (w/v) NaCl. Molecular analyses revealed that the isolates belonged to two new, but phylogenetically related species, divergent from all previously described Mollicutes, representing the first marine isolates of the class, and also the first Mycoplasma strains for which NaCl requirement has been demonstrated. A genome search against all available marine metagenomes and 16S rRNA gene databases indicated that these two species represent a novel non-free-living marine lineage of Mollicutes, specifically associated with marine animals. Morphology and physiology were compatible with other members of this group, and genomic and phenotypic analyses demonstrated that these organisms represent two novel species of the genus Mycoplasma, for which the names Mycoplasma marinum sp. nov. and Mycoplasma todarodis sp. nov. are proposed; the type strains are PET (DSM 105487T, CIP 111404T) and 5HT (DSM 105,488T, CIP 111405T), respectively.

A shrimp extract (SME) obtained from the mild-acid demineralization treatment of shrimp shells to produce chitosan was collected. It was mainly composed of fat (≈73%), protein (≈19%), and ash (≈9%) and contained considerable amounts of calcium (≈1.9 g/100 g), astaxanthin (≈30 mg/100 g) and unsaturated fatty acids (≈27% MUFA, ≈39% PUFA). The SME was used in combination with chitosan for wrapping raw salmon to produce a ready-to-eat product enriched in calcium. No significant changes in hardness were found, as compared to the unwrapped salmon. Estimated intakes of bioaccessible calcium increased significantly by 3.6-fold, whereas intake of bioaccessible fat was reduced by 15%. SFA were the main fatty acid group reduced (≈80%), whereas MUFA and PUFA were only reduced by ≈20% each. Total viable counts, pseudomonads, enterobacteria, and specific fish spoilers were reduced by 2–4 log CFU/g in wrapped sample during the chilled storage period (19 days).

The combined effect of sage (0.3 and 0.6%) and high pressure processing (HPP) [300 MPa (10 min, 9.9 °C) and 600 MPa (10 min, 10.2 °C)] on the antimicrobial and antioxidant characteristics of beef burgers during prolonged chilled storage (60 days) was analysed. Sage powder showed antioxidant and antimicrobial activities, but the addition of sage powder to burgers had no apparent effect on antimicrobial activity; however, antioxidant activity was detected as measured by TBARS, hexanal and photochemiluminescence (PCL). In general, lipid oxidation increased in all samples during storage. HPP at 600 MPa had no effect on lipid oxidation but caused mesophilic and psychrotrophic counts to remain close to the detection limit for at least 6 days. Significant correlations were found between lipid oxidation measured by TBARS and PCL and between TBARS with hexanal over the storage period. Sage had no detrimental effects on sensory attributes of burgers.

The aim of this work was to assess the effect of several melanosis-inhibiting formulations on quality of deepwater pink shrimp (Parapenaeus longirostris) during storage. The formulations tested were as follows: one containing 4-hexylresorcinol (0.1%) in combination with organic acids (citric, ascorbic and acetic) and chelating agents (ethylenediaminetetraacetic acid [EDTA] and di-sodium di-hydrogen pyrophosphate [PPi]); a commercial formula based on sulphites (3%), and another formulation containing a mixture of chitosan (0.3%) and commercial sulphites. Non-treated shrimp were used as control. The 4-hexylresorcinol-based formulation was more effective than 3% of commercial sulphite solutions in preventing melanosis. This treatment prolonged the sensory acceptability of shrimp by up to six days in ice storage. During chilled storage, the treatment with the 4-hexylresorcinol-based formula inhibited the growth of microorganisms by 1.5–2 log cycles as compared to commercial sulphites, especially HS-producers and pseudomonads. At the end of the experimental period, TVB-N levels were slightly higher in the lot treated with 4-hexylresorcinol (33 mg/100 mg) and reflected greater endogenous enzymatic activity, since the counts of the spoiler microorganisms remained around log 5 cfu/g or even lower. In general, the incorporation of chitosan to sulphites did not improve or produce a marked effect on shrimps during storage. During the sensory analysis, the panellists rated all the lots very similar, although the 4-hexylresorcinol formulation was the only one with a neutral odour at the end of the period. The 4-hexylresorcinol-based formulation induced yellow-green colorations to some extent in the cephalothorax, possibly due to the effect of organic acids and chelants, while the rest of the lots presented greyish/blackish colorations. Therefore, the formulation based on 4-hexylresorcinol may safely substitute traditional sulphites to improve the quality of fresh shrimp and prol

The efficacy of hyperbaric cold storage for preserving lean fish has been recently demonstrated but, to the best of our knowledge, no data exist for fatty fish. To evaluate the effect of hyperbaric cold storage on the shelf-life and quality of fatty fish, we stored Atlantic mackerel fillets at 5 °C, both at atmospheric pressure and at 50 MPa. After 12 days of hyperbaric storage, no microbial growth was observed. Thus, most counts were similar or slightly lower than those of control samples at day 0, while H2S-producing organisms and sulphite-reducing bacteria reduced under the detection limits. Moreover, no significant lipid degradation was observed. By contrast, increased microbial counts and significant lipid hydrolysis were detected in the samples stored at atmospheric pressure. Moreover, even though the protein profile was significantly altered during hyperbaric storage, most fish-quality indicators (pH, TVB-N, drip loss, water-holding capacity, and firmness after cooking) were better preserved in the mackerel samples stored at 50 MPa. These results clearly prove that hyperbaric cold storage was more efficient than conventional refrigeration for the preservation of Atlantic mackerel fillets.

