Optimisation of trace mineral supplementation in diets for Atlantic salmon smolt with reference to holistic fish performance in terms of growth, health, welfare, and potential environmental impacts.

The aquafeed ingredient inventory is ever changing, from marine to plant based, and recently evolving to incorporate increasing amounts of low trophic, side stream and circular economy based raw materials, each one contributing with variable amounts and qualities of macro- and micronutrients. Meeting the micronutrient requirement of farmed fish for healthy and efficient growth under normal and challenging conditions is of paramount importance. In this study we run a trial based on a 2 × 4 factorial design with three replications for each dietary treatment, where Atlantic salmon smolt were fed one of 8 experimental diets supplemented with either organic or inorganic mineral premixes (copper, iron, manganese, selenium, and zinc) at four dietary inclusion levels. We saw a trend for higher growth rate in the organic mineral groups irrespective of the dietary mineral levels. Mineral digestibility was negatively correlated with increasing mineral supplementation levels for all tested minerals but Se which increased with the increasing supplementation in the inorganic and up to the 2nd inclusion level in the organic mineral groups. Increasing mineral supplementation affected retention efficiency of Zn, Mn, Cu and Fe while mineral source affected only the retention of Se which was higher in the organic mineral groups. Moreover, fish obtained higher EPA and DHA in their body and increased slaughter yield in the organic as compared to the inorganic mineral groups and corroborated that trace mineral inclusion levels play a key role on salmon fillet's technical quality. More effects from different origin and dietary inclusion levels of trace minerals were seen on fillet yield, fillet technical and nutritional quality, bone strength, skin morphology, organ mineralization and midgut transcriptome.

Individual and combined effects of deoxynivalenol (DON) with other Fusarium mycotoxins on rainbow trout (Oncorhynchus mykiss) growth performance and health.

This study assessed whether the toxicological effects of deoxynivalenol (DON) produced by Fusarium graminearum in rainbow trout (Oncorhynchus mykiss) are altered by the co-exposure to a mixture of toxins produced by Fusarium verticillioides (FUmix). This FUmix contained fusaric acid and fumonisin B1, B2 and B3. Four diets were formulated according to a 2 × 2 factorial design: CON-CON; CON-FUmix; DON-CON; and DON-FUmix. Diets with and without DON contained on average 2700 and 0 µg/kg feed, respectively. The sum of the analysed FUmix toxins was 12,700 and 100 µg/kg feed in the diets with and without FUmix, respectively. The experiment consisted of a 6-week restrictive feeding period immediately followed by a 2-week ad libitum feeding period. Growth performance measurements were taken per feeding period. Histopathological measurements in the liver and gastrointestinal tract (pyloric caeca, midgut and hindgut) were assessed at the end of week 1 and week 6 of the restrictive feeding period and at week 8, the last day of the ad libitum feeding period. During both restrictive and ad libitum feeding, the effects of FUmix and DON on growth performance were additive (no interaction effect; p > 0.05). During the restrictive feeding period, exposure to DON (p ≤ 0.001) and FUmix (p ≤ 0.01) inhibited growth and increased feed conversion ratio (FCR). During this period, DON exposure decreased the protein (p ≤ 0.001) and energy retention (p ≤ 0.05) in the trout. During the ad libitum feeding period, FUmix affected HSI (p ≤ 0.01), while DON exposure reduced feed intake (p ≤ 0.001) and growth (p ≤ 0.001) and increased FCR (p ≤ 0.01). In general, for both liver and intestinal tissue measurements, no interaction effects between DON and FUmix were observed. In the liver, histopathological analysis revealed mild alterations, increased necrosis score by DON (p ≤ 0.01), increased glycogen vacuolization by FUmix (p ≤ 0.05) and decreased percentage of pleomorphic nuclei by FUmix (p ≤ 0.01). DON had a minor impact on the intestinal histological measurements. Over time, some of the liver (glycogen vacuolization score, pleomorphic nuclei; p ≤ 0.01) and intestinal measurements (mucosal fold and enterocyte width; p ≤ 0.01) were aggravated in fish fed the FUmix contaminated diets, with the most severe alterations being noted at week 8. Overall, the co-exposure to FUmix and DON gave rise to additive effects but showed no synergistic or antagonistic effects for the combination of DON with other Fusarium mycotoxins.

Autophagy induction by silver nanowires: a new aspect in the biocompatibility assessment of nanocomposite thin films.

