Loss of Ostracoda biodiversity in Western Mediterranean wetlands

• 1. The coastal area of Valencia (Spain) in the Western Mediterranean is rich in freshwater and brackish wetlands. These areas are mostly protected for bird conservation but are also highly affected by human impacts. Six shallow lakes located in Ramsar wetland sites of this area were studied in order to assess past and present ostracod biodiversity and relate it to the status of the lakes, taking into consideration the indicator value of these easily fossilizable crustaceans.
• 2. In all the wetlands studied, ostracod taphocoenoses showed much higher species richness than the corresponding biocoenoses. This is expected from the time accumulative character of the taphocoenosis, but the trend is clearly evident in the most disturbed sites which indicated loss of ostracod diversity owing to human impacts.
• 3. By considering the ecology of most frequent species, multivariate ordination of both tapho- and biocoenoses showed trends within and between lakes in their limnological status, which were related mainly to water chemistry, and also to temporality and eutrophication to a certain extent. Some lakes have apparently lost most of their past ostracod community owing to an increasing trophic status in some cases and to modifications of the hydrological regime in others.

Copyright © 2007 John Wiley & Sons, Ltd.     

Spatial assessment of benefits of a coastal Mediterranean Marine Protected Area

Marine reserves are increasingly advocated not only as conservation but also as fisheries management tools to safeguard the decline of coastal fishing resources. Still, conclusive evidence of their functioning is lacking, amongst others due to the influence of spatio-temporal variations in fish populations and habitat heterogeneity which could hamper a sound data interpretation. We conducted a spatial analysis of the benefits of the Medes Island Marine Reserve by combining geostatistical and Geographic Information System (GIS) tools. Concurrently, we analysed effects of trends reflecting habitat heterogeneity and spatial structuring of data on spatial predictions of fish catch per unit effort (CPUE) and length. Predicted spatial patterns showed the complexity and simultaneous action of trend factors leading to mostly non-linear gradients in CPUE and length data. CPUE of total fish and CPUE and length of common pandora (Pagellus erythrinus) increased close to the Integral Reserve due to direct and indirect reserve effects. CPUE and length of striped red mullet (Mullus surmuletus) slightly increased also near the Integral Reserve, but distinct reserve effects could not be identified due to the strong influence of artificial reefs. We conclude that the spatial dimension of the Buffer Zone, where artisanal fisheries are allowed, permits in general protection only for target species, favouring a habitat with no discontinuities from the reserve outwards. Our spatial approach to assess reserve benefits provides major insights into complex systems like coastal marine reserves in the northwestern Mediterranean. In addition, it contributes to a crucial aspect of marine conservation, viz. the decision on the spatial dimension of protected areas.     

Antigenotoxic effect of grape seed procyanidin extract in Fao cells submitted to oxidative stress

The protective effects of grape seed procyanidin extract on the repair of H(2)O(2)-induced DNA lesions were tested using Fao cells. Cells were exposed to 600 microM H(2)O(2) for 3 or 21 h. A procyanidin extract from grape seed (PE) was incubated or preincubated (1 h) during the exposure to H(2)O(2). The ability of procyanidins to protect against the genotoxicity of H(2)O(2) was compared with those of the monomeric flavanols (+)-catechin and (-)-epicatechin and the flavonol quercetin. After treatment, DNA damage was monitored using alkaline single-cell gel electrophoresis (the comet assay) (Aherne, S. A.; O'Brien, N. M. Nutr. Cancer 1999, 34, 160-166). At the end of the experiment, PE significantly decreased the damage caused by H(2)O(2). The results also showed that quercetin was the most effective of the flavonoids tested, which is consistent with its powerful antioxidant character. The results indicate that procyanidins are more effective than the corresponding individual monomers, catechin and epicatechin, at preventing DNA lesions in hepatocytes and that this protection is higher after preincubation than after co-incubation.     

