Functional properties of a newly cloned fish ortholog of the neutral amino acid transporter B0AT1 (SLC6A19)

The functional properties of an ortholog of the B0AT1 (SLC6A19) amino acid transporter, cloned from the intestine of the sea bass Dicentrachus labrax, were investigated. The two-electrode voltage-clamp technique was applied to Xenopus laevis oocytes heterologously expressing the transporter in order to measure the currents associated with the transport process in different conditions. In particular the substrate specificity, the ionic requirements, and possible effects of pH were examined. Among the organic substrates, leucine, glycine, serine and valine generated the largest transport currents with apparent affinities in the lower millimolar range. The importance of Na+ as the driver ion in the transport process is confirmed, although Li+ is also capable to sustain transport, while K+ is not. No evidence of a relevant role of Cl- in the transport activity was found. Concerning the other two kinds of currents commonly found in electrogenic transporters, very fast presteady-state currents were detected in the absence of organic substrate, while lithium-specific leak currents were not observed. The comparison of these properties with those of the mammalian and insect orthologs may give interesting indication for future structure-function studies in this transporter subfamily

Organic, inorganic and nanoparticles of Se, Zn and Mn in early weaning diets for gilthead seabream (Sparus aurata; Linnaeus, 1758)

Levels of the oxidative stress-related minerals selenium (Se), zinc (Zn) and manganese (Mn) that should be supplied in microdiets for marine fish larvae depend on the availability of the molecular form of these minerals. The objective of this study was to determine how effectively Se, Zn and Mn in organic, inorganic and nanoparticle forms promote larval performance and bone development. Microdiets supplemented with Se, Zn and Mn were fed for 24 days to 20 dah seabream larvae. Microdiets without Se, Zn and Mn supplementation were associated with poor growth, low bone mineralization and a high incidence of anomalies in the branchial arches. Including Zn, Mn and Se in an amino acid chelate organic form promoted maximum larval growth, increased body lipid reserves, enhanced early mineralization and prevented branchial arches anomalies. In contrast, feeding with inorganic forms of these minerals was less effective than organic minerals in improving larval weight or bone mineralization in comparison to the non-supplemented diet. Moreover, the larvae were less resistant to stress, and fish showed higher bone anomalies in the pre-hemal region. Adding Zn, Mn and Se in the form of nanometals did not enhance growth, but improved stress resistance and bone mineralization. The study showed the need to supplement seabream with early weaning diets based on squid meal and krill oil with one or more of the antioxidant minerals, to promote larval growth, bone mineralization and prevention of skeleton anomalies, organic minerals being more effective than inorganic forms and nanometals in promoting mineralization and stress resistance

Inflow and outflow water quality control in coastal aquaculture systems: a case study

Coastal water bodies are a particularly heterogeneous resource, typified by high spatial and temporal variability that could influence the aquaculture in coastal zones. However, the development of coastal aquaculture may produce negative impacts on the coastal area by the potential release of nutrients and organic matter that can be a source of pollution in receiving waters. The aim of this paper was to evaluate the performance of constructed wetland in controlling the dynamics of deoxygenating matter (organic matter and ammonia) and eutrophicating matter [organic matter and soluble reactive phosphorus (SRP)] in the waters entering (inflow) and flowing out (outflow) from a coastal aquaculture fish farm. We observed that constructed wetland systems are effective in removing fractions of total suspended solids, COD, total ammonia nitrogen and SRP contained in the inflow water with higher efficiency in the spring period (60.37%, 14.89%, 65.38% and 17.6% respectively) than in the summer period (45.10%, 8.06%, 32.43% and 8.00% respectively). Similar pattern was recorded for the treatment of the outflow waters, showing that the wetland system reduced most of the deoxygenating and eutrophicating matter produced as a consequence of feeding and fish metabolic activity. During the summer season, high algae mortality can reduce the performance of the wetland system in the outflow water control; this lower efficiency could be improved by controlling the biomass of algae by vegetation harvesting

A Meramod® model approach for the Environmental Impact Assessment (EIA) of the off–shore aquaculture improvement in the Alghero Bay (North western Sardinia, Italy)

