Arctic charr (Salvelinus alpinus) were fed for 99 days on experimental diets with 40% of fish mea... more Arctic charr (Salvelinus alpinus) were fed for 99 days on experimental diets with 40% of fish meal replaced, on a crude protein basis, with intact yeast (Saccharomyces cerevisiae) (ISC), extracted yeast (ESC), Rhizopus oryzae fungus (RHO) or de-shelled blue mussels (Mytilus edulis) (MYE). The fish were evaluated for growth performance, nutrient digestibility and fish intestinal function. Growth performance, retention of crude protein and sum of amino acids were not affected in fish fed diets ISC or MYE compared with those fed the reference (REF) diet. However, fish fed diet ISC displayed decreased digestibility of crude protein and indispensable amino acids and decreased intestinal barrier function compared with fish fed the REF diet. Fish fed diet ESC exhibited decreased growth performance and protein retention, but had comparable digestibility to fish fed the REF diet. Fish fed diets MYE and RHO showed similar performance in terms of growth, nutrient digestibility and intestinal b...
Aquaculture is a growing industry, increasing the need for understanding host-pathogen interactio... more Aquaculture is a growing industry, increasing the need for understanding host-pathogen interactions in fish. The skin and mucosal surfaces, covered by a mucus layer composed of mucins, is the first point of contact between fish and pathogens. Highly O-glycosylated mucins have been shown to be an important part of the defense against pathogens, and pathogens bind to host surfaces using lectin-like adhesins. However, knowledge of piscine O-glycosylation is very limited. We characterized mucin O-glycosylation of five freshwater acclimated Atlantic salmon, using mass spectrometry. Of the 109 O-glycans found, most were sialylated and differed in distribution among skin, pyloric ceca, and proximal and distal intestine. Skin O-glycans were shorter (2-6 residues) and less diverse (33 structures) than intestinal O-glycans (2-13 residues, 93 structures). Skin mucins carried O-glycan cores 1, 2, 3, and 5 and three types of sialic acids (Neu5Ac, Neu5Gc, and Kdn) and had sialyl-Tn as the predominant structure. Intestinal mucins carried only cores 1, 2, and 5, Neu5Ac was the only sialic acid present, and sialylated core 5 was the most dominant structure. This structural characterization can be used for identifying structures of putative importance in host-pathogen interactions for further testing in biological assays and disease intervention therapies.
ABSTRACT The intestinal epithelium of fish is a selective barrier faced with a complex task: bloc... more ABSTRACT The intestinal epithelium of fish is a selective barrier faced with a complex task: blocking pathogens and harmful substances while simultaneously allowing nutrient and water absorption. Therefore, an appropriate function and regulation of the intestinal barrier is essential for maintaining the intestinal health and the health of the whole animal. Intestinal health is maintained with cooperation between the different intestinal barriers (i.e., the extrinsic mucus barrier, the intrinsic physical epithelial barrier, and the immunological barrier). This chapter focuses on describing the constituents of the physical epithelial barrier, how to assess intestinal permeability and selectivity and to give examples on how intestinal permeability and selectivity is affected and regulated by factors in the external and internal environment, and how this is related to intestinal health.
This study investigated the expression of ion transporters involved in intestinal fluid absorptio... more This study investigated the expression of ion transporters involved in intestinal fluid absorption and presents evidence for developmental changes in abundance and tissue distribution of these transporters during smoltification and seawater (SW) acclimation of Atlantic salmon Salmo salar. Emphasis was placed on Na(+) , K(+) -ATPase (NKA) and Na(+) , K(+) , Cl(-) co-transporter (NKCC) isoforms, at both transcriptional and protein levels, together with transcription of chloride channel genes. The nka α1c was the dominant isoform at the transcript level in both proximal and distal intestines; also, it was the most abundant isoform expressed in the basolateral membrane of enterocytes in the proximal intestine. This isoform was also abundantly expressed in the distal intestine in the lower part of the mucosal folds. The protein expression of intestinal Nkaα1c increased during smoltification. Immunostaining was localized to the basal membrane of the enterocytes in freshwater (FW) fish, an...