Bioguided isolation and identification of the nonvolatile antioxidant compounds from fennel (Foeniculum vulgare Mill.) waste

A bioguided isolation of an aqueous extract of fennel waste led to the isolation of 12 major phenolic compounds. Liquid chromatography coupled to atmospheric pressure chemical ionization mass spectrometry (LC/UV/APCI-MS) combined with spectroscopic methods (NMR) was used for compound identification. Radical scavenging activity was tested using three methods: DPPH*, superoxide nitro-blue tetrazolium hypoxanthine/xanthine oxidase, and *OH/luminol chemiluminescence. In addition to products described in the literature, eight antioxidant compounds were isolated and identified for the first time in fennel: 3-caffeoylquinic acid, 4-caffeoylquinic acid, 1,5-O-dicaffeoylquinic acid, rosmarinic acid, eriodictyol-7-O-rutinoside, quercetin-3-O-galactoside, kaempferol-3-O-rutinoside, and kaempferol-3-O-glucoside. The structures of eriodictyol-7-O-rutinoside and quercetin-3-O-glucuronide were completely elucidated by two-dimensional NMR experiments. The isolated compounds exhibited a strong antiradical scavenging activity, which may contribute to the interpretation of the pharmacological effects of fennel.     

Prediction of nutrient digestibility in grower-finisher pigs based on faecal microbiota composition

Microbiota play an important role in total tract nutrient digestion, especially when fibrous diets are fed to pigs. This study aimed to use metagenomics to predict faecal nutrient digestibility in grower-finisher pigs. The study design consisted of 160 three-way crossbreed grower-finisher pigs (80 female and 80 male) which were either fed a diet based on corn/soybean meal or a more fibrous diet based on wheat/barley/by-products. On the day before slaughter, faecal samples were collected and used to determine faecal digestibility of dry matter, ash, organic matter, crude protein, crude fat, crude fibre and non-starch polysaccharides. The faecal samples were also sequenced for the 16S hypervariable region of bacteria (V3/V4) to profile the faecal microbiome. With these data, we calculated the between-animal variation in faecal nutrient digestibility associated with variation in the faecal microbiome, that is the “microbiability”. The microbiability values were significantly greater than zero for dry matter, organic matter, crude protein, crude fibre and non-starch polysaccharides, ranging from 0.58 to 0.93, as well as for crude fat with a value of 0.37, but not significantly different from zero for ash. Using leave-one-out cross-validation, we estimated the accuracy of predicting digestibility values of individual pigs based on their faecal microbiota composition. The accuracies of prediction for crude fat and ash digestibility were virtually 0, and for the other nutrients, the accuracies ranged from 0.42 to 0.63. In conclusion, the faecal microbiota composition gave high microbiability values for faecal digestibility of dry matter, organic matter, crude protein, crude fibre and non-starch polysaccharides. The accuracies of prediction are relatively low if the interest is in precisely predicting faecal nutrient digestibility of individual pigs, but are promising from the perspective of ranking animals in a genetic selection context.     

Bacterial load reduction of live prey for fish larval feeding using Ox-Aquaculture©

Reduction of the bacterial load in live prey is paramount to achieving efficient microbiological control during fish larval rearing. Several methods have been tested using physical and chemical disinfection procedures. Nevertheless, chemicals are difficult to deal with because of the frequent problems encountered when disposing of their residual products and because of the sensitivity of the live prey. Hydrogen peroxide is a good disinfectant that has been used for a long time in other fields of research and decomposes in non‐toxic products. However, it has not been applied for disinfection in aquaculture until recent years. The effects of a hydrogen peroxide‐based product, Ox‐Aquaculture^©, on rotifers and Artemia nauplii survival and on their associated microbial population have been tested in the present study, a disinfection protocol is proposed, and its effects on live prey fatty acids composition after enrichment were checked. More than 80% survival and a 90% reduction of total heterotrophic bacteria and Vibrionaceae were obtained in rotifers exposed for 15 min to 40 mg L^?1 of the product. In the case of Artemia nauplii, 90% survival and 94.5% reduction of heterotrophic bacteria were obtained after 5 min exposure to 8000 mg L^?1 with a further 82.8% reduction of Vibrionaceae. No differences in the fatty acid composition of the live prey were detected after disinfection with hydrogen peroxide. No oxidation of polyunsaturated fatty acids was observed. A short time exposure and easy removal of the product from the treated live prey make hydrogen peroxide an interesting chemical for industrial application.     

