Three different levels of dietary vitamin D3 fed to first-feeding fry of Atlantic salmon (Salmo salar L.): effect on growth, mortality, calcium content and bone formation

Atlantic salmon ( Salmo salar L.) fry were reared on a fishmeal-based diet with three different levels of vitamin D₃ (0.2, 5 and 57 mg vitamin D₃ kg^–1 feed, ww) from first-feeding for 14 weeks. No significant differences were recorded in weight, length, specific growth rate, mortality, or kidney calcium concentration between the different dietary groups. No skeletal malformations or histopathological changes were recorded in any of the dietary groups. These results suggest Atlantic salmon fry to be highly tolerant of megadoses of vitamin D₃ over a limited period of time.     

养殖军曹鱼稚鱼骨骼畸形研究

本研究采用软骨—硬骨双染色技术对常规养殖条件下的军曹鱼(Rachycentron canadum)稚鱼全骨骼进行染色,观察并分析其骨骼畸形发生部位及相应的畸形类型。结果显示,在同一批次繁育的180个军曹鱼稚鱼(25日龄)骨骼标本中,有72个标本存在畸形情况,畸形率为40.00%。骨骼畸形类型共计22种,畸形率由高到低主要表现为米克尔氏软骨畸形、尾上骨缺失、脉棘分叉、基舌骨异位和尾上骨愈合等;所有骨骼畸形均未表现出显著可见的外部形态变化。但正常个体与骨骼畸形个体的全长存在极显著差异(P<0.01)。本研究表明,骨骼畸形对军曹鱼生长产生了影响。本研究为探索鱼类骨骼畸形的发生过程和原因、减少畸形率和优化苗种培育养殖条件提供了理论基础。     

Dietary fatty acids and inflammation in the vertebral column of Atlantic salmon, Salmo salar L., smolts: a possible link to spinal deformities

Vegetable oils (Vo) are an alternative to fish oil (Fo) in aquaculture feeds. This study aimed to evaluate the effect of dietary soybean oil (Vo diet), rich in linoleic acid, and of dietary fish oil (Fo diet) on the development of spinal deformities under bacterial lipopolysaccharide (LPS)‐induced chronic inflammation conditions in Atlantic salmon, Salmo salar L. Fish [25 g body weight (BW)] were fed the experimental diets for 99 days. On day 47 of feeding (40 g BW), fish were subjected to four experimental regimes: (i) intramuscular injections with LPS, (ii) sham‐injected phosphate‐buffered saline (PBS), (iii) intraperitoneally injected commercial oil adjuvant vaccine, or (iv) no treatment. The fish continued under a common feeding regime in sea water for 165 more days. Body weight was temporarily higher in the Vo group than in the Fo group prior to immunization and was also affected by the type of immunization. At the end of the trial, no differences were seen between the dietary groups. The overall prevalence of spinal deformities was approximately 14% at the end of the experiment. The Vo diet affected vertebral shape but did not induce spinal deformities. In groups injected with LPS and PBS, spinal deformities ranged between 21% and 38%, diet independent. Deformed vertebrae were located at or in proximity to the injection point. Assessment of inflammatory markers revealed high levels of plasma prostaglandin E₂ (PGE₂) in the Vo‐fed and LPS‐injected groups, suggesting an inflammatory response to LPS. Cyclooxigenase 2 (COX‐2) mRNA expression in bone was higher in fish fed Fo compared to Vo‐fed fish. Gene expression of immunoglobulin M (IgM) was up‐regulated in bone of all LPS‐injected groups irrespective of dietary oil. In conclusion, the study suggests that Vo is not a risk factor for the development of inflammation‐related spinal deformities. At the same time, we found evidence that localized injection‐related processes could trigger the development of vertebral body malformations.     

Long-term culture of Atlantic salmon (Salmo salar L.) in submerged cages during winter affects behaviour, growth and condition

In the search for alternative farming methods, we investigated whether large salmon submerged below 10 m in winter conditions behaved normally and performed as well as control fish held in standard surface cages. On average, 2345 salmon of ~ 3.5 kg were kept in each of six 2000 m³ sea-cages for 6 weeks; three of which were submerged to 10–24 m depth and three acted as surface controls (0–14 m). Behaviour during both day and night was studied with echo-sounders, and underwater video cameras fitted with infra-red lamps. A sub-sample of fish from each cage was weighed, measured and assessed for fin and snout condition prior to and after the experimental period. In addition, the vertebral column of 50 fish from the control and submerged treatments were dissected and X-rayed to assess vertebral deformities. The submerged salmon seemed unable to re-fill any gas into the swim bladder, as a linear decrease in echo reflection to < 5% of pre-submergence levels after 22 days of submergence indicated loss of almost all gas from the physostomous swim bladders and negatively buoyant fish. Around day 22, submerged salmon swam at night time with a distinct ‘tail-down, head-up’ tilt (26°) compared to the horizontal swimming position of control fish (− 3°). Average swimming speed (body length per second) of submerged salmon were 1.3–1.4 times faster (day: 0.77 ± 0.02; night: 0.46 ± 0.02, (mean ± SE)) than control fish (day: 0.54 ± 0.01; night: 0.37 ± 0.02) both during day and night. Almost no mortality was seen, and the submerged salmon maintained similar diurnal vertical migrations as the surface fish, indicating that deep submergence did not exhaust the fish. However, submerged fish fed less efficiently, resulting in lower growth and reduced feed utilization. Fins and snouts of the submerged fish had small, but significantly more erosion than the control fish. Vertebrae in the tail region were significantly compressed in the submerged fish compared to control fish. This could be an early symptom of development of vertebral deformities. The results suggest that continuous submergence below 10 m for longer than 2 weeks reduces the welfare and performance of Atlantic salmon.     