This study evaluated the effect of protein hydrolysate (PH) and clove essential oil (CEO) on agar film properties and the shelf-life improvement of flounder fillets. Firstly, Argentine croaker muscle protein was alkali-solubilised and recovered to obtain a stable substrate. This substrate was hydrolysed to 20% degree of hydrolysis, by Alcalase (A20) and Protamex (P20). The molecular weight (MW) distribution, amino acid and in vitro antimicrobial activity of these hydrolysates were tested. Agar films were prepared with and without (control) incorporation of PH (A20 hydrolysate) or CEO and characterised by mechanical properties, water vapour permeability, solubility and optical properties. The microbiological, physical and chemical properties of flounder fillets coated with these films were monitored during storage at 5 °C for 15 days. Compared to P20, A20 hydrolysate possessed lower MW, higher hydrophobic amino acid content and the most effective antimicrobial activity against selected pathogens, such as Staphylococcus aureus, Yersinia enterocolitica, Aeromonas hydrophila, Debaryomyces hansenii and Listeria innocua. The presence of PH increased the water solubility, water vapour permeability, elongation at break and yellowness of the films. The CEO films possessed less transparency than PH films and control films, due to the presence of oil in the formulations. Fillets covered with PH and CEO films, especially the latter, presented lower total volatile bases and pH values, and delayed considerably the growth of HS-producing microorganisms compared to the control films. The CEO might be used as a natural biopreservative to extend the flounder shelf-life. However, further studies are necessary, to assess the antimicrobial effect of PH when used as food packaging.

Introducción El glioblastoma es el tumor cerebral más frecuente. A pesar de los avances en su tratamiento, el pronóstico sigue siendo pobre, con una supervivencia media en torno a los 14 meses. Los costes directos, aquellos asociados al diagnóstico y el tratamiento de la enfermedad, han sido descritos ampliamente. Los costes indirectos, aquellos derivados de la pérdida de productividad debido a la enfermedad, han sido descritos en escasas ocasiones. Material y método Realizamos un estudio retrospectivo, incluyendo a los pacientes diagnosticados entre el 1 de enero del 2010 y el 31 de diciembre del 2013 de glioblastoma en el Hospital Universitario Donostia. Recogimos datos demográficos, relativos al tratamiento ofertado y la supervivencia. Calculamos los costes indirectos a través del método del capital humano, obteniendo datos de sujetos comparables según sexo y edad, y de mortalidad de la población general a través del Instituto Nacional de Estadística. Los salarios pasados fueron actualizados a euros de 2015 según la tasa de inflación interanual y los salarios futuros fueron descontados en un 3,5% anual en forma de interés compuesto. Resultados Revisamos a 99 pacientes, 46 mujeres (edad media 63,53 años) y 53 hombres (edad media 59,94 años). En 29 pacientes se realizó una biopsia y en los 70 restantes se realizó una cirugía resectiva. La supervivencia global media fue de 18,092 meses. Los costes indirectos totales fueron de 11.080.762 € (2015). El coste indirecto medio por paciente fue de 111.926 € (2015). Discusión A pesar de que el glioblastoma es un tipo relativamente poco frecuente de tumor, que supone el 4% de todos los tipos de cáncer, su mal pronóstico y sus posibles secuelas generan una mortalidad y morbilidad desproporcionadamente altas. Esto se traduce en unos costes indirectos muy elevados. El clínico debe ser consciente del impacto del glioblastoma en la sociedad y los costes indirectos deben ser tenidos en cuenta en los estudios de coste-efectividad p, Introduction Glioblastoma is the most common primary brain tumour. Despite advances in treatment, its prognosis remains dismal, with a mean survival time of about 14 months. Many articles have addressed direct costs, those associated with the diagnosis and treatment of the disease. Indirect costs, those associated with loss of productivity due to the disease, have seldom been described. Material and method We conducted a retrospective study in patients diagnosed with glioblastoma at Hospital Universitario Donostia between January 1, 2010 and December 31, 2013. We collected demographics, data regarding the treatment received, and survival times. We calculated the indirect costs with the human capital approach, adjusting the mean salaries of comparable individuals by sex and age and obtaining mortality data for the general population from the Spanish National Statistics Institute. Past salaries were updated to 2015 euros according to the annual inflation rate and we applied a discount of 3.5% compounded yearly to future salaries. Results We reviewed the records of 99 patients: 46 women (mean age 63.53) and 53 men (mean age 59.94); 29 patients underwent a biopsy and the remaining 70 underwent excisional surgery. Mean survival was 18.092 months for the whole series. The total indirect cost for the series was €11 080 762 (2015). Mean indirect cost per patient was €111 926 (2015). Discussion Although glioblastoma is a relatively uncommon type of tumour, accounting for only 4% of all cancers, its poor prognosis and potential sequelae generate disproportionately large morbidity and mortality rates which translate to high indirect costs. Clinicians should be aware of the societal impact of glioblastoma and indirect costs should be taken into account when cost effectiveness studies are performed to better illustrate the overall consequences of this disease.

