| 1. |
- Zeng, Jiao, et al.
(författare)
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High-resolution structure of a fish aquaporin reveals a novel extracellular fold
- 2022
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Ingår i: Life Science Alliance. - : Life Science Alliance, LLC. - 2575-1077. ; 5:12
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Tidskriftsartikel (refereegranskat)abstract
- Aquaporins are protein channels embedded in the lipid bilayer in cells from all organisms on earth that are crucial for water homeostasis. In fish, aquaporins are believed to be important for osmoregulation; however, the molecular mechanism behind this is poorly understood. Here, we present the first structural and functional characterization of a fish aquaporin; cpAQP1aa from the fresh water fish climbing perch (Anabas testudineus), a species that is of high osmoregulatory interest because of its ability to spend time in seawater and on land. These studies show that cpAQP1aa is a water-specific aquaporin with a unique fold on the extracellular side that results in a constriction region. Functional analysis combined with molecular dynamic simulations suggests that phosphorylation at two sites causes structural perturbations in this region that may have implications for channel gating from the extracellular side.
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| 2. |
- Andersson, Marica, et al.
(författare)
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Low Holding Densities Increase Stress Response and Aggression in Zebrafish
- 2022
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Ingår i: Biology. - : MDPI AG. - 2079-7737. ; 11:5
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Tidskriftsartikel (refereegranskat)abstract
- Simple Summary Zebrafish are used as experimental animals in labs all around the world. To ensure that the health of zebrafish is maintained at the highest level, it is important to know the optimal housing conditions of the animals, including the housing density. Guidelines for housing densities of zebrafish can then be spread and followed globally, making it possible to compare research data from different facilities. To investigate the optimal housing densities of zebrafish and to better understand how holding density affects zebrafish behaviour and physiology, we evaluated the welfare of zebrafish housed at different densities for nine weeks. We observed that fish housed at the lowest holding density of 1 fish/L stood out from the rest of the experimental fish, showing higher levels of aggression, secreting more of the stress hormone cortisol in the water, and spending more time in the top zone of the tank, possibly reflecting the fact that fish in this density were hiding more behind the floating plants. Our data indicate that zebrafish should not be kept at 1 fish/L, or lower, to ensure good welfare of the animals. With laboratory zebrafish (Danio rerio) being an established and popular research model, there is a need for universal, research-based husbandry guidelines for this species, since guidelines can help promote good welfare through providing appropriate care. Despite the widespread use of zebrafish in research, it remains unclear how holding densities affect their welfare. Previous studies have mainly evaluated the effects of holding densities on a single parameter, such as growth, reproductive output, or social interactions, rather than looking at multiple welfare parameters simultaneously. Here we investigated how chronic (nine weeks) exposure to five different holding densities (1, 4, 8, 12, and 16 fish/L) affected multiple welfare indicators. We found that fish in the 1 fish/L density treatment had higher free water cortisol concentrations per fish, increased vertical distribution, and displayed aggressive behaviour more frequently than fish held at higher densities. On the other hand, density treatments had no effect on anxiety behaviour, whole-brain neurotransmitter levels, egg volume, or the proportion of fertilised eggs. Our results demonstrate that zebrafish can be held at densities between 4 and 16 fish/L without compromising their welfare. However, housing zebrafish in the density of 1 fish/L increased their stress level and aggressive behaviour.
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| 3. |
- Benktander, John, et al.
(författare)
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Gill Mucus and Gill Mucin O-glycosylation in Healthy and Amebic Gill Disease-Affected Atlantic Salmon.
