Triplicate groups of triploid and diploid Atlantic salmon were fed diets with a low (LP, total P: 7.1 g kg⁻¹), medium (MP, total P: 9.4 g kg⁻¹) or high (HP, total P: 16.3 g kg⁻¹) phosphorous (P) ...level from first feeding (0.18 g) to transfer to sea water (~50 g, duration: 203 days) and subsequently fed a commercial diet in sea water for 426 days (~3 kg). This study examined the short‐ and long‐term effects of dietary P on freshwater performance (mortality, growth), vertebral deformities (radiology), bone cell activity (ALP and TRACP enzyme activity in vertebrae and scales, and fgf23, bgp and igf‐I relative gene expression in vertebrae), bone mineralization (ash content) and some parameters related to fish condition (heart and liver size). Irrespective of ploidy, at seawater transfer, fish fed the MP diet had significantly highest length and weight and those fed the LP diet significantly lowest length and weight, while those fed the HP diet had intermediate lengths and weights. Increased dietary phosphorus reduced deformities in both ploidies at seawater transfer; however, triploids fed the LP and MP diets had more deformities than diploids fed the respective diets, while there was no ploidy effect observed for fish fed the HP diet. The vertebral bone ash content at seawater transfer was significantly higher in diploids than in triploids when fed the MP diet only. Alkaline phosphatase (ALP) and tartrate‐resistant acid phosphatase (TRACP) enzyme activities and relative gene expression of bone hormones involved in metabolism of plasma phosphate (fgf23) and bone growth (bgp) were not affected by ploidy at seawater transfer, but by dietary P level; LP increased ALP activity and reduced TRACP activity and fgf23 and bgp expression levels in vertebral bone. In scales, LP increased both ALP and TRACP activity. At the termination of the seawater period, the group‐wise pattern in occurrence of vertebral deformities was the same as at seawater transfer. The present results on mortality, growth, bone mineralization and development of skeletal deformities all demonstrate that triploids have a higher P requirement than diploids in fresh water. This study shows that an optimalization of P nutrition for triploid Atlantic salmon can improve health and welfare and reduce down‐grading of triploid salmon.
Abstract
In the North Sea flatfish fishery, electric pulse trawls have been introduced to replace the conventional mechanical method. Pulse trawls reduce the fuel consumption, reduce adverse impact ...on the ecosystem but cause injuries in gadoids. We describe the design and electrical properties of pulse trawls currently in use and study the behavioural response and injuries in cod exposed to electrical pulses under controlled conditions. Pulse trawls operate at an average power of 0.7 kW m−1 beam length and a duty cycle of ∼2%. The electric field is heterogeneous with highest field strength occurring close to the conductors. Cod were exposed to three different pulse types for a range of field strengths, frequencies, and duty cycles. Two size classes were tested representing cod that escape through the meshes (11–17 cm) and market-sized cod that are retained in the net (34–56 cm). Cod exposed to a field strength of ≥37 V m−1 responded by moderate-to-strong muscular contractions. Some of the large cod (n = 260) developed haemorrhages and fractures in the spine, and haemal and neural arches in the tail part of the body. The probability of injuries increased with field strength and decreased when frequency was increased from 100 to 180 Hz. None of the small cod (n = 132) were injured and all survived. The field strength at the lateral boundaries of the trawl was too low to inflict injuries in cod.