A functional microemulsified concentrate was obtained from an industrial shrimp cooking effluent by using a centrifugal separator, resulting in a new effluent with only 5% of dry matter. The shrimp concentrate (C) showed valuable properties for use as a bioactive ingredient, being rich in lipids (~16%) and proteins (~11% w/w), and exhibiting antioxidant (radical scavenging capacity and reducing power) and antimicrobial properties. Chitosan–gelatin (ChG) biocomposites with and without the shrimp concentrate were prepared and characterized. The incorporation of C provided film-forming dispersions (S-ChGC) and dried films (F-ChGC) with increased antioxidant and antimicrobial activity. The film microstructure changed with the incorporation of the concentrate, which caused an evident plasticizing effect and reduced the tensile strength. The resulting biocomposites containing the shrimp cooking concentrate showed great potential for use in food applications (as food preservation or food design), medical devices or for cosmetic purposes, as wound dressing or film matrix for application on mucous membranes and epithelia with different functionalities.

Eight seafood protein hydrolysates (SPHs) obtained from squid, shrimp and fish gelatin were incorporated as substitutes of peptones in culture media in order to evaluate its effect on survival and metabolic activity (lactic acid, acetic acid and bacteriocins production) of Enterococcus faecalis DM19. The substitution of commercial peptones in culture media by either a shrimp hydrolysate prepared with Protamex, or by squid protein hydrolysates prepared with Esperase or Alkaline protease, stimulated E. faecalis DM19 growth up to 16%. The incorporation of SPHs, mainly from shrimp, in the culture media significantly increased production of lactic and acetic acids in more than 60%. Furthermore, the media containing SPHs stimulated antimicrobial activity by E. faecalis DM19. The inhibitory activity was observed against both Gram-positive and Gram-negative microorganisms, but it was remarkably observed against Listeria monocytogenes. SPHs incorporated in culture media render properties of bio-technological interest, which, together with their low price, make them suitable for industrial use.

Today, extending the shelf-life of fish, while retaining the organoleptic properties of the product, is still a challenge. To compare the effectiveness of conventional and hyperbaric cold storage in preserving fish quality, we stored Cape hake loins at 5 °C, both at atmospheric pressure and at 50 MPa. After 7 days of storage, microbial counts and total volatile basic-nitrogen content in conventionally refrigerated samples exceeded the limits recommended for consumption. By contrast, hyperbaric cold storage maintained these parameters unaltered, although it produced drip losses close to 5% and increased the shear resistance and whiteness of the raw samples by 44% and 9%, respectively. Nevertheless, after cooking, weight losses were less than half of those of the control loins and whiteness differences disappeared. Consequently, the sensorial analysis could only find moderate differences between the samples before and after hyperbaric storage. These results clearly prove that hyperbaric cold storage was more efficient than conventional refrigeration for the preservation of hake loins.

An active shrimp concentrate (SC), obtained from Litopenaeus vannamei cooking juice, was incorporated into a chitosan-gelatin matrix to develop edible packaging covers (coating and film). Both types of packaging, which showed antimicrobial activity against some fish spoilers and pathogenic organisms, were applied to preserve fish sausages during chilled storage. The storage trial showed that the coating increased the lag phase of total viable microorganisms and enterobacteria to 15 and 10 days, respectively, while the film drastically inhibited growth of these groups. No presence of Staphylococcus aureus or lactic acid bacteria was detected in any batches during storage. Total volatile bases and pH values also decreased during storage, the decrease being more pronounced in the film. With regard to physical-chemical properties, the coated sausages exhibited values for moisture, water holding capacity, and shear strength similar to those of the control sausages until about day 25. The coating was imperceptible and could be a promising way of improving the quality of sausages during storage since it delayed microbial growth. The sausages wrapped in the film showed a drop in pH and moisture, as well as texture hardening, and were described as resembling a pickled product. The sausages were stable for at least 42 days, which represents a shelf life increase of at least this time with respect to the control sausages (in commercial plastic casings). The coating and the film had different effects on the physicochemical and microbiological properties of the fish sausage, and therefore on the stability of the product. In addition, the film triggered modifications in the product that resulted in a product different from the original, offering an attractive alternative with regard to product design.

