Deformación de la aleta caudal en alevín de salmón del atlántico (Salmo salar), en etapa de saco vitelino / Deformation of the caudal fin in atlantic salmon fry (Salmo salar), in the yolk sac stage

RESUMEN: En este estudio se analiza una deformación que afectó la aleta caudal de los alevines de salmón del Atlántico (Salmo salar) y que les proporcionó un aspecto de "cola aguzada". Al momento de la eclosión se observaron completamente normales pero la deformación se detectó con posterioridad. El objetivo de este trabajo es describir los cambios anatómicos e histológicos de la aleta caudal deformada del alevín de salmón del Atlántico (Salmo salar). Para esto se utilizaron 60 alevines para realizar análisis de laboratorio y descartar la presencia de patógenos virales o bacterianos. Otros 60 alevines con un estado de desarrollo entre 600 y 700 Unidades térmicas acumuladas (UTAs) fueron anestesiados con Benzocaína 5 %, fijados en formalina al 10 % pesados y medidos. De estos un grupo de 30 alevines (15 normales y 15 deformes) fueron sometidos a la técnica de Hanken y Wassersug para evaluar Lepidotriquias. Los otros 30 alevines (15 normales y 15 deformes), fueron procesados mediante las técnicas histoquímicas: H&E/azul de Alcián para evaluar las características histológicas generales. Adicionalmente se utilizó técnicas inmunohistoquímicas para reconocer la ubicación y la presencia de los centros de señales Sonic hedgehog (Shh) para la formación de Lepidotriquias. A los valores obtenidos para las variables cuantitativas peso y longitud de cuerpo, largo y ancho de aleta caudal, se les realizó estadística descriptiva y fueron sometidos a prueba de normalidad de Shapiro-Wilk. Las diferencias observadas entre peces normales y deformes, fueron analizadas mediante prueba t de Student o U de Mann Whitney, utilizando el paquete estadístico IBM SPSS 20.0. La deformación se observó desde las 600 UTA. El peso de los alevines deformes fue similar al de los peces normales (p>0,05), lo mismo sucedió con la longitud de la aleta (p>0,05). Por el contrario, el ancho de la aleta de los deformes fue muy reducida (p<0,05). El fenotipo de aleta aguzada presentó un cambio en la integridad de los bordes, fracturas de Lepidotriquias. La epidermis de la aleta caudal de los alevines deformados presentó sus centros de señalización Shh activos, pero el blastema interrayos presentó vasodilatación, congestión y hemorragias. La presentación de este caso se relacionó con incrementos bruscos de temperatura peri eclosional.

SUMMARY: This study analyzes the deformity of Atlantic salmon fry (Salmo salar) caudal fin, which gives it a "pointed tail" appearance. Although at hatching specimens were normal, the deformities were detected later. The objective of this work is to describe the anatomical and histological changes of the deformed caudal fin of the Atlantic salmon fry (Salmo salar). In this analysis we used 60 specimens for laboratory analyses, to rule out the presence of viral or bacterial pathogens. Another 60 fry, developmental stage between 600 and 700 Accumulated Thermal Units (UTAs) were anesthetized with 5 % Benzocaine, fixed in 10 % formalin, weighed and measured. Of these, a group of 30 fry (15 normal and 15 deformed) underwent the Hanken and Wassersug technique to evaluate lepidotrychia or dermal rays. The remaining 30 fry (15 normal and 15 deformed) were processed using H & E / Alcián blue histochemical techniques to evaluate general histological characteristics. Additionally, immunohistochemical techniques were used to determine the location and presence of Sonic hedgehog (Shh) signal centers for lepidotrychia development. The values obtained for the quantitative variables body weight, length and width of the tail fin were described using the arithmetic mean and standard deviation. The deformity was observed from 600 UTA. Weight of deformed fry was less than normal fish, length of the fin was similar in normal and deformed fish. In contrast, width of the deformed fin was significantly reduced. The sharp fin phenotype presented a change in the conformity of the edges, lepidotrychia fractures. The epidermis presented active Shh signaling centers, but the interray blastema showed vasodilation, congestion and hemorrhages. The presentation of this case was related to sudden increases in perieclosional temperature.