This study sought to investigate whether a “natural diet” (mimicking the fatty acid composition of freshwater aquatic insects eaten by salmon parr) during the freshwater (FW) life stage of pre-smolt ...Atlantic salmon (Salmo salar L.) affected red blood cells and gill fatty acid composition as well as eicosanoid metabolism in gill during smolting at different temperatures. Before being transferred to seawater (SW), salmon parr were fed with a modified (MO) diet containing vegetable oils (rapeseed, palm, and linseed oils) supplemented with eicosapentaenoic acid (EPA) and arachidonic acid (ARA) to completely replace the fish oil (FO). Fatty acid composition in red blood cells and gill tissues was determined before SW transfer and six weeks after. Additionally, the expression of genes associated with eicosanoid metabolism and Na+/K+-ATPase (NKA) activity in salmon gill was examined at different temperatures before SW transfer and 24 h after. The results showed the changes in fatty acid composition, including sum monounsaturated fatty acids (MUFAs), docosahexaenoic acid (DHA), ARA, EPA, and sum n-6 polyunsaturated fatty acids (n-6 PUFA) in both red blood cells and gill tissues at the FW stage were consistent with the fatty acid profiles of the supplied MO and FO fish diets; however sum EPA and DHA composition exhibited opposite trends to those of the FO diet. The proportion of ARA, EPA, and n-6 PUFA increased, whereas sum MUFAs and DHA decreased in the red blood cells and gill tissues of MO-fed fish compared to those fed with the FO diet at FW stage. Additionally, 5-lipoxygenase-activating protein (Flap) expression was downregulated in MO-fed fish prior to SW transfer. During the process of SW transfer at different temperatures, the MO diet remarkably suppressed NKAα1a expression in MO-fed fish both at 12 and 16 °C. The MO diet also upregulated phospholipase A2 group IV (PLA2g4) expression in gills at 8, 12, and 16 °C, but suppressed phospholipase A2 group VI (PLA2g6) expression in gills at 12 °C compared to FO-fed fish at 12 °C and MO-fed fish at 8 °C. The MO diet also upregulated Cyclooxygenase 2 (Cox-2) expression at 8 °C compared to FO-fed fish and increased Arachidonate 5-lipoxygenase (5-Lox) expression in MO-fed fish at 16 °C compared to both FO-fed fish at 16 °C and MO-fed fish at 8 °C. Our study also determined that both SW transfer water temperatures and diets during the FW period jointly influenced the mRNA expression of PLA2g4, PLA2g6, and Lpl, whereas 5-Lox was more sensitive to dietary changes. In conclusion, the MO diet affected the fatty acid composition in gill and in red blood cells. When transferred to SW, dietary ARA supplementation could promote the bioavailability for eicosanoid synthesis in gill mainly via PLA2g4 activation, and potentially inhibit the stress and inflammatory response caused by different water temperatures through dietary EPA supplementation.
•Salmon parr were fed with a MO diet only containing vegetable oils supplemented with EPA and ARA.•The MO diet affected fatty acid composition synergistically in gills and red blood cells at FW stage.•A lasting impact of MO diet during FW stage on fatty acid composition after SW transfer.•The MO diet promoted the eicosanoid biosynthesis in gill during SW transfer.•The MO diet inhibited the inflammatory response during smoltification at different temperatures.
The aim of the current study was to investigate how freshwater diets impact on immunity in Atlantic salmon smolts in freshwater, during transfer to seawater and in post smolts during the seawater ...stage with and without pancreas disease (PD) infection. Three specific freshwater diets were prepared: (i) A diet similar in composition to commercial salmon freshwater diets (Standard diet); (ii) A diet composed of vegetable oils (rapeseed, palm and linseed oils) mimicking the fat composition in aquatic insects – the natural diet of wild salmon in freshwater (Fatty acid diet); (iii) A diet enriched with possible immune modulating amino acids including dl-methionine, l-lysine, l-threonine and taurine (Amino acid diet). After seawater transfer, all fish were fed the same commercial diet. Head kidneys were extracted, and their leukocytes isolated from smolts right before transfer to seawater, from post smolts one and six weeks after transfer to seawater, and from post smolts in seawater after 8 weeks of ongoing PD infection. In addition, to provoke bacterial or virus induced inflammation in vitro, the individual leukocyte suspension from all fish were stimulated by lipopolysaccharide (LPS) or polyinosinic acid: polycytidylic acid (PIC).
The transfer of smolts from fresh-to seawater changed the transcription of several types of genes. Particularly in isolates from fish fed the Standard or Fatty acid diet in freshwater, overall gene transcription (IL-1β, CD83, INF-γ, cox2, cd36, MGAT2, catalase) declined. However, the Amino acid diet stimulated the LPS induced gene transcription of IL-1β, CD83, Cox2, and INF-γ at this stage. In freshwater smolts, PIC stimulated leukocytes showed higher transcription level of Mx and viperin in the Fatty acid and Amino acid diet groups compared to the Standard diet group. In seawater post smolts, Mx and viperin responded similarly to PIC challenge in all diet groups. Furthermore, leukocytes isolated from PD infected fish, continued responding to PIC, regardless of freshwater diet.
•Modified Amino- and Fatty acid composition in freshwater diets can increase the immunological robustness of salmon parr towards virus infections during the freshwater phase.•The Amino acid composition in freshwater diets can be tailored to be protective against bacterial and oxidative damage in post smolts newly transferred to seawater.•When challenged with PIC, PD virus or PD virus + PIC, salmon immune cells use their energy to produce antiviral molecules, regardless of diet composition in freshwater.