Hyperbaric storage at room temperature (HS-RT) has recently been proposed as a potential alternative to refrigeration for food preservation. However, there are still very few data about the effects of HS-RT conditions on food microbiota during and after storage. To estimate the effect of pressure level and storage time, we stored strawberry juices at 20 °C and 0.1, 25, 50, 100, and 200 MPa for 1, 10, or 15 days. Total aerobic mesophiles, lactic acid bacteria, and yeasts and molds were quantified before and immediately after storage as well as after a 3-day recovery period at atmospheric pressure and at 20 °C. Both pressure level and storage time affected microbial growth during and after HS-RT. In strawberry juice, storage at 25 MPa retarded microbial growth, but total growth inhibition could not be guaranteed, especially for long storage times. Storage pressures of at least 50 MPa were needed to reduce the initial microbial load during storage. After HS-RT at 25 or 50 MPa, microorganisms could quickly recover their cell-proliferating capacity and, therefore, larger pressures are required to increase microbial stability after decompression. Industrial relevance Hyperbaric storage at room temperature (HS-RT) involves very low energy consumption and, therefore, it can provide an interesting opportunity to reduce energy costs during food storage in comparison with other preservation methods such as freezing or refrigeration. HS-RT could be employed in a wide variety of scenarios: food industry, ship or truck transport for long distances, school or hospital kitchens, restaurants, or even at home. Moreover, its application in developing countries, in which the continuous supply of electric energy is difficult, would be especially noteworthy. However, before industrial implementation, much more research is needed to clarify the effects of the storage conditions on the agents that cause food deterioration (mainly microorganisms and enzymes). The current study contributes t

© 2014 Elsevier Ltd. Chitosan prepared through a mild process using 10% NaOH during deacetylation and a protein concentrate (PCc) obtained from shrimp waste was used to develop active films. Film forming solutions were prepared by solubilizing chitosan with lactic acid without plasticizers addition. The structural properties of films, as observed by optical microscopy, Cryo-SEM, DSC and FTIR, were strongly affected by the addition of the PCc. Films with added PCc were much more tensile resistant but less deformable upon perforation. The presence of PCc also improved film barrier to the light. The films showed good antioxidant and antimicrobial properties, enhanced by the incorporation of PCc. The significance of these films lies in the fact that proteins do not usually appear to enhance antimicrobial properties, and therefore this is especially useful for food applications.

© 2015 Elsevier Ltd. An active coating solution composed of chitosan (Ch) and a shrimp protein-lipid concentrate (PCc), both obtained from Litopenaeus vannamei processing wastes, was applied to preserve shrimp during chilled storage. The addition of PCc increased the antioxidant capacity of the Ch coating, yielding a lower-viscosity mixture which, however, was viscous enough to adhere to the shrimp while maintaining its activity. The shrimp storage trial showed that the Ch coatings, especially when enriched with PCc (Ch-PCc), delayed microbial growth, mainly by extending the lag phase. Pseudomonas spp. appeared to be the predominant bacteria in the microbiota. H2S-producing organisms and luminescent colonies were especially sensitive to this active coating, with inhibition greater than 4 and 2 log cycle respectively, while the growth of lactic acid bacteria was not favored. The Ch-PCc coating delayed the onset of melanosis and did not confer any sensorially detectable color, taste or odor. It therefore shows promise as a means to improve the quality of shrimp during cold storage.

Active biodegradable bilayer films based on soy protein isolate, lignin and formaldehyde were developed. These films showed high water resistance and malleability, making them suitable for use in extreme environmental conditions. The presence of lignin in the formulation provided greater protection against light, especially UV. With the addition of 3%w/w citronella essential oil, the films showed good antifungal activity against Fusarium oxysporum pathogen microorganism in bananas. When the bananas were covered by the films during storage, there was a noticeable reduction in total aerobic mesophiles and moulds and yeasts. As the films aged (1, 3, 6 months), in both controlled and environmental conditions, the main active compounds in the citronella essential oil, citronellal and geraniol, were progressively released from the film matrix, citronellal more than geraniol. Over 30 days of soil degradation, the films lost around 30% of weight irrespective of the conditioning time. © 2014 Elsevier Ltd.

This study evaluates the antioxidant properties and ACE inhibition capacities of eight individual flavonoids before and after incubation with Bifidobacterium animalis ssp. lactis LAFTI@B94®. Changes in flavonoid content and the survival of the probiotics, incubated in individual flavonoid solutions as the only source of carbon, were also determined. The glycosylated flavonoids permitted a higher survival rate of the probiotic bacteria with survival percentages above 50% at 48 h of incubation. The incubation of B. animalis B94 with individual catechins would supposedly produce an increase in all the biological activities tested (ABTS, FRAP and ACE inhibition), but the increments were minor or imperceptible in the flavonol solutions. Epigallocatechin, catechin and epicatechin solutions registered the most important changes in ABTS, FRAP and ACE inhibition capacity, in some cases even doubling their biological activities after incubation with the bacteria. These results are encouraging as the incorporation of B. animalis B94 in various polyphenol-rich foods could be a way of improving the antioxidant properties of the foods. In addition, the polyphenol-rich food could act as a matrix for probiotic strains. © 2014 Elsevier Ltd.