Vertebral column deformity with curved cross-stitch vertebrae in Norwegian seawater-farmed Atlantic salmon, Salmo salar L.

Pathological changes in the vertebral column of farmed Atlantic salmon in Norway have been reported since the 1990s. Based on the characteristic radiographic findings, we here present a vertebral column deformity named "curved cross-stitch vertebrae" that mainly affects the middle aspect of the vertebral column. Sixty fish, from the west/northwest coast of mid-Norway, were sampled at slaughter and examined by radiography, computed tomography (CT), necropsy, macrophotography, and histology. The vertebral deformities were radiographically graded as mild, moderate, or marked. The main differences between these grades of changes were defined by increased curving of the peripheries of endplates, reduced intervertebral spaces, and vertical displacement of the vertebrae. The curved rims of endplates were located peripheral to a continuous and approximately circular borderline. The CT studies revealed small, multifocal, hypo-attenuating, round to crescent-shaped areas in the notochord, compatible with the presence of gas. Additionally, histology revealed that the axial parts of endplates had circular zones with perforations, through which either notochordal tissue prolapsed into the vertebrae or vascularized fibrochondroid proliferations extended from the vertebrae into the notochord. Inflammation was present in many vertebral bodies. To the best of our knowledge, this is the first report of gas in the notochord of fish.

Enhanced micronutrient supplementation in low marine diets reduced vertebral malformation in diploid and triploid Atlantic salmon (Salmo salar) parr, and increased vertebral expression of bone biomarker genes in diploids.

Previously we showed that, for optimum growth, micronutrient levels should be supplemented above current National Research Council (2011) recommendations for Atlantic salmon when they are fed diets formulated with low levels of marine ingredients. In the present study, the impact of graded levels (100, 200, 400%) of a micronutrient package (NP) on vertebral deformities and bone gene expression were determined in diploid and triploid salmon parr fed low marine diets. The prevalence of radiologically detectable spinal deformities decreased with increasing micronutrient supplementation in both ploidy. On average, triploids had a higher incidence of spinal deformity than diploids within a given diet. Micronutrient supplementation particularly reduced prevalence of fusion deformities in diploids and compression and reduced spacing deformities in triploids. Prevalence of affected vertebrae within each spinal region (cranial, caudal, tail and tail fin) varied significantly between diet and ploidy, and there was interaction. Prevalence of deformities was greatest in the caudal region of triploids and the impact of graded micronutrient supplementation in reducing deformities also greatest in triploids. Diet affected vertebral morphology with length:height (L:H) ratio generally increasing with level of micronutrient supplementation in both ploidy with no difference between ploidy. Increased dietary micronutrients level in diploid salmon increased the vertebral expression of several bone biomarker genes including bone morphogenetic protein 2 (bmp2), osteocalcin (ostcn), alkaline phosphatase (alp), matrix metallopeptidase 13 (mmp13), osteopontin (opn) and insulin-like growth factor 1 receptor (igf1r). In contrast, although some genes showed similar trends in triploids, vertebral gene expression was not significantly affected by dietary micronutrients level. The study confirmed earlier indications that dietary micronutrient levels should be increased in salmon fed diets with low marine ingredients and that there are differences in nutritional requirements between ploidies.

Growth and development of skeletal anomalies in diploid and triploid Atlantic salmon (Salmo salar) fed phosphorus-rich diets with fish meal and hydrolyzed fish protein.