Nanomaterials and their enabled products have increasingly been attracting global attention due to their unique physicochemical properties. Among these emerging products, silver nanowire (AgNW)-based thin films are being developed for their promising applications in next generation nanoelectronics and nanodevices. However, serious concerns remain about possible health and safety risks they may pose. Here, we employed a multi-modal systematic biocompatibility assessment of thin films incorporating AgNW. To represent the possible routes of nanomaterial entry during occupational or environmental exposure, we employed four different cell lines of epithelial, endothelial, gastric, and phagocytic origin. Utilizing a cell-based automated image acquisition and analysis procedure in combination with real-time impedance sensing, we observed a low level of cytotoxicity of AgNW, which was dependent on cell type, nanowire lengths, doses and incubation times. Similarly, no major cytotoxic effects were induced by AgNW-containing thin films, as detected by conventional cell viability and imaging assays. However, transmission electron microscopy and Western immunoblotting analysis revealed AgNW-induced autophasosome accumulation together with an upregulation of the autophagy marker protein LC3. Autophagy represents a crucial mechanism in maintaining cellular homeostasis, and our data for the first time demonstrate triggering of such mechanism by AgNW in human phagocytic cells. Finally, atomic force microscopy revealed significant changes in the topology of cells attaching and growing on these films as substrates. Our findings thus emphasize the necessity of comprehensive biohazard assessment of nanomaterials in modern applications and devices and a thorough analysis of risks associated with their possible contact with humans through occupational or environmental exposure.

The dependence of the optoelectrical properties of silver nanowire networks on nanowire length and diameter.

We have characterized the optoelectrical properties of networks of silver nanowires as a function of nanowire dimension by measuring transmittance (T) and sheet resistance (R(s)) for a large number of networks of different thicknesses fabricated from wires of different diameters (D) and lengths (L). We have analysed these data using both bulk-like and percolative models. We find the network DC conductivity to scale linearly with wire length while the optical conductivity is approximately invariant with nanowire length. The ratio of DC to optical conductivity, often taken as a figure of merit for transparent conductors, scales approximately as L/D. Interestingly, the percolative exponent, n, scales empirically as D², while the percolative figure of merit, Π, displays large values at low D. As high T and low R(s) are associated with low n and high Π, these data are consistent with improved optoelectrical performance for networks of low-D wires. We predict that networks of wires with D = 25 nm could give sheet resistance as low as 25 Ω/□ for T = 90%.

Time- and Dose-Dependent Effects of Dietary Deoxynivalenol (DON) in Rainbow Trout (Oncorhynchus mykiss) at Organism and Tissue Level.

This study with juvenile rainbow trout evaluated the effects of dietary exposure to deoxynivalenol (DON) at industrially relevant doses (up to 1.6 mg/kg) on growth performance, the liver, and the gastrointestinal tract. Fifteen groups of 30 fish each were given one of five dietary treatments in triplicate: (1) control diet (CON; DON < 100 µg/kg feed), (2) naturally DON-contaminated diet (ND1) with a DON content of 700 µg/kg in the feed, (3) ND2 with a DON content of 1200 µg/kg feed, (4) a pure DON-contaminated diet (PD1) with 800 µg/kg of DON in the feed, and (5) PD2 with DON at a concentration of 1600 µg/kg in the feed. The feeding trial lasted eight weeks: six weeks of restrictive feeding followed by two weeks of ad libitum feeding. Exposure to DON during restrictive feeding for six weeks did not affect the growth performance of trout but did lead to a reduction in retained protein in fish fed with higher doses of DON in the ND2 and PD2 groups. During the two following weeks of ad libitum feeding, feed intake was similar among all groups, but body weight gain was lower in the ND2 and PD2 groups and feed efficiency was higher in PD2 (week 8). Histopathological assessment revealed liver damage, including altered nuclear characteristics and haemorrhages, in groups fed higher doses of natural DON (ND2) after just one week of restrictive feeding. Liver damage (necrosis and haemorrhage presence in ND2) was alleviated over time (week 6) but was again aggravated after ad libitum exposure (week 8). In contrast, gastrointestinal tract damage was generally mild with only a few histopathological alterations, and the absence of an inflammatory cytokine response was demonstrated by PCR at week 8. In conclusion, ad libitum dietary exposure of rainbow trout to either natural or pure DON resulted in reduced growth (dose-dependent), while restrictive exposure revealed time-dependent effects of natural DON in terms of liver damage.

Deploying an In Vitro Gut Model to Assay the Impact of the Mannan-Oligosaccharide Prebiotic Bio-Mos on the Atlantic Salmon (Salmo salar) Gut Microbiome.