The vitamin A (VA) concentration in salmon aquaculture feeds is varying and may lead to sublethal adverse effects. In this study, 135 g Atlantic salmon postsmolts were given eight diets in duplicates ...with 6, 12, 26, 55, 82, 112, 360 and 749 mg retinol (ROL) kg−1 for 116 days. Subsequently, fish given 6, 82 and 749 mg ROL kg−1 were transferred to a common net pen and given a standard commercial diet for further 28 weeks. Feed conversion rate, liver functionality and markers of VA homoeostasis were not negatively affected by dietary VA level, but chronic high VA intakes led to adverse effects on growth and bone health. In plasma, there was an antagonistic effect of dietary ROL on circulating 1,25 (OH)2 vitamin D3 (calcitriol). Moreover, a dose–response of VA on craniofacial deformities, condition factor and vertebral morphometry and mechanical strength was observed. Vertebral deformities were observed after 28 weeks on a standard diet and not immediately after the 116 days on the experimental diet. Elevated VA is a risk factor for bone deformities, and the dietary intake of VA should not exceed 37 mg ROL kg−1 body weight day−1 in Atlantic salmon postsmolts.
Genetic introgression of farmed salmon into wild populations can damage the genetic integrity of wild stocks and is therefore considered as an environmental threat. One possible solution is to induce ...sterility in farmed salmon. We have searched for proteins potentially essential for germline survival in Atlantic salmon. One of these is the argonaute protein Piwil1, known to be required for germ cell survival. To examine Piwil1 function in salmon, we induced indels in the N domain by CRISPR-Cas9. The encoded domain is present in all vertebrate Piwi proteins and has been linked to Tdrd1 protein interaction and PAZ lobe structure. The F0 founder generation of
piwil1
crispant males and females displayed a mosaic pattern of
piwil1
mutations, exhibiting highly mutated alleles (53%–97%) in their fin gDNA samples. In general,
piwil1
crispants carried germ cells, went through puberty and became fertile, although a transient and partial germ cell loss and delays during the spermatogenic process were observed in many male crispants, suggesting that Piwil1 functions during salmon spermatogenesis. By crossing highly mutated F0 founders, we produced F1 fish with a mixture of: loss-of-function alleles (
−
); functional in frame mutated alleles (
+
) and wt alleles (
+
). In F1, all
piwil1
−/−
fish lacked germ cells, while
piwil1
+/+
siblings showed normal ovaries and testes. Yet, most juvenile F1
piwil1
+/
−
males and females displayed an intermediate phenotype with a higher somatic/germ cell ratio without an increase in germ cell apoptosis, suggestive of a gene dose effect on the number of germ cells and/or insufficient replacement of lost germ cells in heterozygous fish. Interestingly, the two longest in-frame indels in the N domain also ensured germ cell loss. Hence, the loss of 4–6 aa in this region
Phe130
-
Ser136
may result in crucial changes of the protein structure, potentially affecting piRNA binding of the PAZ lobe, and/or affecting the binding of Piwil1 interacting proteins such as Tdrd protein, with critical consequences for the survival of primordial germ cells. In conclusion, we show that loss of
piwil1
leads to loss of germ cells in salmon and that part of the N domain of Piwil1 is crucial for its function.
Suboptimal egg incubation temperature is a risk factor for the development of skeletal deformities in teleosts. Triplicate diploid and triploid Atlantic salmon, Salmo salar L., egg batches were ...incubated at 6, 8 and 10 °C up until first feeding, whereupon fish were reared on a natural temperature before examination for externally visible skeletal deformities (jaw and spine) and radiographed for vertebral deformities and morphology at the parr stage. Increasing incubation temperatures and triploidy increased the number of fish showing one or more deformed vertebrae. Triploids had significantly higher mean vertebrae cranio‐caudal length (L) and dorsal‐ventral height (H) ratio at 6 and 10 °C than diploids, but triploidy had no effect on mean vertebrae centra area. Triploids demonstrated an increase in lower jaw deformities with increased incubation temperature, whereas jaw deformities were rare in diploids. Fish incubated at 10 °C had a significantly lower mean vertebral number than fish incubated at 6 °C, and triploids had lower mean vertebral numbers than diploids. Diploid fish with 58 vertebrae had a significantly higher mean vertebral centra area than fish with 59 vertebrae, but vertebral number did not affect the mean vertebral L/H ratio. The results are discussed with respect to the welfare and production of farmed salmonids.