A bioactive film composed of agar, incorporating green tea extract and probiotic strains (Lactobacillus paracasei L26 and Bifidobacterium lactis B94) was applied on hake fillets in order to evaluate the effect of the films during 15 days of storage. Hake was previously inoculated with Shewanella putrefaciens and Photobacterium phosphoreum (103-104CFU/g) to simulate a spoilage process. The green tea and/or probiotic film provoked a reduction, particularly of H2S-producing bacteria counts and total viable bacteria throughout the storage period. The probiotic strains added to the film could pass to the fish producing an increment of lactic acid bacterial counts, even in the presence of green tea extract. The effect of the films also caused a decrease in the indexes of fish quality (total volatile basic nitrogen (TVB-N), trimethylamine nitrogen (TMA-N) and pH). The total viable counts, H2S-producing microorganisms and TVB-N were maintained below the limits of acceptability during 15 days for the fillet covered with the green tea+probiotic film, compared to the rest of the samples. Films with green tea and probiotic were able to extend shelf-life of hake at least for a week and increase the beneficial lactic acid bacteria in fish.

Several shrimp active materials, including protein concentrates, protein hydrolysates and a carotenoid extract, were recovered from shrimp ( Litopenaeus vannamei) processing residues (head and exoskeleton) and characterized in terms of chemical composition and antioxidative properties (Ferric ion reducing power and ABTS radical scavenging capacity). Different coating solutions were prepared by solubilizing chitosan, which was obtained from the residual chitinous material, in lactic acid and blending it with the various recovered materials. Chitosan-based solutions were tested for their viscosity and their antioxidative and antimicrobial properties against 26 selected microbial strains. All blend solutions showed higher antioxidant capacity and reduced viscosity as compared to the base chitosan solution. The incorporation of an astaxanthin-rich protein concentrate increased the antimicrobial capacity of the blend solution, but no further effect was observed after extra addition of the carotenoid extract. © 2013 Elsevier Ltd.

The physico-chemical and functional stability of gelatin (G) and gelatin-lignosulphonate (GLS) films stored during 4 weeks at 21 C, (i) in container or (ii) in contact with oil, was examined. Addition of lignosulphonate dramatically increased ABTS radical scavenging and ferric ion reducing capacities, which remained practically unaltered after the storage period. GLS films exhibited reduced elongation at break, irrespective of storage medium, and retained their water resistance. The feasibility of using GLS film to improve the quality of sardine fillets during chilled storage, alone or in combination with high pressure treatment (300 MPa/10 min/7 C), was evaluated. The combined use of GLS film with high pressure reduced microbial growth, total volatile basic compounds (TVB) and thiobarbituric acid reactive substances (TBARS) during chilled storage. No noteworthy high pressure-induced colour changes were observed in the sardine muscle using this treatment alone, although an increase in yellowness due to the combined treatment was detected. Industrial Relevance Addition of lignosulphonate dramatically increased antioxidant properties (ABTS radical scavenging and ferric ion reducing) of gelatin films, which remained practically unaltered during 4 weeks of storage at room temperature. Application of those films confers stability during storage of chilled sardine, especially in combination with high pressure treatment. These novel packaging was promising for fish preservation. © 2013 Elsevier Ltd.

The incorporation of sunflower protein films with clove essential oil and the application of the resulting materials on the preservation of sardine patties were studied. The addition of clove essential oil to formulations based on sunflower protein concentrates allowed to prepare biodegradable and edible films with increased antioxidant properties and important in vitro antimicrobial properties. The presence of clove essential oil modified protein interactions reducing water solubility and glass transition temperature of resulting films, but it did not modify markedly their moisture content, thickness, color, opacity, water vapor permeability and mechanical properties. When applied to the preservation of refrigerated sardine patties these films allowed to retard fish lipidic auto-oxidation and to slightly delay the growth of total mesophiles, proving once again the importance of checking the films functionality in real food systems. © 2013 Elsevier Ltd.

A green tea aqueous extract was prepared and blended at different percentages (2, 4 and 8%) with a commercial fish-skin gelatin in order to provide gelatin films with antioxidant capacity. This green tea extract proved to be an efficient antioxidant at non-cytotoxic concentrations. Gelatin films with green tea extract were subjected to enzymatic digestion with pepsin (gastric digestion) and with pepsin, trypsin and chymotrypsin (gastrointestinal digestion). The gelatin matrix was efficiently hydrolysed during gastrointestinal digestion and protein hydrolysates composed of low molecular weight peptides, regardless the content of green tea extract, were obtained in all the formulations. High percentages of total polyphenols were recovered from the films with green tea extract after gastrointestinal digestion, although a significant degradation of the major catechins of the green tea (EGCG and EGC) was observed. The increase of the content of green tea extract in the film formulation gave an increase in the antioxidant activity released from the film samples after enzymatic digestion. 85-100% of the maximum expected antioxidant activity was recovered after both gastric and gastrointestinal digestion in spite of the degradation observed of EGCG and EGC. © 2013 Elsevier Ltd.