Alpha mannose-oligosaccharide (MOS) prebiotics are widely deployed in animal agriculture as immunomodulators as well as to enhance growth and gut health. Their mode of action is thought to be mediated through their impact on host microbial communities and their associated metabolism. Bio-Mos is a commercially available prebiotic currently used in the agri-feed industry, but studies show contrasting results of its effect on fish performance and feed efficiency. Thus, detailed studies are needed to investigate the effect of MOS supplements on the fish microbiome to enhance our understanding of the link between MOS and gut health. To assess Bio-Mos for potential use as a prebiotic growth promoter in salmonid aquaculture, we have modified an established Atlantic salmon in vitro gut model, SalmoSim, to evaluate its impact on the host microbial communities. The microbial communities obtained from ceca compartments from four adult farmed salmon were inoculated in biological triplicate reactors in SalmoSim. Prebiotic treatment was supplemented for 20 days, followed by a 6-day washout period. Inclusion of Bio-Mos in the media resulted in a significant increase in formate (P = 0.001), propionate (P = 0.037) and 3-methyl butanoic acid (P = 0.024) levels, correlated with increased abundances of several, principally, anaerobic microbial genera (Fusobacterium, Agarivorans, Pseudoalteromonas). DNA metabarcoding with the 16S rDNA marker confirmed a significant shift in microbial community composition in response to Bio-Mos supplementation with observed increase in lactic acid producing Carnobacterium. In conjunction with previous in vivo studies linking enhanced volatile fatty acid production alongside MOS supplementation to host growth and performance, our data suggest that Bio-Mos may be of value in salmonid production. Furthermore, our data highlights the potential role of in vitro gut models to complementin vivo trials of microbiome modulators. IMPORTANCE In this paper we report the results of the impact of a prebiotic (alpha-MOS supplementation) on microbial communities, using an in vitro simulator of the gut microbial environment of the Atlantic salmon. Our data suggest that Bio-Mos may be of value in salmonid production as it enhances volatile fatty acid production by the microbiota from salmon pyloric ceca and correlates with a significant shift in microbial community composition with observed increase in lactic acid producing Carnobacterium. In conjunction with previous in vivo studies linking enhanced volatile fatty acid production alongside MOS supplementation to host growth and performance, our data suggest that Bio-Mos may be of value in salmonid production. Furthermore, our data highlights the potential role of in vitro gut models to augment in vivo trials of microbiome modulators.

Spray deposition of highly transparent, low-resistance networks of silver nanowires over large areas.

A method to produce scalable, low-resistance, high-transparency, percolating networks of silver nanowires by spray coating is presented. By optimizing the spraying parameters, networks with a sheet resistance of R(s) ≈ 50 Ω â–¡(-1) at a transparency of T = 90% can be produced. The critical processing parameter is shown to be the spraying pressure. Optimizing the pressure reduces the droplet size resulting in more uniform networks. High uniformity leads to a low percolation exponent, which is essential for low-resistance, high-transparency films.

The Occurrence of Mycotoxins in Raw Materials and Fish Feeds in Europe and the Potential Effects of Deoxynivalenol (DON) on the Health and Growth of Farmed Fish Species-A Review.

The first part of this study evaluates the occurrence of mycotoxin patterns in feedstuffs and fish feeds. Results were extrapolated from a large data pool derived from wheat (n = 857), corn (n = 725), soybean meal (n = 139) and fish feed (n = 44) samples in European countries and based on sample analyses by liquid chromatography/tandem mass spectrometry (LC-MS/MS) in the period between 2012-2019. Deoxynivalenol (DON) was readily present in corn (in 47% of the samples) > wheat (41%) > soybean meal (11%), and in aquafeeds (48%). Co-occurrence of mycotoxins was frequently observed in feedstuffs and aquafeed samples. For example, in corn, multi-mycotoxin occurrence was investigated by Spearman's correlations and odd ratios, and both showed co-occurrence of DON with its acetylated forms (3-AcDON, 15-AcDON) as well as with zearalenone (ZEN). The second part of this study summarizes the existing knowledge on the effects of DON on farmed fish species and evaluates the risk of DON exposure in fish, based on data from in vivo studies. A meta-analytical approach aimed to estimate to which extent DON affects feed intake and growth performance in fish. Corn was identified as the ingredient with the highest risk of contamination with DON and its acetylated forms, which often cannot be detected by commonly used rapid detection methods in feed mills. Periodical state-of-the-art mycotoxin analyses are essential to detect the full spectrum of mycotoxins in fish feeds aimed to prevent detrimental effects on farmed fish and subsequent economic losses for fish farmers. Because levels below the stated regulatory limits can reduce feed intake and growth performance, our results show that the risk of DON contamination is underestimated in the aquaculture industry.