Diploid and triploid Atlantic salmon, Salmo salar were fed high-protein, phosphorus-rich diets (56-60% protein; ca 18g phosphorus kg-1 diet) whilst being reared at low temperature from start-feeding until parr-smolt transformation. Performances of salmon fed diets based on fish meal (STD) or a mix of fishmeal and hydrolysed fish proteins (HFM) as the major protein sources were compared in terms of mortality, diet digestibility, growth and skeletal deformities. Separate groups of diploids and triploids were reared in triplicate tanks (initially 3000 fish per tank; tank biomass ca. 620 g) from 0-2745 degree-days post-start feeding (ddPSF). Growth metrics (weight, length, condition factor) were recorded at ca. 4 week intervals, external signs of deformities to the operculum, jaws and spinal column were examined in parr sampled at 1390 ddPSF, and external signs of deformity and vertebral anomalies (by radiography) were examined in fish sampled at the end of the trial (2745 ddPSF). The triploid salmon generally had a lower mass per unit length, i.e. lower condition factor, throughout the trial, but this did not seem to reflect any consistent dietary or ploidy effects on either dietary digestibility or the growth of the fish. By the end of the trial fish in all treatment groups had achieved a weight of 50+ g, and had completed the parr-smolt transformation. The triploids had slightly, but significantly, fewer vertebrae (Triploids STD 58.74 ± 0.10; HFM 58.68 ± 0.05) than the diploids (Diploids STD 58.97 ± 0.14; HFM 58.89 ± 0.01), and the incidence of skeletal (vertebral) abnormalities was higher in triploids (Triploids STD 31 ± 0.90%; HFM 15 ± 1.44%) than in diploids (Diploids STD 4 ± 0.80%; HFM 4 ± 0.83%). The HFM diet gave a significant reduction in the numbers of triploid salmon with vertebral anomalies in comparison with the triploids fed the STD diet possibly as a result of differences in phosphorus bioavailability between the two diets. Overall, the incidence of skeletal deformities was lower than reported in previous studies (Diploids 20+%, Triploids 40+%), possibly as a result of the combination of rearing at low-temperature and phosphorus-rich diets being used in the present study.

Rapid growth causes abnormal vaterite formation in farmed fish otoliths.

Sagittal otoliths are essential components of the sensory organs that enable all teleost fish to hear and maintain balance, and are primarily composed of calcium carbonate. A deformity, where aragonite (the normal crystal form) is replaced with vaterite, was first noted over 50 years ago but its underlying cause is unresolved. We evaluated the prevalence of vateritic otoliths from two captive rearing studies which suggested that fast growth, due to environmental rather than genetic control, led to vaterite development. We then tested this by varying light and temperature to create phenotypes with different growth rates, which resulted in fast growers (5 times larger) having 3 times more vaterite than slow growers. A decrease in either the ratio of otolith matrix proteins (otolin-1/OMM-64) or [Ca2+]/[CO32-] may explain why fast growth causes vaterite deposition. As vaterite decreases hearing sensitivity, reducing growth rates in hatcheries may improve the welfare of farmed fish and increase the success of conservation efforts.

Multigenic Delineation of Lower Jaw Deformity in Triploid Atlantic Salmon (Salmo salar L.).

Lower jaw deformity (LJD) is a skeletal anomaly affecting farmed triploid Atlantic salmon (Salmo salar L.) which leads to considerable economic losses for industry and has animal welfare implications. The present study employed transcriptome analysis in parallel with real-time qPCR techniques to characterise for the first time the LJD condition in triploid Atlantic salmon juveniles using two independent sample sets: experimentally-sourced salmon (60 g) and commercially produced salmon (100 g). A total of eleven genes, some detected/identified through the transcriptome analysis (fbn2, gal and gphb5) and others previously determined to be related to skeletal physiology (alp, bmp4, col1a1, col2a1, fgf23, igf1, mmp13, ocn), were tested in the two independent sample sets. Gphb5, a recently discovered hormone, was significantly (P < 0.05) down-regulated in LJD affected fish in both sample sets, suggesting a possible hormonal involvement. In-situ hybridization detected gphb5 expression in oral epithelium, teeth and skin of the lower jaw. Col2a1 showed the same consistent significant (P < 0.05) down-regulation in LJD suggesting a possible cartilaginous impairment as a distinctive feature of the condition. Significant (P < 0.05) differential expression of other genes found in either one or the other sample set highlighted the possible effect of stage of development or condition progression on transcription and showed that anomalous bone development, likely driven by cartilage impairment, is more evident at larger fish sizes. The present study improved our understanding of LJD suggesting that a cartilage impairment likely underlies the condition and col2a1 may be a marker. In addition, the involvement of gphb5 urges further investigation of a hormonal role in LJD and skeletal physiology in general.