Aeromonas salmonicida subsp. salmonicida infection, also known as furunculosis disease, is associ... more Aeromonas salmonicida subsp. salmonicida infection, also known as furunculosis disease, is associated with high morbidity and mortality in salmonid aquaculture. The first line of defense the pathogen encounters is the mucus layer, which is predominantly comprised of secreted mucins. Here we isolated and characterized mucins from the skin and intestinal tract of healthy Atlantic salmon and studied how A. salmonicida bound to them. The mucins from the skin, pyloric ceca, and proximal and distal intestine mainly consisted of mucins soluble in chaotropic agents. The mucin density and mucin glycan chain length from the skin were lower than were seen with mucin from the intestinal tract. A. salmonicida bound to the mucins isolated from the intestinal tract to a greater extent than to the skin mucins. The mucins from the intestinal regions had higher levels of sialylation than the skin mucins. Desialylating intestinal mucins decreased A. salmonicida binding, whereas desialylation of skin mucins resulted in complete loss of binding. In line with this, A. salmonicida also bound better to mammalian mucins with high levels of sialylation, and N-acetylneuraminic acid appeared to be the sialic acid whose presence was imperative for binding. Thus, sialylated structures are important for A. salmonicida binding, suggesting a pivotal role for sialylation in mucosal defense. The marked differences in sialylation as well as A. salmonicida binding between the skin and intestinal tract suggest interorgan differences in the host-pathogen interaction and in the mucin defense against A. salmonicida.
In high intensive fish production systems, hyperoxygenation and reduced flow are often used to sa... more In high intensive fish production systems, hyperoxygenation and reduced flow are often used to save water and increase the holding capacity. This commonly used husbandry practice has been shown to be stressful to fish and increase mortality after infectious pancreatic necrosis virus (IPNV) challenge, but the cause and effect relationship is not known. Salmonids are particularly sensitive to stress during smoltification and the first weeks after seawater (SW) transfer. This work aimed at investigating the impact of hyperoxygenation combined with reduced flow in fresh water (FW), on the intestinal barrier in FW as well as during later life stages in SW. It further aims at investigating the role of the intestinal barrier during IPNV challenge and possible secondary infections. Hyperoxygenation in FW acted as a stressor as shown by significantly elevated plasma cortisol levels. This stressful husbandry condition tended to increase paracellular permeability (P(app)) as well as translocation of Aeromonas salmonicida in the posterior intestine of Atlantic salmon. After transfer to SW and subsequent IPNV challenge, intestinal permeability, as shown by P(app), and translocation rate of A. salmonicida increased in the anterior intestine, concomitant with further elevation in plasma cortisol levels. In the anterior intestine, four of five fish displayed alterations in intestinal appearance. In two of five fish, IPNV caused massive necrosis with significant loss of cell material and in a further two fish, IPNV caused increased infiltration of lymphocytes into the epithelium and granulocytes in the lamina propria. Hyperoxygenation and reduced flow in the FW stage may serve as stressors with impact mainly during later stages of development. Fish with an early history of hyperoxygenation showed a higher stress response concomitant with a disturbed intestinal barrier function, which may be a cause for the increased susceptibility to IPNV infection and increased susceptibility to secondary infections.
Previous studies have shown that specific binding of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) and 2... more Previous studies have shown that specific binding of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) and 24,25-dihydroxyvitamin D3 (24,25(OH)2D3) to enterocyte basolateral membranes (BLM), as well as circulating concentrations, is affected in response to changes in environmental salinity. It is not known if the production of 1,25(OH)2D3 and 24,25(OH)2D3 is affected by environmental salinity. The aim of the present study was to measure the in vitro production of [3H]-1,25(OH)2D3 and [3H]-24,25(OH)2D3 in fresh water (FW) and after 1, 2, 3, and 7 days after transfer to seawater (SW). Pooled sub-cellular fractions (mitochondria and microsomes) from liver or kidney was incubated with [3H]-25(OH)D3 and the produced metabolites were separated using HPLC. Hepatic production of [3H]-1,25(OH)2D3 was decreased after 24h in SW. This was followed by an up-regulation after 48h and a second, slower decrease in production rate which leveled out after 7 days in SW. The production rate in SW was lower than the original rate in FW-adapted fish. For hepatic [3H]-24,25(OH)2D3 production the pattern was reversed. Renal production of [3H]-24,25(OH)2D3 increased significantly during the period of SW acclimation. These results suggest that environmental salinity regulates the production rate of the two antagonizing calcium regulatory hormones; 1,25(OH)2D3 and 24,25(OH)2D3. This gives further evidence to the hypothesis that there is a physiological regulation and a differentiated importance of 1,25(OH)2D3 and 24,25(OH)2D3 in relation to environmental calcium concentrations.