Induction of triploidy in the turbot (Scophthalmus maximus): II. Effects of cold shock timing and induction of triploidy in a large volume of eggs

Triploidy was induced in the turbot (Scophthalmus maximus, L.) by applying cold shocks shortly after fertilization. The combined effects of the timing of cold shock commencement after fertilization, cold shock duration and cold shock temperature were investigated. Ploidy was assessed by counting the number of nucleoli per nucleus (NOR) in larvae and also by measuring erythrocyte size in juveniles. A clear peak in triploidy induction was obtained when shocks were started between 6 and 7 min after fertilization at a pre-shock temperature of 13–14°C. With this timing, shocks of 20-min duration at 0°C gave >90% triploidy, with survival about 80% of the untreated controls. In order to ensure both high triploidy rates and high survival, it was necessary to carefully maintain the water temperature just below 0°C. Experiments with small and large volumes of eggs were performed in order to determine how changes in the relative volumes of eggs and chilled water could affect survival and triploidy induction. The best combination to induce triploidy in the turbot was as follows: shock commencement 6.5 min after fertilization, shock duration 25 min, and shock temperature between 0 and −1°C. With this combination, 100% triploidy could consistently be induced with survival 60% of the untreated control. This was successfully applied to a large volume of eggs (300 ml; 1 ml 800 eggs) in order to mass-produce triploid turbot. Triploids had lower survival rate than diploids at hatching but similar thereafter, with the ability to complete the different stages of larval rearing, indicating the viability to produce triploid turbot under farming conditions.     

Stimulatory and inhibitory regulation of DNA synthesis during spermatogenesis: studies in Squalus acanthias

Spermatogenesis is a unique developmental sequence involving multiple cell to cell interactions and several categories of regulatory molecules. In contrast to conventional mammalian models in which testicular organization is highly complex, the testis of the dogfish shark Squalus acanthias is technically advantageous for elucidating stage-dependent structural and functional charactericsics and for in vitro regulatory studies. Using incorporation of [³H]thymidine into acid-insoluble molecules as a measure of DNA synthesis by spermatocysts (germ cell/Sertoli cell units) of premeiotic stages, we obtained evidence of a growth inhibitory bioactivity (chalone) within the testis. This activity is differentially distributed (postmeiotic > meiotic > premeiotic), suggesting that more advanced developmental stages, which are upstream in the vascular pathway within the testis, may control the size of the proliferating spermatocyst population and, hence, the advance of less mature stages. These data provide direct evidence for humoral communication between stages of spermatogenesis.     

Uptake and clearance of exogenous estradiol-17β and testosterone during the early development of coho salmon (Oncorhynchus kisutch), including eggs,alevins and fry

The uptake and clearance of estradiol-17β (E₂) and testosterone (T) were examined during the initial stages of development of coho salmon (Oncorhynchus kisutch), including eyed-eggs, newly hatched alevins and first feeding fry. Radiolabeled steroids were administered through the water in tracer amounts with or without their nonradioactive form at 400 μg l⁻¹. Regardless of developmental stage, saturation levels were invariably attained earlier for T than for E₂, thus resulting in a higher incorporation of E₂. However, both steroids had similar clearance patterns. Uptake and clearance was clearly stage-dependent, being fastest in fry, intermediate in alevins and slowest in eggs. Furthermore, combined uptake and clearance patterns showed that exposure to steroid was also higher for E₂ than for T and stage-dependent, but always markedly highest in alevins. Subsequently, based on the observed elimination of the estrogen, a double immersion in E₂ at 400 μg 1⁻¹, administered 2 days apart to maximize exposure during the alevin stage, was assayed for its effect on sex reversal and found to induce the production of 100% females. We suggest that the yolk, which is present in substantial amounts during the initial stages of development in salmonids, can retain the exogenously administered liposoluble steroids, thus providing developing embryos with an extended supply of, and exposure to, these steroids well after the treatment is finished. Together, these findings help to explain the previously observed high effectiveness of sex steroids administered during early development in regulating gonadal differentiation in salmonids, the higher effectiveness of E₂ compared to T, and clarify the localization of the most sensitive period to the action of exogenous steroids at the alevin stage in the coho salmon.     

Analysis of ochratoxin a in coffee by solid-phase cleanup and narrow-bore liquid chromatography-fluorescence detector-mass spectrometry

A method for the analysis of ochratoxin A (OTA) in green and roasted coffee has been developed. OTA was extracted from coffee with 1% NaHCO(3), and the extract was filtered and purified by solid-phase cleanup using a polymeric column that exhibits reversed-phase and anion-exchange functionalities. OTA was analyzed on a narrow-bore reversed-phase C(18) HPLC column with acetonitrile/water (0.1% formic acid) (40:60) as mobile phase and quantified with a fluorescence detector. The presence of OTA in coffee was confirmed by single-quadruple mass spectrometry using an electrospray ionization source. The method has been validated, obtaining a recovery of 82.5% and a detection limit of 0.1 ng/g. It has been applied to 20 coffee samples from various countries and different manufacturers with no detection of OTA.