Marine fish farming generates particulate wastes which are dispersed in the sea environment. To deal with this problem, particulate waste dispersion models have been developed to predict the effects of fish cage culture. In this study, we evaluated the seabed deposition of a fish farming facility located in the central western Mediterranean by using the Meramod® model. The objectives where first to assess the actual scenario, and second to forecast the possible impact due to the forthcoming enlargement of the farming area with the addition of new fish cages. By computing the hydrodynamic measurements and the daily amount of feed recorded between July and December 2006, the impact seabed surfaces forecasted by the model increased from 5.6ha in the actual scenario, up to 7.3ha in the future. The model estimated a maximum level of total solid flux deposition of 3,800g/m2bed/year and a maximum level of total carbon flux deposition of 1,350g/m2bed/year for both scenarios. Furthermore, the model predicted that the installation of 4 new fish cages (with an hypothetical mean daily amount of feed of 50kg/cage) will produce a total solid and carbon flux deposition levels ranging 0-400 and 0-150g/m2bed/year respectively, under the new fish cages location

A Meramod^® model approach for the Environmental Impact Assessment (EIA) of the off-shore aquaculture improvement in the Alghero Bay (North western Sardinia, Italy)

Marine fish farming generates particulate wastes which are dispersed in the sea environment. To deal with this problem, particulate waste dispersion models have been developed to predict the effects of fish cage culture. In this study, we evaluated the seabed deposition of a fish farming facility located in the central western Mediterranean by using the Meramod® model. The objectives where first to assess the actual scenario, and second to forecast the possible impact due to the forthcoming enlargement of the farming area with the addition of new fish cages. By computing the hydrodynamic measurements and the daily amount of feed recorded between July and December 2006, the impact seabed surfaces forecasted by the model increased from 5.6ha in the actual scenario, up to 7.3ha in the future. The model estimated a maximum level of total solid flux deposition of 3,800g/m2bed/year and a maximum level of total carbon flux deposition of 1,350g/m2bed/year for both scenarios. Furthermore, the model predicted that the installation of 4 new fish cages (with an hypothetical mean daily amount of feed of 50kg/cage) will produce a total solid and carbon flux deposition levels ranging 0-400 and 0-150g/m2bed/year respectively, under the new fish cages location

Neurochemical characterization of myenteric neurons in the juvenile gilthead sea bream (Sparus aurata) intestine

We evaluated the chemical coding of the myenteric plexus in the proximal and distal intestine of gilthead sea bream (Sparus aurata), which represents one of the most farmed fish in the Mediterranean area. The presence of nitric oxide (NO), acetylcholine (ACh), serotonin (5-HT), calcitonin-gene-related peptide (CGRP), substance P (SP) and vasoactive intestinal peptide (VIP) containing neurons, was investigated in intestinal whole mount preparations of the longitudinal muscle with attached the myenteric plexus (LMMP) by means of immunohistochemical fluorescence staining. The main excitatory and inhibitory neurochemicals identified in intestinal smooth muscle were ACh, SP, 5HT, and NO, VIP, CGRP. Some neurons displayed morphological features of ascending and descending interneurons and of putative sensory neurons. The expression of these pathways in the two intestinal regions is largely superimposable, although some differences emerged, which may be relevant to the morphological properties of each region. The most important variances are the higher neuronal density and soma size in the proximal intestine, which may depend on the volume of the target tissue. Since in the fish gut the submucosal plexus is less developed, myenteric neurons substantially innervate also the submucosal and epithelial layers, which display a major thickness and surface in the proximal intestine. In addition, myenteric neurons containing ACh and SP, which mainly represent excitatory motor neurons and interneurons innervating the smooth muscle were more numerous in the distal intestine, possibly to sustain motility in the thicker smooth muscle coat. Overall, this study expands our knowledge of the intrinsic innervation that regulates intestinal secretion, absorption and motility in gilthead sea bream and provides useful background information for rational design of functional feeds aimed at improving fish gut health

Butyrate and taurine exert a mitigating effect on the inflamed distal intestine of European sea bass fed with a high percentage of soybean meal