Active biodegradable films based on agar and agar-fish gelatin were developed by the incorporation of green tea aqueous extract to the film forming solution. The effect of the partial replacement of agar by fish skin gelatin as well as the addition of the green tea extract on the physical properties of the resultant films was evaluated. Special attention was given to the release of antioxidant and antimicrobial compounds from the agar film matrices with and without gelatin. Agar-gelatin films were less resistant and more deformable than agar films. The inclusion of green tea extract decreased tensile strength and elongation at break in both agar and agar-gelatin films. Water vapour permeability and water resistance was not affected either by the replacement of agar by gelatin or the addition of green tea extract, but the water solubility noticeably increased in the films containing green tea extract. The presence of gelatin in the agar-green tea matrix film hindered the release of total phenolic compounds, catechins and flavonols in water. As a consequence, the antioxidant power released by the films was lower in the case of films containing gelatin. However, the antimicrobial activity of the films was not affected by the presence of gelatin. © 2012 Elsevier Ltd.

This work deals with the design and formulation of gelatin edible coatings and films of incorporated with probiotic bacteria Lactobacillus acidophilus and Bifidobacterium bifidum. A gelatin coating incorporated with B. bifidum was applied to fish to assess its effect during chilled storage. B. bifidum remained viable during the storage and H2S-producing microorganisms were reduced in 2 log cycles. In a further experiment, fish coated with gelatin films incorporated with bifidobacteria was treated with a high pressure (200 MPa/10 min/20°C). At the end of the storage, a reduction of total viable counts (< 2 log cycles) were obtained. Moreover, the combined treatments drastically reduced the content of volatile bases and lowered the pH by more than 1 unit. Furthermore, the application of gelatin edible packaging incorporated with bacteria can be promising for fish preservation, especially when combined with other technologies such as a high-pressure. Industrial relevance: Gelatin edible coatings and films could act as a proper matrix to incorporate lactic acid bacteria to some foods. The application of those edible packaging confer stability during storage of chilled fish, especially in combination with high pressure treatment. These novel packaging was promising for fish preservation. © 2012 Elsevier Ltd.

The aim of this study was to assess the potential activation of sunflower protein films with antioxidant and antimicrobial properties conferred by the phenolic compounds of sunflower seeds, which remain associated to the proteins used as starting material for film preparation. Two sunflower protein concentrates obtained from the residual pellet of oil industry were used, which had different content of phenolic compounds, mainly chlorogenic and caffeic acids. The film-forming dispersions and the films obtained were analyzed regarding their antioxidant properties (using ABTS, FRAP and PCL assays) and their antimicrobial properties (by agar disk diffusion tests). Phenolic compounds conferred important antioxidant properties to both dispersions and films, this activity being dependent on their content and their free or protein-bound nature. These compounds, however, did not confer their characteristic antimicrobial properties reported in previous studies, possibly due to their interaction with proteins and the pH of the film-dispersions. Since the antioxidant activity of phenolic compounds was preserved during the protein concentration process (inherent to the film formation), these protein matrices may be considered useful for protecting these bioactive compounds. © 2012 Elsevier Ltd.

Simple edible films can be manufactured to meet not only their primary protective purpose but can be easily manipulated to meet sensory expectations and to contain compounds which enhance the protective properties or even have the potential to deliver health benefits. However, the use of such edible films not only to protect the food but as a vehicle to deliver health benefits has not been investigated. In this paper we study agar films containing an aqueous extract of green tea, rich in polyphenol compounds, and the bioaccessibility of these compounds during simulated digestion in the upper gastro-intestinal tract using a dynamic gastric model (DGM) and a static duodenal model. It is concluded that the recovery of the tea compounds added to the agar film mainly occurs in the stomach (50-80%) and that little or no additional recovery is observed in the duodenum. Furthermore, the green tea compounds recovered show both reducing power and radical scavenging ability, but not antimicrobial activity. The bioaccessibility of the green tea flavonols is reduced in the presence of gelatin used to simulate the presence of protein in the stomach, but it is not clear if this is due to reduced release or sequestration of released compounds by the gelatin. © 2012.

The rising interest in the valorisation of industrial by-products is one of the main reasons why exploring different species and optimizing the extracting conditions of collagen and gelatin has attracted the attention of researchers in the last decade. The most abundant sources of gelatin are pig skin, bovine hide and, pork and cattle bones, however, the industrial use of collagen or gelatin obtained from non-mammalian species is growing in importance. The classical food, photographic, cosmetic and pharmaceutical application of gelatin is based mainly on its gel-forming properties. Recently, and especially in the food industry, an increasing number of new applications have been found for gelatin in products such as emulsifiers, foaming agents, colloid stabilizers, biodegradable film-forming materials and micro-encapsulating agents, in line with the growing trend to replace synthetic agents with more natural ones. In the last decade, a large number of studies have dealt with the enzymatic hydrolysis of collagen or gelatin for the production of bioactive peptides. Besides exploring diverse types of bioactivities, of an antimicrobial, antioxidant or antihypertensive nature, studies have also focused on the effect of oral intake in both animal and human models, revealing the excellent absorption and metabolism of Hyp-containing peptides. The present work is a compilation of recent information on collagen and gelatin extraction from new sources, as well as new processing conditions and potential novel or improved applications, many of which are largely based on induced cross-linking, blending with other biopolymers or enzymatic hydrolysis. © 2011 Elsevier Ltd.