SalmoSim: the development of a three-compartment in vitro simulator of the Atlantic salmon GI tract and associated microbial communities.

BACKGROUND: The aquaculture sector now accounts for almost 50% of all fish for human consumption and is anticipated to provide 62% by 2030. Innovative strategies are being sought to improve fish feeds and feed additives to enhance fish performance, welfare, and the environmental sustainability of the aquaculture industry. There is still a lack of knowledge surrounding the importance and functionality of the teleost gut microbiome in fish nutrition. In vitro gut model systems might prove a valuable tool to study the effect of feed, and additives, on the host's microbial communities. Several in vitro gut models targeted at monogastric vertebrates are now in operation. Here, we report the development of an Atlantic salmon gut model, SalmoSim, to simulate three gut compartments (stomach, pyloric caecum, and midgut) and associated microbial communities. RESULTS: The gut model was established in a series of linked bioreactors seeded with biological material derived from farmed adult marine-phase salmon. We first aimed to achieve a stable microbiome composition representative of founding microbial communities derived from Atlantic salmon. Then, in biological triplicate, the response of the in vitro system to two distinct dietary formulations (fishmeal and fishmeal free) was compared to a parallel in vivo trial over 40 days. Metabarcoding based on 16S rDNA sequencing qPCR, ammoniacal nitrogen, and volatile fatty acid measurements were undertaken to survey the microbial community dynamics and function. SalmoSim microbiomes were indistinguishable (p = 0.230) from their founding inocula at 20 days and the most abundant genera (e.g., Psycrobacter, Staphylococcus, Pseudomonas) proliferated within SalmoSim (OTUs accounting for 98% of all reads shared with founding communities). Real salmon and SalmoSim responded similarly to the introduction of novel feed, with majority of the taxa (96% Salmon, 97% SalmoSim) unaffected, while a subset of taxa (e.g., a small fraction of Psychrobacter) was differentially affected across both systems. Consistent with a low impact of the novel feed on microbial fermentative activity, volatile fatty acid profiles were not significantly different in SalmoSim pre- and post-feed switch. CONCLUSION: By establishing stable and representative salmon gut communities, this study represents an important step in the development of an in vitro gut system as a tool for the improvement of fish nutrition and welfare. The steps of the system development described in this paper can be used as guidelines to develop various other systems representing other fish species. These systems, including SalmoSim, aim to be utilised as a prescreening tool for new feed ingredients and additives, as well as being used to study antimicrobial resistance and transfer and fundamental ecological processes that underpin microbiome dynamics and assembly. Video abstract.

Electrical connectivity in single-walled carbon nanotube networks.

Transport in single-walled carbon nanotubes (SWCNTs) networks is shown to be dominated by resistance at network junctions which scale with the size of the interconnecting bundles. Acid treatment, known to dope individual tubes, actually produces a dramatic reduction in junction resistances, whereas annealing significantly increases this resistance. Measured junction resistances for pristine, acid-treated and annealed SWCNT bundles correlate with conductivities of the corresponding films, in excellent agreement with a model in which junctions control the overall network performance.

The effect of dietary n-3/n-6 polyunsaturated fatty acid ratio on salmonid alphavirus subtype 1 (SAV-1) replication in tissues of experimentally infected rainbow trout (Oncorhynchus mykiss).

Salmon pancreas disease (SPD) is one of the most commercially significant viral diseases of farmed Atlantic salmon (Salmo salar) and rainbow trout (Oncorhynchus mykiss) in Europe. In this study, the potential for dietary mitigation of the disease using different polyunsaturated fatty acid (PUFA) profiles was assessed in rainbow trout. We experimentally infected fish with salmonid alphavirus subtype 1 (SAV-1), the causative agent of SPD. These fish were fed two diets with different n-3/n-6 PUFA ratio (high omega 3, 3.08, and high omega 6, 0.87). We assessed the influence of the diets on the fatty acid composition of the heart at 0 days post infection (d.p.i.) (after 4 weeks of feeding the experimental diets prior to SAV-1 infection), and sampled infected and control fish at 5, 15 and 30d.p.i. Viral E1 and E2 glycoprotein genes were quantified by two absolute real-time PCRs in all the organs sampled, and significantly lower levels of the virus were evident in the organs of fish fed with high omega 6. Characteristic pathological lesions were identified in infected fish as early as 5d.p.i., with no significant differences in the pathology lesion scores between the two dietary regimes. This study shows that decreasing the n-3/n-6 PUFA ratio in experimental diets of rainbow trout changes the fatty acid content of the fish, and is associated with reduced SAV-1 replication in rainbow trout.