Concurrence of lower jaw skeletal anomalies in triploid Atlantic salmon (Salmo salar L.) and the effect on growth in freshwater.

Triploid Atlantic salmon populations are associated with higher prevalence of lower jaw skeletal anomalies affecting fish performance, welfare and value deleteriously. Anomalous lower jaw can be curved downward (LJD), shortened (SJ) or misaligned (MA). Two separate groups of triploid Atlantic salmon (~12 g) with either normal lower jaw (NOR) or SJ were visually assessed four times over three months for presence and concurrence of jaw anomalies (with severity classified) and opercular shortening to understand the relatedness of these anomalous developmental processes. The prevalence of jaw anomalies increased in both groups over time (NOR group - SJ, LJD and MA combined 0-24.5%; SJ group - LJD and MA combined 17-31%). SJ and LJD occurred both independently and concurrently whereas MA exclusively concurred with them. All three anomalies could be concurrent. Severity of both LJD and SJ increased in the SJ group only. Opercular shortening recovery was observed in both groups but at a slower rate in the SJ group. The SJ group specific growth rate (SGR) was significantly (P < 0.05) lower than the NOR group. This study demonstrated the concurrence of SJ, LJD and MA and showed possible deleterious consequences deriving from the conditions.

Skeletal anomaly assessment in diploid and triploid juvenile Atlantic salmon (Salmo salar L.) and the effect of temperature in freshwater.

Triploid Atlantic salmon tend to develop a higher prevalence of skeletal anomalies. This tendency may be exacerbated by an inadequate rearing temperature. Early juvenile all-female diploid and triploid Atlantic salmon were screened for skeletal anomalies in consecutive experiments to include two size ranges: the first tested the effect of ploidy (0.2-8 g) and the second the effect of ploidy, temperature (14 °C and 18 °C) and their interaction (8-60 g). The first experiment showed that ploidy had no effect on skeletal anomaly prevalence. A high prevalence of opercular shortening was observed (average prevalence in both ploidies 85.8%) and short lower jaws were common (highest prevalence observed 11.3%). In the second experiment, ploidy, but not temperature, affected the prevalence of short lower jaw (diploids > triploids) and lower jaw deformity (triploids > diploids, highest prevalence observed 11.1% triploids and 2.7% diploids) with a trend indicating a possible developmental link between the two jaw anomalies in triploids. A radiological assessment (n = 240 individuals) showed that at both temperatures triploids had a significantly (P < 0.05) lower number of vertebrae and higher prevalence of deformed individuals. These findings (second experiment) suggest ploidy was more influential than temperature in this study.

A primary phosphorus-deficient skeletal phenotype in juvenile Atlantic salmon Salmo salar: the uncoupling of bone formation and mineralization.

To understand the effect of low dietary phosphorus (P) intake on the vertebral column of Atlantic salmon Salmo salar, a primary P deficiency was induced in post-smolts. The dietary P provision was reduced by 50% for a period of 10 weeks under controlled conditions. The animal's skeleton was subsequently analysed by radiology, histological examination, histochemical detection of minerals in bones and scales and chemical mineral analysis. This is the first account of how a primary P deficiency affects the skeleton in S. salar at the cellular and at the micro-anatomical level. Animals that received the P-deficient diet displayed known signs of P deficiency including reduced growth and soft, pliable opercula. Bone and scale mineral content decreased by c. 50%. On radiographs, vertebral bodies appear small, undersized and with enlarged intervertebral spaces. Contrary to the X-ray-based diagnosis, the histological examination revealed that vertebral bodies had a regular size and regular internal bone structures; intervertebral spaces were not enlarged. Bone matrix formation was continuous and uninterrupted, albeit without traces of mineralization. Likewise, scale growth continues with regular annuli formation, but new scale matrix remains without minerals. The 10 week long experiment generated a homogeneous osteomalacia of vertebral bodies without apparent induction of skeletal malformations. The experiment shows that bone formation and bone mineralization are, to a large degree, independent processes in the fish examined. Therefore, a deficit in mineralization must not be the only cause of the alterations of the vertebral bone structure observed in farmed S. salar. It is discussed how the observed uncoupling of bone formation and mineralization helps to better diagnose, understand and prevent P deficiency-related malformations in farmed S. salar.