Arctic charr (Salvelinus alpinus) were fed for 99 days on experimental diets with 40% of fish mea... more Arctic charr (Salvelinus alpinus) were fed for 99 days on experimental diets with 40% of fish meal replaced, on a crude protein basis, with intact yeast (Saccharomyces cerevisiae) (ISC), extracted yeast (ESC), Rhizopus oryzae fungus (RHO) or de-shelled blue mussels (Mytilus edulis) (MYE). The fish were evaluated for growth performance, nutrient digestibility and fish intestinal function. Growth performance, retention of crude protein and sum of amino acids were not affected in fish fed diets ISC or MYE compared with those fed the reference (REF) diet. However, fish fed diet ISC displayed decreased digestibility of crude protein and indispensable amino acids and decreased intestinal barrier function compared with fish fed the REF diet. Fish fed diet ESC exhibited decreased growth performance and protein retention, but had comparable digestibility to fish fed the REF diet. Fish fed diets MYE and RHO showed similar performance in terms of growth, nutrient digestibility and intestinal b...
Aquaculture is a growing industry, increasing the need for understanding host-pathogen interactio... more Aquaculture is a growing industry, increasing the need for understanding host-pathogen interactions in fish. The skin and mucosal surfaces, covered by a mucus layer composed of mucins, is the first point of contact between fish and pathogens. Highly O-glycosylated mucins have been shown to be an important part of the defense against pathogens, and pathogens bind to host surfaces using lectin-like adhesins. However, knowledge of piscine O-glycosylation is very limited. We characterized mucin O-glycosylation of five freshwater acclimated Atlantic salmon, using mass spectrometry. Of the 109 O-glycans found, most were sialylated and differed in distribution among skin, pyloric ceca, and proximal and distal intestine. Skin O-glycans were shorter (2-6 residues) and less diverse (33 structures) than intestinal O-glycans (2-13 residues, 93 structures). Skin mucins carried O-glycan cores 1, 2, 3, and 5 and three types of sialic acids (Neu5Ac, Neu5Gc, and Kdn) and had sialyl-Tn as the predominant structure. Intestinal mucins carried only cores 1, 2, and 5, Neu5Ac was the only sialic acid present, and sialylated core 5 was the most dominant structure. This structural characterization can be used for identifying structures of putative importance in host-pathogen interactions for further testing in biological assays and disease intervention therapies.
ABSTRACT The intestinal epithelium of fish is a selective barrier faced with a complex task: bloc... more ABSTRACT The intestinal epithelium of fish is a selective barrier faced with a complex task: blocking pathogens and harmful substances while simultaneously allowing nutrient and water absorption. Therefore, an appropriate function and regulation of the intestinal barrier is essential for maintaining the intestinal health and the health of the whole animal. Intestinal health is maintained with cooperation between the different intestinal barriers (i.e., the extrinsic mucus barrier, the intrinsic physical epithelial barrier, and the immunological barrier). This chapter focuses on describing the constituents of the physical epithelial barrier, how to assess intestinal permeability and selectivity and to give examples on how intestinal permeability and selectivity is affected and regulated by factors in the external and internal environment, and how this is related to intestinal health.
This study investigated the expression of ion transporters involved in intestinal fluid absorptio... more This study investigated the expression of ion transporters involved in intestinal fluid absorption and presents evidence for developmental changes in abundance and tissue distribution of these transporters during smoltification and seawater (SW) acclimation of Atlantic salmon Salmo salar. Emphasis was placed on Na(+) , K(+) -ATPase (NKA) and Na(+) , K(+) , Cl(-) co-transporter (NKCC) isoforms, at both transcriptional and protein levels, together with transcription of chloride channel genes. The nka α1c was the dominant isoform at the transcript level in both proximal and distal intestines; also, it was the most abundant isoform expressed in the basolateral membrane of enterocytes in the proximal intestine. This isoform was also abundantly expressed in the distal intestine in the lower part of the mucosal folds. The protein expression of intestinal Nkaα1c increased during smoltification. Immunostaining was localized to the basal membrane of the enterocytes in freshwater (FW) fish, an...