- 2020
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Ingår i: Microorganisms. - : MDPI AG. - 2076-2607. ; 8:12
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Tidskriftsartikel (refereegranskat)abstract
- Amoebic gill disease (AGD) causes poor performance and death in salmonids. Mucins are mainly comprised by carbohydrates and are main components of the mucus covering the gill. Since glycans regulate pathogen binding and growth, glycosylation changes may affect susceptibility to primary and secondary infections. We investigated gill mucin O-glycosylation from Atlantic salmon with and without AGD using liquid chromatography-mass spectrometry. Gill mucin glycans were larger and more complex, diverse and fucosylated than skin mucins. Confocal microscopy revealed that fucosylated mucus coated sialylated mucus strands in ex vivo gill mucus. Terminal HexNAcs were more abundant among O-glycans from AGD-affected Atlantic salmon, whereas core 1 structures and structures with acidic moieties such as N-acetylneuraminic acid (NeuAc) and sulfate groups were less abundant compared to non-infected fish. The fucosylated and NeuAc-containing O-glycans were inversely proportional, with infected fish on the lower scale of NeuAc abundance and high on fucosylated structures. The fucosylated epitopes were of three types: Fuc-HexNAc-R, Gal-[Fuc-]HexNAc-R and HexNAc-[Fuc-]HexNAc-R. These blood group-like structures could be an avenue to diversify the glycan repertoire to limit infection in the exposed gills. Furthermore, care must be taken when using skin mucus as proxy for gill mucus, as gill mucins are distinctly different from skin mucins.
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| 4. |
- Benktander, John, et al.
(författare)
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Stress impairs skin barrier function and induces α2-3 linked n-acetylneuraminic acid and core 1 o-glycans on skin mucins in atlantic salmon, salmo salar
- 2021
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Ingår i: International Journal of Molecular Sciences. - : MDPI AG. - 1661-6596 .- 1422-0067. ; 22:3
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Tidskriftsartikel (refereegranskat)abstract
- © 2021 by the authors. Licensee MDPI, Basel, Switzerland. The skin barrier consists of mucus, primarily comprising highly glycosylated mucins, and the epithelium. Host mucin glycosylation governs interactions with pathogens and stress is associated with impaired epithelial barrier function. We characterized Atlantic salmon skin barrier function during chronic stress (high density) and mucin O-glycosylation changes in response to acute and chronic stress. Fish held at low (LD: 14–30 kg/m3) and high densities (HD: 50-80 kg/m3) were subjected to acute stress 24 h before sampling at 17 and 21 weeks after start of the experiment. Blood parameters indicated primary and secondary stress responses at both sampling points. At the second sampling, skin barrier function towards molecules was reduced in the HD compared to the LD group (Papp mannitol; p < 0.01). Liquid chromatography–mass spectrometry revealed 81 O-glycan structures from the skin. Fish subjected to both chronic and acute stress had an increased proportion of large O-glycan structures. Overall, four of the O-glycan changes have potential as indicators of stress, especially for the combined chronic and acute stress. Stress thus impairs skin barrier function and induces glycosylation changes, which have potential to both affect interactions with pathogens and serve as stress indicators.
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| 5. |
- Brijs, J., et al.
(författare)
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Prevalence and severity of cardiac abnormalities and arteriosclerosis in farmed rainbow trout (Oncorhynchus mykiss)
- 2020
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Ingår i: Aquaculture. - : Elsevier BV. - 0044-8486 .- 1873-5622. ; 526
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Tidskriftsartikel (refereegranskat)abstract
- Cardiovascular disease may pose a major threat to the health and welfare of farmed fish. By investigating a range of established cardiovascular disease indicators, we aimed to determine the prevalence, severity and consequences of this affliction in farmed rainbow trout (Oncorhynchus mykiss) from an open cage farm in the Baltic Sea, an open cage farm in a freshwater lake, and a land-based recirculating aquaculture system. We also aimed to identify environmental, anthropogenic and physiological factors contributing towards the development of the disease. The majority of trout possessed enlarged hearts with rounded ventricles (mean height:width ratios of 1.0–1.1 c.f. ~1.3 in wild fish) and a high degree of vessel misalignment (mean angles between the longitudinal ventricular axis and the axis of the bulbus arteriosus of 28–31 °c.f. ~23° in wild fish). The prevalence and severity of coronary arteriosclerosis was also high, as 92–100% of fish from the different aquaculture facilities exhibited coronary lesions. Mean lesion incidence and severity indices were 67–95% and 3.1–3.9, respectively, which resulted in mean coronary arterial blockages of 19–32%. To evaluate the functional significance of these findings, we modelled the effects of arterial blockages on coronary blood flow and experimentally tested the effects of coronary occlusion in a sub-sample of fish. The observed coronary blockages were estimated to reduce coronary blood flow by 34–54% while experimental coronary occlusion adversely affected the electrocardiogram of trout. Across a range of environmental (water current, predation), anthropogenic (boat traffic intensity, hatchery of origin, brand of feed pellets) and physiological factors (condition factor, haematological and plasma indices), the hatchery of origin was the main factor contributing towards the observed variation in the development of cardiovascular disease. Therefore, further research on the effects of selective breeding programs and rearing strategies on the development of cardiovascular disease is needed to improve the welfare and health of farmed fish. © 2020 The Authors
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| 6. |
- Clark, M. S., et al.
