Use of alternative protein sources to fishmeal in aquafeed formulations and their effects on gut microbiota of cultured fish.

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, thus promoting fish health and improving productivity. Accordingly, the present PhD project evaluated the effects of different diets on fish growth performance, and gut microbiota composition. Three dietary trials were conducted, using three different alternative sources to fishmeal (poultry by-products, PBP, insect meal, IM, and autolysed yeast from Saccharomyces cerevisiae, AY) in two teleost species: rainbow trout (Oncorhynchus mykiss), and gilthead sea bream (Sparus aurata). In addition to testing each raw material, graded levels of these were also tested, to estimate the more adequate inclusion rate in fish diet. The products tested are not only a substitute of fishmeal per se (e.g. IM or PBP) but can also act as additives (AY) that may directly contribute in enhancing the fish immune system. The Illumina MiSeq platform for high-throughput amplicon sequencing of 16S rRNA gene and QIIME pipeline were used to characterize the whole microbiome associated both, to feeds and fish intestine. Fish fed on experimental diets grew as well as fish fed on control diets. PBP, AY, and IM in the diet affected positively fish gut microbiota, increasing its richness and diversity and in particular, increasing the abundance of beneficial lactic acid- and butyrate-producing bacteria, which contribute to the global health of the host. The operational taxonomic units identified predominantly mapped to the phyla of Firmicutes, Proteobacteria, Bacteroidetes and Actinobacteria. PBP, AY, and IM thus proved to be a suitable protein source for use in aquafeeds

Development of high performance moisture barrier coating for PET bottles

Polyethylene Terephthalate (PET) is one of the most common polymers for packaging applications, because of its properties and versatility. The limit of the use of PET containers in packaging is its low barrier to gases, with a negative impact on the quality of the product. The project intends to improve water vapor barrier properties of PET commercial containers. Possible technological routes should be amenable to an industrial implementation, sustainable and with no impact on bottle appearance. Coating technology can satisfy all the requirements for the industrial implementation, because high performances can be achieved adding a thin barrier material, minimizing the negative impact on the recycling process. The concept at the basis of the PhD activity is the realization of a coating based on a PET compatible polymer, eventually loaded with impermeable 2D-additives, to apply by spray coating on PET bottles. The design of the coating matrix starts from prevalently amorphous vinyl alcohol polymer (HAVOH), that is a high oxygen barrier polymer. HAVOH is a water-soluble polymer, hence a chemical crosslinking agent is required to achieve the insolubility of the coating for water. Crosslinker concentration and crosslinking conditions are investigated by solubility test. WVTR tests on cross-linked HAVOH coating (35µm thickness) show undetectable WVTR (<0.005 g m-2 day-1), even at high relative humidity percentage. Coatings formula designed can be easy removed from PET washing with caustic solution (1%wt) at 85ºC, according to recycling test protocol. Graphene-based additives (graphene, graphene oxide, reduced graphene oxide) are considered as possible additives for the developed coating, to obtain undetectable WVTR, even with thinner coating. Graphene-based fillers are widely used in literature to improve gas barrier properties of a polymer matrix. They reduce gas solubility, due to insolubility of gas in the nanosheets, and diffusivity, as the gas molecules must move around the impermeable nanosheets to diffuse through the polymer. Compared to graphene and GO, RGO is the potential additive for this application. Developed coating is deposited on real PET bottles, to evaluate optimal spray coating conditions, such as air and liquid flow rate, and drying rate. To increase the wettability of the coating solution on PET, the possibility of combine coating with anti-static solvent is considered

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

Next generation sequencing for gut microbiome characterization in rainbow trout (Oncorhynchus mykiss) fed animal by-product meals as an alternative to fishmeal protein sources

Animal by-product meals from the rendering industry could provide a sustainable and commercially viable alternative to fishmeal (FM) in aquaculture, as they are rich in most essential amino acids and contain important amounts of water-soluble proteins that improve feed digestibility and palatability. Among them, poultry by-product meal (PBM) have given encouraging results in rainbow trout (Oncorhynchus mykiss). However, the introduction of new ingredients in the diet needs to be carefully evaluated since diet is one of the main factors affecting the gut microbiota, which is a complex community that contributes to host metabolism, nutrition, growth, and disease resistance. Accordingly, we investigated the effects of partial replacement of dietary FM with a mix of animal by-product meals and plant proteins on intestinal microbiota composition of rainbow trout in relation to growth and feeding efficiency parameters. We used 1540 trout with an initial mean body weight of 94.6 +/- 14.2 g. Fish were fed for 12 weeks with 7 different feed formulations. The growth data showed that trout fed on diets rich in animal by-product meals grew as well as fish fed on control diet, which was rich in FM (37.3%) and PBM-free. High-throughput 16S rRNA gene amplicon sequencing (MiSeq platform, Illumina) was utilised to study the gut microbial community profile. After discarding Cyanobacteria (class Chloroplast) and mitochondria reads a total of 2,701,274 of reads taxonomically classified, corresponding to a mean of 96,474 +/- 68,056 reads per sample, were obtained. Five thousand three hundred ninety-nine operational taxonomic units (OTUs) were identified, which predominantly mapped to the phyla of Firmicutes, Proteobacteria, Bacteroidetes and Actinobacteria. The ratio between vegetable and animal proteins proved to play a central role in determining microbiome profiles and Firmicutes and Proteobacteria phyla were particularly discriminatory for diet type in trout. Plant ingredients favoured a higher Firmicutes: Proteobacteria ratio than animal proteins. Acceptable abundance of Firmicutes was guaranteed by including at least 25% of vegetable proteins in the diet regardless of animal protein source and percentage. In summary animal by-product meals, as replacements to FM, gave good results in terms of growth performances and did not induce significant changes in gut microbial richness, thus proving to be a suitable protein source for use in rainbow trout aqua feed