Size effects and the problem with percolation in nanostructured transparent conductors.

Much research is underway at present to develop nanostructured transparent conductors for use as electrodes. Transparent electrodes typically require high visible transmittances, T > 90%, and so must be very thin. We show that for most nanostructured films thin enough to display T > 90%, the conduction can be described by percolation theory. This means DC conductivities are lower than in bulk, giving correspondingly higher sheet resistances, R(s). To improve our understanding of the consequences of this, we develop a model which relates T to R(s) in the percolation regime. We define a percolative figure of merit, Π, for which high values result in high T and low R(s). High values of Π are achieved for high DC conductivity and low optical conductivity. In addition, the film thickness, t(min), where the DC conductivity first deviates from its bulk value and the percolation exponent, n, must both be as low as possible. We find that this model fits extremely well to much of the data in the literature. We demonstrate that t(min) scales linearly with the smallest dimension of the nanostructure in question (i.e., diameter for wires or thickness for flakes). This clearly confirms that low diameter nanowires or thin platelets are best for transparent conducting applications. We predict the properties of silver nanowire networks to improve as wire diameter is decreased. Networks of wires with D < 20 nm should display properties superior to the best ITO. We demonstrate the deficiencies of standard bulk theory and the importance of understanding percolation by measuring R(s) and T for networks of silver flakes. We measure the bulk ratio of DC to optical conductivity to be ∼35, suggesting R(s) = 100 Ω/◻ and T = 90% are attainable. However, the large flake thickness results in high t(min) and so low Π, resulting in actual values of T = 26% for R(s) = 100 Ω/◻. This makes this material completely unsuitable for transparent conductor applications.

Silver Nanowire Networks as Flexible, Transparent, Conducting Films: Extremely High DC to Optical Conductivity Ratios.

We have used aqueous dispersions of silver nanowires to prepare thin, flexible, transparent, conducting films. The nanowires are of length and diameter close to 6.5 µm and 85 nm, respectively. At low thickness, the films consist of networks but appear to become bulk-like for mean film thicknesses above ∼160 nm. These films can be very transparent with optical transmittance reaching as high as 92% for low thickness. The transmittance (550 nm) decreases with increasing thickness, consistent with an optical conductivity of 6472 S/m. The films are also very uniform; the transmittance varies spatially by typically <2%. The sheet resistance decreases with increasing thickness, falling below 1 Ω/◻ for thicknesses above 300 nm. The DC conductivity increases from 2 × 10(5) S/m for very thin films before saturating at 5 × 10(6) S/m for thicker films. Similarly, the ratio of DC to optical conductivity increases with increasing thickness from 25 for the thinnest films, saturating at ∼500 for thicknesses above ∼160 nm. We believe this is the highest conductivity ratio ever observed for nanostructured films and is matched only by doped metal oxide films. These nanowire films are electromechanically very robust, with all but the thinnest films showing no change in sheet resistance when flexed over >1000 cycles. Such results make these films ideal as replacements for indium tin oxide as transparent electrodes. We have prepared films with optical transmittance and sheet resistance of 85% and 13 Ω/◻, respectively. This is very close to that displayed by commercially available indium tin oxide.

Transparent, flexible, and highly conductive thin films based on polymer-nanotube composites.

We have prepared flexible, transparent, and very conducting thin composite films from poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate), filled with both arc discharge and HIPCO single-walled nanotubes, at high loading level. The films are of high optical uniformity. The arc discharge nanotube-filled composites were significantly more conductive, demonstrating DC conductivities of >10(5) S/m for mass fractions >50 wt %. The ratio of DC to optical conductivity was higher for composites with mass fractions of 55-60 wt % than for nanotube-only films. For an 80 nm thick composite, filled with 60 wt % arc discharge nanotubes, this conductivity ratio was maximized at sigma(DC)/sigma(Op) = 15. This translates into transmittance (550 nm) and sheet resistance of 75 and 80 Omega/square, respectively. These composites were electromechanically very stable, showing <1% resistance change over 130 bend cycles.