(författare)
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Deciphering mollusc shell production: the roles of genetic mechanisms through to ecology, aquaculture and biomimetics
- 2020
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Ingår i: Biological Reviews. - : Wiley. - 1464-7931 .- 1469-185X. ; 95:6, s. 1812-37
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Tidskriftsartikel (refereegranskat)abstract
- Most molluscs possess shells, constructed from a vast array of microstructures and architectures. The fully formed shell is composed of calcite or aragonite. These CaCO(3)crystals form complex biocomposites with proteins, which although typically less than 5% of total shell mass, play significant roles in determining shell microstructure. Despite much research effort, large knowledge gaps remain in how molluscs construct and maintain their shells, and how they produce such a great diversity of forms. Here we synthesize results on how shell shape, microstructure, composition and organic content vary among, and within, species in response to numerous biotic and abiotic factors. At the local level, temperature, food supply and predation cues significantly affect shell morphology, whilst salinity has a much stronger influence across latitudes. Moreover, we emphasize how advances in genomic technologies [e.g. restriction site-associated DNA sequencing (RAD-Seq) and epigenetics] allow detailed examinations of whether morphological changes result from phenotypic plasticity or genetic adaptation, or a combination of these. RAD-Seq has already identified single nucleotide polymorphisms associated with temperature and aquaculture practices, whilst epigenetic processes have been shown significantly to modify shell construction to local conditions in, for example, Antarctica and New Zealand. We also synthesize results on the costs of shell construction and explore how these affect energetic trade-offs in animal metabolism. The cellular costs are still debated, with CaCO(3)precipitation estimates ranging from 1-2 J/mg to 17-55 J/mg depending on experimental and environmental conditions. However, organic components are more expensive (similar to 29 J/mg) and recent data indicate transmembrane calcium ion transporters can involve considerable costs. This review emphasizes the role that molecular analyses have played in demonstrating multiple evolutionary origins of biomineralization genes. Although these are characterized by lineage-specific proteins and unique combinations of co-opted genes, a small set of protein domains have been identified as a conserved biomineralization tool box. We further highlight the use of sequence data sets in providing candidate genes forin situlocalization and protein function studies. The former has elucidated gene expression modularity in mantle tissue, improving understanding of the diversity of shell morphology synthesis. RNA interference (RNAi) and clustered regularly interspersed short palindromic repeats - CRISPR-associated protein 9 (CRISPR-Cas9) experiments have provided proof of concept for use in the functional investigation of mollusc gene sequences, showing for example that Pif (aragonite-binding) protein plays a significant role in structured nacre crystal growth and that theLsdia1gene sets shell chirality inLymnaea stagnalis. Much research has focused on the impacts of ocean acidification on molluscs. Initial studies were predominantly pessimistic for future molluscan biodiversity. However, more sophisticated experiments incorporating selective breeding and multiple generations are identifying subtle effects and that variability within mollusc genomes has potential for adaption to future conditions. Furthermore, we highlight recent historical studies based on museum collections that demonstrate a greater resilience of molluscs to climate change compared with experimental data. The future of mollusc research lies not solely with ecological investigations into biodiversity, and this review synthesizes knowledge across disciplines to understand biomineralization. It spans research ranging from evolution and development, through predictions of biodiversity prospects and future-proofing of aquaculture to identifying new biomimetic opportunities and societal benefits from recycling shell products.