Abstract Background Due to the paucity of oceanic resources utilized in the preparation of diets for cultured fish, commercial feed producers have been trying to replace fishmeal (FM) using alternative protein sources such as vegetable protein meals (VMs). One of the main drawbacks of using VMs in fish feed is related to the presence of a variety of anti-nutritional factors, which could trigger an inflammation process in the distal intestine. This reduces the capacity of the enterocytes to absorb nutrients leading to reduced fish growth performances. Methods We evaluated the mitigating effects of butyrate and taurine used as feed additives on the morphological abnormalities caused by a soybean meal (SBM)-based diet in the distal intestine of sea bass (Dicentrarchus labrax). We used three experimental diets, containing the same low percentage of FM and high percentage of SBM; two diets were supplemented with either 0.2% sodium butyrate or taurine. Histological changes in the intestine of fish were determined by light and transmission electron microscopy. Infiltration of CD45+ leucocytes in the lamina propria and in the submucosa was assessed by immunohistochemistry. We also quantified by One-Step Taqman® real-time RT-PCR the messenger RNA (mRNA) abundance of a panel of genes involved in the intestinal mucosa inflammatory response such as TNFα (tumor necrosis factor alpha) and interleukins: IL-8, IL-1β, IL-10, and IL-6. Results Fish that received for 2 months the diet with 30% soy protein (16.7% SBM and 12.8% full-fat soy) developed an inflammation in the distal intestine, as confirmed by histological and immunohistochemistry data. The expression of target genes in the intestine was deeply influenced by the type of fish diet. Fish fed with taurine-supplemented diet displayed the lowest number of mRNA copies of IL-1β, IL-8, and IL-10 genes in comparison to fish fed with control or butyrate-supplemented diets. Dietary butyrate caused an upregulation of the TNFα gene transcription. Among the quantified interleukins, IL-6 was the only one to be not influenced by the diet. Conclusions Histological and gene expression data suggest that butyrate and taurine could have a role in normalizing the intestinal abnormalities caused by the SBM, but the underling mechanisms of action seem different

Effect of a specific composition of short- and medium-chain fatty acid 1-Monoglycerides on growth performances and gut microbiota of gilthead sea bream (Sparus aurata)

In aquaculture research, one important aim of gut microbiota studies is to provide the scientific basis for developing effective strategies to manipulate gut microbial communities through the diet, promoting fish health and improving productivity. Currently, there is an increasing commercial and research interest towards the use of organic acids in aquafeeds, due to several beneficial effects they have on growth performance and intestinal tract’s health of farmed fish. Among organic acids, monoglycerides of short-chain fatty acids (SCFAs) and medium-chain fatty acids (MCFAs) have attracted particular research attention also for their bacteriostatic and bactericidal properties. Accordingly, the present study aimed to evaluate the potential beneficial effects of SCFA and MCFA monoglycerides, used as a feed additive, on fish growth performance, and intestinal microbiota composition. For this purpose, a specific combination of short- and medium-chain 1-monoglycerides (SILOhealth 108Z) was tested in 600 juvenile gilthead sea bream (Sparus aurata) of about 60 g mean initial weight that were fed for 90 days with plant-based diets. Two isoproteic and isolipidic diets were formulated. The control fish group received a plant-based diet, whereas the other group received the same control feed, but supplemented with 0.5% of SILOhealth 108Z. The Illumina MiSeq platform for high-throughput amplicon sequencing of 16S rRNA gene and QIIME pipeline were used to analyse and characterize the whole microbiome associated both to feeds and S. aurata intestine. The number of reads taxonomically classified according to the Greengenes database was 394,611. We identified 259 OTUs at 97% identity in sea bream fecal samples; 90 OTUs constituted the core gut microbiota. Firmicutes, Proteobacteria and Actinobacteria represented the dominant phyla in both experimental groups. Among them, relative abundance of Firmicutes and Proteobacteria were positively and negatively affected by dietary SCFA monoglycerides supplementation, respectively. In summary, our findings clearly indicated that SILOhealth 108Z positively modulated the fish intestinal microbiota by increasing the number of beneficial lactic acid bacteria, namely, Lactobacillus, and reducing Gammaproteobacteria, which include several potential pathogenic bacteria. The specific composition of 1-monoglycerides of short- and medium-chain fatty acids contained in SILOhealth 108Z could thus have a great potential as a feed additive in aquaculture