Aeromonas salmonicida subsp. salmonicida infection, also known as furunculosis disease, is associ... more Aeromonas salmonicida subsp. salmonicida infection, also known as furunculosis disease, is associated with high morbidity and mortality in salmonid aquaculture. The first line of defense the pathogen encounters is the mucus layer, which is predominantly comprised of secreted mucins. Here we isolated and characterized mucins from the skin and intestinal tract of healthy Atlantic salmon and studied how A. salmonicida bound to them. The mucins from the skin, pyloric ceca, and proximal and distal intestine mainly consisted of mucins soluble in chaotropic agents. The mucin density and mucin glycan chain length from the skin were lower than were seen with mucin from the intestinal tract. A. salmonicida bound to the mucins isolated from the intestinal tract to a greater extent than to the skin mucins. The mucins from the intestinal regions had higher levels of sialylation than the skin mucins. Desialylating intestinal mucins decreased A. salmonicida binding, whereas desialylation of skin mucins resulted in complete loss of binding. In line with this, A. salmonicida also bound better to mammalian mucins with high levels of sialylation, and N-acetylneuraminic acid appeared to be the sialic acid whose presence was imperative for binding. Thus, sialylated structures are important for A. salmonicida binding, suggesting a pivotal role for sialylation in mucosal defense. The marked differences in sialylation as well as A. salmonicida binding between the skin and intestinal tract suggest interorgan differences in the host-pathogen interaction and in the mucin defense against A. salmonicida.
In high intensive fish production systems, hyperoxygenation and reduced flow are often used to sa... more In high intensive fish production systems, hyperoxygenation and reduced flow are often used to save water and increase the holding capacity. This commonly used husbandry practice has been shown to be stressful to fish and increase mortality after infectious pancreatic necrosis virus (IPNV) challenge, but the cause and effect relationship is not known. Salmonids are particularly sensitive to stress during smoltification and the first weeks after seawater (SW) transfer. This work aimed at investigating the impact of hyperoxygenation combined with reduced flow in fresh water (FW), on the intestinal barrier in FW as well as during later life stages in SW. It further aims at investigating the role of the intestinal barrier during IPNV challenge and possible secondary infections. Hyperoxygenation in FW acted as a stressor as shown by significantly elevated plasma cortisol levels. This stressful husbandry condition tended to increase paracellular permeability (P(app)) as well as translocation of Aeromonas salmonicida in the posterior intestine of Atlantic salmon. After transfer to SW and subsequent IPNV challenge, intestinal permeability, as shown by P(app), and translocation rate of A. salmonicida increased in the anterior intestine, concomitant with further elevation in plasma cortisol levels. In the anterior intestine, four of five fish displayed alterations in intestinal appearance. In two of five fish, IPNV caused massive necrosis with significant loss of cell material and in a further two fish, IPNV caused increased infiltration of lymphocytes into the epithelium and granulocytes in the lamina propria. Hyperoxygenation and reduced flow in the FW stage may serve as stressors with impact mainly during later stages of development. Fish with an early history of hyperoxygenation showed a higher stress response concomitant with a disturbed intestinal barrier function, which may be a cause for the increased susceptibility to IPNV infection and increased susceptibility to secondary infections.
Previous studies have shown that specific binding of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) and 2... more Previous studies have shown that specific binding of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) and 24,25-dihydroxyvitamin D3 (24,25(OH)2D3) to enterocyte basolateral membranes (BLM), as well as circulating concentrations, is affected in response to changes in environmental salinity. It is not known if the production of 1,25(OH)2D3 and 24,25(OH)2D3 is affected by environmental salinity. The aim of the present study was to measure the in vitro production of [3H]-1,25(OH)2D3 and [3H]-24,25(OH)2D3 in fresh water (FW) and after 1, 2, 3, and 7 days after transfer to seawater (SW). Pooled sub-cellular fractions (mitochondria and microsomes) from liver or kidney was incubated with [3H]-25(OH)D3 and the produced metabolites were separated using HPLC. Hepatic production of [3H]-1,25(OH)2D3 was decreased after 24h in SW. This was followed by an up-regulation after 48h and a second, slower decrease in production rate which leveled out after 7 days in SW. The production rate in SW was lower than the original rate in FW-adapted fish. For hepatic [3H]-24,25(OH)2D3 production the pattern was reversed. Renal production of [3H]-24,25(OH)2D3 increased significantly during the period of SW acclimation. These results suggest that environmental salinity regulates the production rate of the two antagonizing calcium regulatory hormones; 1,25(OH)2D3 and 24,25(OH)2D3. This gives further evidence to the hypothesis that there is a physiological regulation and a differentiated importance of 1,25(OH)2D3 and 24,25(OH)2D3 in relation to environmental calcium concentrations.
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