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| 7. |
- De Wit, Pierre, 1978, et al.
(författare)
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Single nucleotide polymorphisms are suitable for assessing the success of restocking efforts of the European lobster (Homarus gammarus, L.)
- 2022
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Ingår i: Conservation Genetics Resources. - : Springer Science and Business Media LLC. - 1877-7252 .- 1877-7260. ; 14:1, s. 47-52
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Tidskriftsartikel (refereegranskat)abstract
- The European lobster (Homarus gammarus) forms the base of an important fishery along the coasts of Europe. However, stocks have been in decline for many years, prompting new regulations in the fishery and also restocking efforts. An important feature of any restocking effort is the assessment of success in the number of released juveniles that stay and become adult over time. Here, we tested the power of a single nucleotide polymorphism (SNP) DNA marker panel developed for population assignment to correctly infer parentage on the maternal side of lobster larvae, in the absence of known fathers, using lobsters included in a current restocking effort on the Swedish west coast. We also examined the power to reconstruct the unknown paternal genotypes, and examined the number of fathers for each larval clutch. We found that the 96-SNP panel, despite only containing 78 informative markers, allowed us to assign all larvae to the correct mother. Furthermore, with ten genotyped larvae or more, confident paternal genotypes could be reconstructed. We also found that 15 out of 17 clutches were full siblings, whereas two clutches had two fathers. To our knowledge, this is the first time a SNP panel of this size has been used to assess parentage in a crustacean restocking effort. Our conclusion is that the panel works well, and that it could be an important tool for the assessment of restocking success of H. gammarus in the future.
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| 8. |
- Doyle, Darragh, et al.
(författare)
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Transport and Barrier Functions in Rainbow Trout Trunk Skin Are Regulated by Environmental Salinity
- 2022
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Ingår i: Frontiers in Physiology. - : Frontiers Media SA. - 1664-042X. ; 13
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Tidskriftsartikel (refereegranskat)abstract
- The mechanisms underpinning ionic transport and barrier function have been relatively well characterised in amphibians and fish. In teleost fish, these processes have mostly been characterised in the gill and intestine. In contrast, these processes remain much less clear for the trunk skin of fish. In this study, we measured barrier function and active transport in the trunk skin of the rainbow trout, using the Ussing chamber technique. The effects of epithelial damage, skin region, salinity, and pharmacological inhibition were tested. Skin barrier function decreased significantly after the infliction of a superficial wound through the removal of scales. Wound healing was already underway after 3 h and, after 24 h, there was no significant difference in barrier function towards ions between the wounded and control skin. In relation to salinity, skin permeability decreased drastically following exposure to freshwater, and increased following exposure to seawater. Changes in epithelial permeability were accompanied by salinity-dependent changes in transepithelial potential and short-circuit current. The results of this study support the idea that barrier function in rainbow trout trunk skin is regulated by tight junctions that rapidly respond to changes in salinity. The changes in transepithelial permeability and short circuit current also suggest the presence of an active transport component. Immunostaining and selective inhibition suggest that one active transport component is an apical V-ATPase. However, further research is required to determine the exact role of this transporter in the context of the trunk skin.
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| 9. |
- Hedén, Ida, 1984, et al.