Sulphated glycosaminoglycans and proteoglycans in the developing vertebral column of juvenile Atlantic salmon (Salmo salar).

In the present study, the distribution of sulphated glycosaminoglycans (GAGs) in the developing vertebral column of Atlantic salmon (Salmo salar) at 700, 900, 1100 and 1400 d° was examined by light microscopy. The mineralization pattern was outlined by Alizarin red S and soft structures by Alcian blue. The temporal and spatial distribution patterns of different types of GAGs: chondroitin-4-sulphate/dermatan sulphate, chondroitin-6-sulphate, chondroitin-0-sulphate and keratan sulphate were addressed by immunohistochemistry using monoclonal antibodies against the different GAGs. The specific pattern obtained with the different antibodies suggests a unique role of the different GAG types in pattern formation and mineralization. In addition, the distribution of the different GAG types in normal and malformed vertebral columns from 15 g salmon was compared. A changed expression pattern of GAGs was found in the malformed vertebrae, indicating the involvement of these molecules during the pathogenesis. The molecular size of proteoglycans (PGs) in the vertebrae carrying GAGs was analysed with western blotting, and mRNA transcription of the PGs aggrecan, decorin, biglycan, fibromodulin and lumican by real-time qPCR. Our study reveals the importance of GAGs in development of vertebral column also in Atlantic salmon and indicates that a more comprehensive approach is necessary to completely understand the processes involved.

Toxicity of road deicing salt (NaCl) and copper (Cu) to fertilization and early developmental stages of Atlantic salmon (Salmo salar).

In many countries, salting of ice or snow covered roads may affect aquatic organisms in the catchment directly or indirectly by mobilization of toxic metals. We studied the toxicity of road deicing salt and copper (Cu) on the vulnerable early life stages of Atlantic salmon (Salmo salar), from fertilization till hatching. Controlled episodic exposure to road salt (≥ 5,000 mg/L) during fertilization resulted in reduced swelling and less percent egg survival. Exposure to Cu both during and post fertilization caused delayed hatching. Larval deformities were, however found as an additional effect, when eggs were exposed to high salt concentration (≥ 5,000 mg/L) mixed with Cu (10 µg Cu/L) during fertilization. Thus, it appears that the sensitivity of early developmental stages of Atlantic salmon increased when exposed to these stressors, and road salt application during spawning can pose threat to Atlantic salmon in water bodies receiving road runoff. The study gives insight on assessment and management of risks on Atlantic salmon population posed by road related hazardous chemicals.

[Abnormal development of gonads of dwarf females and low survival of their offspring as the cause of rarity of resident populations of Atlantic salmon (Salmo salar L.)].

Fecundity and mass of eggs, as well as survival of offspring, of hatchery-reared small (less than 406 g) females of Atlantic salmon are determined. The absolute fecundity and mass of egg of artificially reared fish are sufficiently less than that of anadromous spawners. Small females are characterized by delayed spawning; histological analysis has demonstrated significant asynchrony in oocyte development of some specimens. Survival of offspring of dwarf females at the early stages of development is lower than of anadromous females. Our results agree completely with the published data. It is suggested that dwarf females are not able to withstand intraspecific and interspecific competition because of low reproductive ability. This explains rarity of natural populations of dwarf (landlocked) forms of Atlantic salmon.

The effect of water temperature on vertebral deformities and vaccine-induced abdominal lesions in Atlantic salmon, Salmo salar L.

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.

Molecular pathology of vertebral deformities in hyperthermic Atlantic salmon (Salmo salar).