Essential oils of clove (Syzygium aromaticum L.), fennel (Foeniculum vulgare Miller), cypress (Cupressus sempervirens L.), lavender (Lavandula angustifolia), thyme (Thymus vulgaris L.), herb-of-the-cross (Verbena officinalis L.), pine (Pinus sylvestris) and rosemary (Rosmarinus officinalis) were tested for their antimicrobial activity on 18 genera of bacteria, which included some important food pathogen and spoilage bacteria. Clove essential oil showed the highest inhibitory effect, followed by rosemary and lavender. In an attempt to evaluate the usefulness of these essential oils as food preservatives, they were also tested on an extract made of fish, where clove and thyme essential oils were the most effective. Then, gelatin-chitosan-based edible films incorporated with clove essential oil were elaborated and their antimicrobial activity tested against six selected microorganisms: Pseudomonas fluorescens, Shewanella putrefaciens, Photobacterium phosphoreum, Listeria innocua, Escherichia coli and Lactobacillus acidophilus. The clove-containing films inhibited all these microorganisms irrespectively of the film matrix or type of microorganism. In a further experiment, when the complex gelatin-chitosan film incorporating clove essential oil was applied to fish during chilled storage, the growth of microorganisms was drastically reduced in gram-negative bacteria, especially enterobacteria, while lactic acid bacteria remained practically constant for much of the storage period. The effect on the microorganisms during this period was in accordance with biochemical indexes of quality, indicating the viability of these films for fish preservation. © 2010 Elsevier Ltd.

This work evaluates the physicochemical and sensorial qualities of salmon, tuna and desalted >bacalao>, thinly sliced as carpaccios, and subjected to 15-min of continuous pressure and pulsed pressure in three 5-min steps (200-300 MPa at 7 °C). Pressurization of salmon and tuna gave rise to an increase in the shear strength of carpaccios, a reduction of the water and lipid binding properties and an increase in the total colour difference. Though these changes were also detected by the sensory panel, the resulting pressurized products obtained high scores for acceptability. Desalted >bacalao> carpaccio was more stable under high pressure and sensory analysis revealed that in most treatments the raw attributes had been retained. The results obtained by applying pressure in one or three consecutive cycles were, in general, the same. High-pressure treatment was shown to be an adequate tool for obtaining high-quality desalted >bacalao> carpaccio, whereas salmon and tuna suffered a loss of raw attributes. However the resulting products acquired new sensory features which were highly accepted by the sensory panel. Industrial relevance: Nowadays, there is a growing interest in consuming raw or minimally processed foods and among fishery products, carpaccio are becoming increasingly popular. The present work deals with the effect of high pressure on the physico-chemical and sensory attributes of carpaccios made from salmon, tuna, and desalted >bacalao>. High pressure may be an adequate tool to obtain high quality carpaccio or carpaccio-like products, as the products acquired new sensory features which were highly accepted by the sensory panel. From the industry viewpoint the application of high pressure to fish carpaccios may be appealing in a two way: firstly, the resulting carpaccio are presumably free of parasites and present a better microbiological quality, thereby increasing the shelf-life. Secondly, products may acquire new sensory attributes that can be very appreciat

Most films used to preserve foodstuffs are made from synthetic plastic materials. However, for environmental reasons, attention has recently turned to biodegradable films. Gelatin has been extensively studied for its film-forming capacity and applicability as an outer covering to protect food against drying, light, and oxygen. Moreover, it is one of the first materials proposed as a carrier of bioactive components. Gelatins from alternatives to mammalian species are gaining prominence, especially gelatins from marine fish species. Because of their good film-forming abilities, fish gelatins may be a good alternative to synthetic plastics for making films to preserve foodstuffs. The mechanical and barrier properties of these films depend largely on the physical and chemical characteristics of the gelatin, especially the amino acid composition, which is highly species specific, and the molecular weight distribution, which depends mainly on processing conditions. Different film formulations can be developed to extend the films' physical and chemical properties and to add new functional attributes. This paper reviews the most recent scientific literature dealing with films based on gelatins from different fish species and considers various strategies intended to improve the physical properties of such films by combining fish gelatins with such other biopolymers as soy protein isolate, oils and fatty acids, and certain polysaccharides. The use of plasticizers and cross-linking agents is also discussed. Specific attributes, such as antimicrobial and antioxidant activities, may be also conferred by blending the gelatin with chitosan, lysozyme, essential oils, plant extracts, or vitamin C to produce an active packaging biomaterial.