effect of environmental conditions on the durability of polycarbonate for the protection of cultural heritage sites

Polycarbonate is a good material for covering and protecting cultural heritage sites because of its durability, mechanical properties, and transparency. However, polycarbonate degrades under environmental weathering with a significant decrease of physical and mechanical properties and loss of transparency. In this work, the contemporary presence of ultraviolet irradiation and different temperature and moisture conditions have been taken into account to study the environmental degradation of this polymer with regard to its mechanical and optical properties. The photo-oxidation reactions cause a decrease in the molecular weight and the formation of many oxygenated species. The hydrolytic scission, instead, gives rise to a remarkable reduction in the molecular weight. These two different degradation mechanisms do not seem interconnected because at the lowest degradation temperature and high humidity levels, the reduction of the molecular weight is more pronounced than that observed at the highest temperature but at a lower humidity level. Transparency decreases with the degradative processes, but even after severe degradation the loss of transparency is only about 10%. The yellowness index increases during the first stages of degradation, which has been attributed to the fast formation of carbonyl groups due to photo-oxidation

Mechanical, thermomechanical and reprocessing behavior of green composites from biodegradable polymer and wood flour

The rising concerns in terms of environmental protection and the search for more versatile polymer-based materials have led to an increasing interest in the use of polymer composites filled with natural organic fillers (biodegradable and/or coming from renewable resources) as a replacement for traditional mineral inorganic fillers. At the same time, the recycling of polymers is still of fundamental importance in order to optimize the utilization of available resources, reducing the environmental impact related to the life cycle of polymer-based items. Green composites from biopolymer matrix and wood flour were prepared and the investigation focused on several issues, such as the effect of reprocessing on the matrix properties, wood flour loading effects on virgin and reprocessed biopolymer, and wood flour effects on material reprocessability. Tensile, Dynamic-mechanical thermal (DMTA), differential scanning calorimetry (DSC) and creep tests were performed, pointing out that wood flour leads to an improvement of rigidity and creep resistance in comparison to the pristine polymer, without compromising other properties such as the tensile strength. The biopolymer also showed a good resistance to multiple reprocessing; the latter even allowed for improving some properties of the obtained green composites

Genetic background of mitral valve prolapse

Mitral valve prolapse (MVP) has a prevalence of 2-3% among the population. It involves a heterogeneous group of patients with different expressions and according to the phenotype can be further divided into fibroelastic deficiency, which is mainly considered as a degeneration due to aging, and myxomatous disease, frequently associated with familiar clusters. Thus, MVP can be present in syndromic, when part of a well-defined syndrome, and non-syndromic forms. The latter occurs more often. To the second belong both familiar and isolated or sporadic forms. On one hand, among familial forms, although X-linked transmission related t

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

Production of lead metal by molten-salt electrolysis with energy-efficient electrodes

"The U.S. Bureau of Mines investigated electrode designs for electrowinning lead metal from a LiCl-KCl-PbCl2 electrolyte at 450 deg C. The major objective of this investigation was to decrease the energy requirement for lead electrowinning. Electrolysis was performed in a bench-scale cell using several graphite electrode assemblies. Such parameters as electrode spacing, current density, and chlorine gas removal from the anode were investigated in the bench-scale tests. An optimum electrode assembly, termed the sawtooth design, was scaled up and operated at 3,000 and 4,000 A. The sawtooth electrodes operating at 3,000 A produced lead for 0.66 Kw.H/kg. The electrodes were constructed by cutting large triangular grooves in the electrode surfaces. The electrodes were placed together such that the triangular peaks of one electrode projected into the triangular valleys of the other electrode. Chlorine did not build up on the anode with the sawtooth electrodes." - NIOSHTIC-2NIOSHTIC no. 10008548199