BACKGROUND: Hyperthermia has been shown in a number of organisms to induce developmental defects as a result of changes in cell proliferation, differentiation and gene expression. In spite of this, salmon aquaculture commonly uses high water temperature to speed up developmental rate in intensive production systems, resulting in an increased frequency of skeletal deformities. In order to study the molecular pathology of vertebral deformities, Atlantic salmon was subjected to hyperthermic conditions from fertilization until after the juvenile stage. RESULTS: Fish exposed to the high temperature regime showed a markedly higher growth rate and a significant higher percentage of deformities in the spinal column than fish reared at low temperatures. By analyzing phenotypically normal spinal columns from the two temperature regimes, we found that the increased risk of developing vertebral deformities was linked to an altered gene transcription. In particular, down-regulation of extracellular matrix (ECM) genes such as col1a1, osteocalcin, osteonectin and decorin, indicated that maturation and mineralization of osteoblasts were restrained. Moreover, histological staining and in situ hybridization visualized areas with distorted chondrocytes and an increased population of hypertrophic cells. These findings were further confirmed by an up-regulation of mef2c and col10a, genes involved in chondrocyte hypertrophy. CONCLUSION: The presented data strongly indicates that temperature induced fast growth is severely affecting gene transcription in osteoblasts and chondrocytes; hence change in the vertebral tissue structure and composition. A disrupted bone and cartilage production was detected, which most likely is involved in the higher rate of deformities developed in the high intensive group. Our results are of basic interest for bone metabolism and contribute to the understanding of the mechanisms involved in development of temperature induced vertebral pathology. The findings may further conduce to future molecular tools for assessing fish welfare in practical farming.

Morphological and molecular characterization of developing vertebral fusions using a teleost model.

BACKGROUND: Spinal disorders are a major cause of disability for humans and an important health problem for intensively farmed animals. Experiments have shown that vertebral deformities present a complex but comparable etiology across species. However, the underlying molecular mechanisms involved in bone deformities are still far from understood. To further explicate the mechanisms involved, we have examined the fundamental aspects of bone metabolism and pathogenesis of vertebral fusions in Atlantic salmon (Salmo salar). RESULTS: Experimentally, juvenile salmon were subjected to hyperthermic conditions where more than 28% developed fused vertebral bodies. To characterize the fusion process we analyzed an intermediate and a terminal stage of the pathology by using x-ray, histology, immunohistochemistry, real-time quantitative PCR and in situ hybridization. At early stage in the fusion process, disorganized and proliferating osteoblasts were prominent at the growth zones of the vertebral body endplates. PCNA positive cells further extended along the rims of fusing vertebral bodies. During the developing pathology, the marked border between the osteoblast growth zones and the chondrocytic areas connected to the arches became less distinct, as proliferating cells and chondrocytes blended through an intermediate zone. This cell proliferation appeared to be closely linked to fusion of opposing arch centra. During the fusion process a metaplastic shift appeared in the arch centra where cells in the intermediate zone between osteoblasts and chondrocytes co-expressed mixed signals of chondrogenic and osteogenic markers. A similar shift also occurred in the notochord where proliferating chordoblasts changed transcription profile from chondrogenic to also include osteogenic marker genes. In progressed fusions, arch centra and intervertebral space mineralized. CONCLUSION: Loss of cell integrity through cell proliferation and metaplastic shifts seem to be key events in the fusion process. The fusion process involves molecular regulation and cellular changes similar to those found in mammalian deformities, indicating that salmon is suitable for studying general bone development and to be a comparative model for spinal deformities.

Effects of skeletal deformities on swimming performance and recovery from exhaustive exercise in triploid Atlantic salmon.

The occurrence of spinal deformity in aquaculture can be considerable, and a high rate of deformity has been suggested in triploid smolts in Tasmania. However, the physiological performance of fish with skeletal deformities has not been addressed. The swimming performance and oxygen consumption of triploid Atlantic salmon smolts with either a vertebral fusion (platyspondyly) or multifocal scoliosis were compared to normal (non-deformed) triploid smolts. Fish with vertebral fusion attained swim speeds similar to normal fish, whereas scoliotic fish were unable to attain comparable swim speeds. Routine and maximum oxygen consumption was higher for deformed fish compared with normal fish, translating into apparent increased routine metabolic scope in vertebral fusion fish, and equivocal scope in scoliotic fish compared with normal controls. Deformed fish developed a lower excess post-exercise oxygen consumption compared to non-deformed fish, suggesting they are either incapable of sustained anaerobic activity or possess an increased recovery capacity. These data suggest that skeletal deformity has differential effects on swimming performance depending upon the type of deformity but imposes a significant metabolic cost on salmon smolts.