Phenotypic effects of antibiotic‐induced axenity and egg disinfection in early larval European seabass (Dicentrarchus labrax L.)

In the present study, the hypothesis that the difference in axenic conditions in the incubation and rearing environment of European seabass larvae induces size and shape effects on the specimens is tested. This difference is studied between xenic and axenic seabass larvae of DAH (day after hatching) 0, 5, 11 and 15. The axenic rearing protocol involves an egg disinfection with glutaraldehyde after the primary one with iodine in the hatchery, and the hypothesis that this secondary disinfection induces size and shape effects is also tested. In order to accomplish this, three egg and larvae treatments are included: “DA” (disinfected axenic), “DX” (disinfected xenic) and “NX” (nondisinfected xenic). Regarding the effect of antibiotic‐induced axenity, DA larvae exhibited larger bodies than both DX and NX on DAH 5 and 11. They also had a smaller yolk sac than DX at hatching, but consumed it slower. Towards the end of the experiment, DA larvae were thicker, but slightly more curved than DX and NX, which may be an abnormal shape, or a slightly more advanced ontogenetic stage. As far as egg disinfection, it had significant but very moderate shape effects on DAH 5 and 11, and disinfected larvae consumed their yolk sac faster. To the best of our knowledge, this study is one of the first to illustrate the subtle but significant size and shape effects caused by antibiotic‐induced axenity and secondary egg disinfection in the early larval stages, which suggest the existence of bacterial mechanisms that play a phenotypic role.     

Dietary phosphorus does not reduce the risk for spinal deformities in a model of adjuvant‐induced inflammation in Atlantic salmon (Salmo salar) postsmolts

Inflammation is a non‐specific protective mechanism towards injury known to affect bone remodelling. This study aimed to investigate the effect of Freund’s complete adjuvant (FCA) induced‐inflammation on the prevalence of spinal deformities of Atlantic salmon postsmolts fed with two different dietary P levels. Sextuple groups of salmon postsmolts were fed with either a low‐phosphorous (6 g kg^?1 available P, LP) or a high‐phosphorous (9 g kg^?1 available P) diet for a period of 101 days. On Day 102, individually tagged fish were subjected to (i) single injection with FCA (0.125 mg kg^?1 BW) dissolved in phosphate‐buffered saline (PBS) (ii) placebo injection with PBS or (iii) sham injection (insertion of needle only) or (iv) remained untreated. On Day 103, fish were given a common diet for 174 days in seawater. No significant differences in body weight were observed. Injected fish, particularly the FCA group, had more compressions in the injection site than untreated fish. No effect of diet and no interaction between treatment and diet were observed. Severe scoliosis was observed in ～7% of FCA‐injected individuals, corresponding to a mixture of bone malformations in the tail region. In conclusion, experimentally induced inflammation may be an independent risk factor for bone deformities in Atlantic salmon.     

A new type of lordosis and vertebral body compression in Gilthead sea bream,Sparus aurata L.: aetiology,anatomy and consequences for survival

A new type of vertebral malformation is described, consisting of deformed cartilaginous neural and haemal processes and the compression and fusion of vertebral bodies. The malformation is designated as haemal vertebral compression and fusion (haemal VCF). We studied the aetiology of the malformations and described microanatomical histopathological alterations. The malformations were detected during routine quality control in one of six monitored Gilthead sea bream populations. Haemal VCF affected the posterior part of the vertebral column (haemal vertebrae). In 20% of the deformed specimens, haemal VCF was combined with lordosis. At 35 dph (days post‐hatching), early anatomical signs of the haemal VCF consisted of abnormal centrum mineralization, malformed cartilaginous neural and haemal processes and developing lordotic alterations. The histological examination of the deformed individuals revealed that haemal VCF is preceded by notochord abnormalities. The frequency of deformed individuals was three times higher at 35 than at 61 dph (50.3% vs. 17.2%, n = 157 and n = 250, respectively). No signs of repair or reversion of malformations have been observed. Thus, the steep decrease in deformities in older animals suggests that haemal VCF is linked to high mortality rates. The results are discussed in respect of the possible causative factors of haemal VCF.     

Short- and long-term differences in growth, feed conversion efficiency and deformities in juvenile Atlantic cod (Gadus morhua) startfed on rotifers or zooplankton

Growth, feed conversion efficiency and frequencies of skeletal deformities were studied in juvenile Atlantic cod (Gadus morhua) that had been startfed on either rotifers (rotifer group) or zooplankton (zooplankton group). After metamorphosis, the fish were reared at four constant temperatures (7, 10, 13, 16°C) or moved successively from 16 to 13 and 10°C (T‐step, average 13.2°C). The zooplankton group had a consistently higher growth rate at all the temperatures studied. Further, the zooplankton group had higher food intake (20%) and higher feed conversion ratio (1.65 vs. 1.31). In addition, a significantly higher incidence of skeletal deformities was found in the rotifer group (14.2%) compared with the zooplankton group (4.1%). After termination of the laboratory study, the fish were reared in sea pens under ambient conditions for 17 months. Final weights of the zooplankton group were consistently larger (between 12% and 14% larger depending on original temperature groups). To verify the growth results, we conducted a follow‐up study where a single egg group was divided into two parts and fed either on rotifers or zooplankton. This study indicated similar growth differences as found in the first study. Overall, our data suggest that different startfeeding diets may be important for long‐term growth, incidence of deformities and quality of juvenile cod. The use of zooplankton can greatly improve long‐term growth and quality of cod juveniles. The study also highlights the advantage of using elevated temperatures in the juvenile phase as this will lead to significantly higher final weights in the adult stage.     

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.     

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.