This study investigates the effects of water temperature (T) on vaccine-induced abdominal lesions (i.p. injection with oil-adjuvant vaccine) and vertebral deformities in Atlantic salmon. Quadruple ...groups of vaccinated (V) or unvaccinated (U) underyearling smolts were reared in tanks under four different temperature regimes for 6 weeks in fresh water (FW) followed by 6 weeks in sea water (SW). The four different T regimes were 10 °C FW - 10 °C SW (10-10), 10 °C FW - 16 °C SW (10-16), 16 °C FW - 10 °C SW (16-10) and 16 °C FW - 16 °C SW (16-16). After the temperature regimes were finished, the fish were group-tagged and transferred to a common sea cage for on-growth until harvest size. At termination, weight was significantly affected by both T and V, while lesion score and deformities were affected by T only. The weight difference between the largest and smallest U group was 20.3% (16-10 U: 2.4 kg, 10-16 U: 1.89 kg), while the largest difference between U and V fish within a T regime was 28.7% (16-16 U: 2.1 kg, 16-16 V: 1.5 kg). Fish from the 16-16, 16-10 and 10-16 regimes had a significant higher lesion score than those from the 10-10 regime. Fish from the 10-16 and 16-16 regimes displayed a significantly higher prevalence of vertebral deformities (palpation : 13-27%, radiology: 88-94%) than fish from the 10-10 and 16-10 regimes (palpation: 2-3%, radiology: 27-65%). Vertebra number 26 (located beneath the dorsal fin) was the most frequently affected vertebra in smolts, while vertebra number 43 (located above the anal fin) was most frequently affected in adults.
The use of sterile triploid stock in the Atlantic salmon (
Salmo salar, L) farming industry is the only commercially available means to prevent the ecological impact of domesticated escapees. This ...study compared the seawater (SW) performance and deformity prevalence of diploid and triploid post-smolts from 2 full-sib families produced out-of-season. Triploids completed smoltification 4
weeks earlier and at a significantly higher body-weight. Growth and survival in SW were not significantly affected by ploidy. The incidence of external deformities, dominated by jaw malformation, was ~
12% in triploids and below 5% in diploids. Vertebral deformities were more prevalent in the fastest growing triploid family only. Heart morphometry differed between ploidies which may relate to a higher cardiac workload in triploids. No clear alteration of the gill apparatus was detected. The most significant detrimental effect of triploidy was on the rate and severity of cataract that were observed from August onward (50% and 92% of diploids and triploids respectively affected after 1-year in SW). At that time, cataracts were diagnosed by histological examinations as irreversible with a probable osmotic origin which could arise from factors such as water quality, nutritional deficiencies or thermal variations. This study warrants further research aiming at adapting rearing practices to the needs of triploid stocks as to improve their performance and welfare.
An earlier study demonstrated that under-yearling (0+) Atlantic salmon (Salmo salar L.) smolt had a lower vertebral mineral content and mechanical strength and higher prevalence of vertebral ...deformities than 1+ smolt during the early seawater (SW) phase. The present study aimed to examine if commercial extruded high-energy diets need to be supplemented additional minerals for proper bone mineralization and prevention of bone deformities in fast growing 0 + smolts. We studied vertebral morphology with radiology, and bone mineral content and mechanical strength in 60 g 0+ smolt fed diets with a normal (NM) or elevated (HM) bone mineral (P and Ca) contents from SW transfer (week 0) until 10 times weight increase at week 17. Thereafter, both groups were fed a commercial diet until a mean slaughter weight of 4100 g after 57 week. There were no differences in body weight and length between the dietary groups during the study, while the condition factor differed significantly at the final sampling (NM 1.40; HM 1.29). The most common bone deformity observed was compressions in the tail region of the vertebral column. Lower incidences of vertebral deformities (percent individuals with one or more deformed vertebrae) was observed in the HM group in week 17 (HM 20%; NM 47%) and week 57 (HM 37%; NM 73%), also reflected by higher vertebral length/dorso-ventral diameter ratio in weeks 17 (HM 0.99; MN 0.92) and 57 (HM 0.97; NM 0.88). The HM group had significantly higher vertebral mineral content (HM 550 g kg⁻¹; NM 480 g kg⁻¹) and mechanical strength (HM 9050 g mm⁻¹; NM 4600 g mm⁻¹) than the NM group after 8 week feeding. Plasma levels of Ca, P and D-vitamin metabolites recorded in week 8 reflected changes in P homeostasis, but could not explain the preventive effect of the HM diet on development of bone deformities. The results suggest that elevated dietary mineral content during the early SW phase may reduce the prevalence of vertebral deformities in fast growing 0 + salmon smolts.
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