Association of spinal deformity and vaccine-induced abdominal lesions in harvest-sized Atlantic salmon, Salmo salar L.

Spinal deformities in Atlantic salmon, Salmo salar L., have been described as a disease of multifactorial origin for which vaccines and time of vaccination have been suggested as risk factors. A vaccine efficacy trial where spinal deformity became evident was continued by the observational study reported here. In the preharvest part of the study 17 months post-sea transfer, there was a prevalence of 11.3% spinal deformity, with deformities present only in one vaccine group indicating a strong vaccine involvement. At slaughter, the prevalence of spinal deformities was 11.7%, and deformed fish had only 62% of normal slaughter weight. Visual analogue scales (VAS) were used for continuous recordings of vaccine-induced abdominal lesions and deformity. A logistic regression model associating presence of spinal deformity with markers of abdominal lesions was developed. The odds ratio for spinal deformity was 5.7 (95% CI: 3.4-9.4) for each unit increase in adhesion score (0-6) and 4.9 (2.9-3.4) for each unit increase in melanin on abdominal organs (0-3). Lesions in the dorsal caudal part of the abdomen gave an odds ratio for spinal deformity of 2.2.

Heat shock during early somitogenesis induces caudal vertebral column defects in Atlantic salmon (Salmo salar).

In several terrestrial vertebrates, heat shock (HS) during somitogenesis causes vertebral deformities. To determine if vertebral deformities can occur due to sudden temperature changes during early development in fish, Atlantic salmon embryos were HS treated during somitogenesis. Ten months later these individuals displayed a high prevalence of caudal vertebral column condensations (27-34%). The defects were located caudally of the abdominal cavity, displaying an even distribution in this region independent of time of HS. To determine if HS disturbed vertebral development during somitogenesis, two genes coding for markers of skeletal development were identified, namely, the secreted protein Shh (Sashh) and the transcription factor Twist (Satwist). These proteins are involved in the proliferation and specification of presumptive skeletal cells (sclerotome) in vertebrates. The spatial expression pattern of sashh and satwist in salmon indicated a functional conservation of these proteins. Furthermore, HS embryos displayed expressional disturbance in both sashh and satwist, indicating an effect of HS on sclerotomal cell patterning. However, the HS-protecting ability in embryos seems to be individually regulated because reduction in gene expression was not detected at all stages; in addition, HS did not induce somitic disturbance and vertebral deformity in all embryos.

Teratogenicity of elevated egg incubation temperature and egg vitamin A status in Atlantic salmon, Salmo salar L.

The present study was undertaken to investigate the possibility that high egg vitamin A (VA) status in combination with elevated egg incubation temperatures may cause deformities in Atlantic salmon, Salmo salar L. Egg batches selected for their total VA concentration were exposed to low (normal, 8 degrees C) or elevated (14 degrees C) egg incubation temperatures. Temperature was the main factor causing bone deformities such as warped gill opercula, fin and jaw deformities, but not for the development of spinal deformities where all groups displayed a 'baseline' occurrence of mild deformity (decreased vertebral size in the cephalic region) and no systematic variation in the occurrence of serious spinal deformities (fused vertebrae). A possible effect of egg incubation temperature fluctuation was found for the groups reared at low temperatures. An indication of a negative effect of elevated egg VA status for the development of organ deformities such as missing septum transversum and situs inversus was found in addition to temperature effects, however, no firm conclusions could be drawn from the present data. The phenotypes for temperature-induced deformities resembled the phenotype of VA-induced deformities, but no clear conclusions on the causality of the deformities found in the present study could be drawn. Egg incubation temperatures, both absolute temperature and temperature variations, should therefore be strictly controlled.