(författare)
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Dietary replacement of fishmeal with marine proteins recovered from shrimp and herring process waters promising in Atlantic salmon aquaculture
- 2023
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Ingår i: Aquaculture. - 0044-8486. ; 574
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Tidskriftsartikel (refereegranskat)abstract
- There is a general agreement that fish meal (FM) and fish oil (FO) are valuable resources for aquafeed, but that the production cannot keep the same pace as the current growth of the aquaculture industry. Therefore, there is a need to find alternative sources for lipids and protein. This study examines the possibility of using proteins recovered from seafood industry side stream waters as a complement to FM in feed for Atlantic salmon. To recover the proteins, herring and shrimp process side streams waters were flocculated then treated with dissolved air flotation (DAF), which is considered a gentle technology. Shrimp steam water was used to obtain shrimp protein (SP) and herring salt brine to obtain herring protein (HP). The recovered semi-solid protein fraction was spray dried and formulated into experimental diets at inclusion levels of 1.9–10% (dw/dw). Two feeding trials were conducted to investigate; 1) the properties of different flocculants to recover SP from shrimp steam water, alginate (Alg; 1.9% inclusion), carrageenan (Carr; 2.0% inclusion) and a synthetic flocculant from Kemira (Kem; 3.8% inclusion). 2) total or partial replacement of FM with SP-Alg (10% and no FM) and HP-Alg (3% and 8% FM). For both feeding trials each diet was provided to triplicate tanks (n = 31 and 30/tank) of Atlantic salmon with a start weight of 193 g (duration 10 weeks) and 304 g (duration 7 weeks) respectively. All fish showed similar feed intake, feed conversion ratio (on tank basis) and weight gain. The inclusion of SP or HP as total or partial replacement of FM did neither influence adiposity of the fish, as measured by condition factor, nor heposomatic index (HSI). No negative effect of the alternative protein could be found through histological examination of the intestine. In feeding trial two, diets did not affect the adaptive immune indicators CD8α and MHC II. The SP-Alg diet did not affect intestinal barrier and transporting functions, assessed using Ussing-chamber technology. However, HP-Alg affected the trans-epithelial resistance, which indicate that the intestinal barrier function could be affected by low inclusions. We conclude that from a biological perspective, SP recovered from shrimp steaming waters using Alg and DAF technology represents a new marine biomass with potential as a replacement for FM in Atlantic salmon feed. To diversify the possibility of using flocculants to retrieve proteins we also suggest further investigation of the potential to use Carr in larger inclusions.
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| 10. |
- Hedén, Ida, 1984, et al.
(författare)
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The role of environmental salinity on Na+-dependent intestinal amino acid uptake in rainbow trout (Oncorhynchus mykiss)
- 2022
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Ingår i: Scientific Reports. - : Springer Science and Business Media LLC. - 2045-2322. ; 12:1
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Tidskriftsartikel (refereegranskat)abstract
- Na+/K+-ATPases (NKA) in the basolateral membrane of the intestinal enterocytes create a Na+-gradient that drives both ion-coupled fluid uptake and nutrient transport. Being dependent on the same gradient as well as on the environmental salinity, these processes have the potential to affect each other. In salmonids, L-lysine absorption has been shown to be higher in freshwater (FW) than in seawater (SW) acclimated fish. Using electrophysiology (Ussing chamber technique), the aim was to explore if the decrease in L-lysine transport was due to allocation of the Na+-gradient towards ion-driven fluid uptake in SW, at the cost of amino acid transport. Intestinal NKA activity was higher in SW compared to FW fish. Exposure to ouabain, an inhibitor of NKA, decreased L-lysine transport. However, exposure to bumetanide and hydrochlorothiazide, inhibitors of Na+, K+, 2Cl−-co-transporter (NKCC) and Na+, Cl−-co-transporter (NCC) respectively, did not affect the rate of intestinal L-lysine transport. In conclusion, L-lysine transport is Na+-dependent in rainbow trout and the NKA activity and thus the available Na+-gradient increases after SW acclimation. This increased Na+-gradient is most likely directed towards osmoregulation, as amino acid transport is not compromised in SW acclimated fish.
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