A comparison was made of the effects on melanosis development in Norway lobsters (Nephrops norvegicus) of treatment by dusting with a commercial sulphite-based product and of spraying with a formulation containing 4-hexylresorcinol (0.1% and 0.05%), in combination with organic acids and chelating agents. The following tests were performed during chilled storage: polyphenol oxidase (PPO) activity, melanosis score, colour parameters, tyrosine and tyramine content, as the main substrate of PPO. Differences among treatments were evaluated by means of statistical analyses (ANOVA, principal components and discriminant analyses). All formulations diminished PPO activity during storage successfully. The melanosis score was higher in sulphite-treated Norway lobsters, and a formulation with 0.05% 4-hexylresorcinol was enough to prevent the appearance of melanosis for 12 days. The tyrosine content decreased during storage, but the tyramine content was insignificant. Formulations with 4-hexylresorcinol improved the appearance of Norway lobsters, in comparison with the commercial sulphite-based product. © 2005 Elsevier Ltd. All rights reserved.

A coating made in cold from a blend of a chitosan and a gelatin solution was applied to patties made of chilled cod, and its preservative effect was assessed by colour measurements, rheological measurements (hardness, elasticity, cohesiveness, chewiness, gumminess, and adhesiveness), biochemical determinations (total volatile bases and thiobarbituric acid as measures of rancidity) and microbiological assays (total bacterial counts, luminiscent bacteria, enterobacteria, pseudomonas, lactic acid bacteria, and Staphylococcus aureus). The effect of dry powdered chitosan mixed into the patties was tested as well. The use of chitosan either as a coating or as a powdered ingredient did not affect lightness at the end of the storage period considered but did result in an increase in the value of yellowness. The coating increased patty elasticity, whereas adding powdered chitosan to the patty mixture increased the other rheological parameter values. The findings on the effect of the chitosan on rancidity were not conclusive due to the low values recorded in the cod. However, the coating did prevent spoilage of the cod patties as reflected by a decrease in total volatile basic nitrogen and in the microorganism counts, in particular counts of gram-negative bacteria. In contrast, none of these effects on spoilage were observed when the chitosan was added to the patty mixture in powdered form. Coatings prepared in cold from a blend of gelatin and chitosan offer a promising alternative for preserving fish patties.

The microflora of soaked cod products from salted and dried salted cod were identified to the species level by the Vitek® automated identification system using the Gram-positive identification (GPI) and Gram-negative identification plus (GNI+) cards, and a cluster analysis was performed to determine the similarity among the strains. Within the Gram-positive bacteria, Staphylococcus epidermidis, S. hominis, S. warneri, S. haemolyticus, S. simulans and S. saprophyticus were found. Regarding Gram-negative, Stenotrophomonas maltophilia, Vibrio alginolyticus, Pasteurella haemolytica, Enterobacter agglomerans, E. asburiae, E. cloacae, Pseudomonas putida, P. fluorescens, Shewanella putrefaciens, Morganella morganii, Actinobacillus urea, Acinetobacter lwoffi/junii and non-fermentative bacilli were identified. Several species found in the soaked products were also isolated from the salted and dried salted cod. Thus, despite of the low water activity of these products, the mentioned strains were able to survive. Some of these, had the ability to produce H2S and/or presented decarboxylase activity, therefore it is probable that the growth of these bacteria during soaking might contribute to the spoilage of soaked cod. Aeromonas hydrophila and A. caviae were not isolated from the soaked food, but from the raw material (dried salted cod). Their presence might constitute a potential hazard, especially considering that these foods may be eaten raw.

Growth, trimethylamine (TMA), off-odour and biogenic amine production by a strain of Shewanella putrefaciens isolated from spoiled hake (Merluccius merluccius L.) and cultured in a model system, were tested under four different gas compositions (60% CO2/40% O2, 60% CO2/15% O2/25% N2, 40% CO2/60% O2, 40% CO2/40% O2/20% N2) and under air. After 3 weeks of incubation, the control (air) batch showed the highest microbial counts (>9 log cfu/ml) and TMA concentrations (45 mg N-TMA/100 ml), and strong putrid off-odours were detected from day 15. High amounts of putrescine and cadaverine were produced in this batch, but histamine increased only slightly. Batches under controlled atmospheres showed reduced growth, TMA, off-odour and biogenic amine production. The 40% CO2/60% O2 mixture had the strongest inhibitory effect on bacterial growth, while the 60% CO2/15% O2/25% N2 mixture was less effective. Putrescine and histamine production was lowest in S. putrefaciens under the 40% CO2/60% O2 mixture. However, the level of histamine in S. putrefaciens was higher under 40% CO2/40% O2/20% N2 than when the bacteria was incubated in air. Under the gas mixtures, there was a similar decrease in the production of cadaverine and agmatine by S. putrefaciens, irrespective of the gas concentrations. The production of 2-phenylethylamine appeared to be inhibited under any atmospheric condition. Copyright © 2001 Elsevier Science B.V.