G. Ramírez‐Zarzosa

Temperature effects on muscle growth in two populations (Atlantic and Mediterranean) of sea bass, Dicentrarchus labrax L.

Two genetically different populations of sea bass, Dicentrarchus labrax L., Atlantic (Atl.) and Mediterranean (Med.), were subjected to the following incubation/cultivation temperatures (T): 15 °C/natural, 17 °C/natural, 15/19 °C, 17/19 °C (natural T averaging 15 °C and raising gradually). Muscle cellularity was measured at different larval stages for each T regime. During the vitelline phase, muscle growth was mainly due to muscle fibre hypertrophy. In Med. larvae, higher incubation T (17 °C) increased the area of white and red fibres at hatching, while in Atl. larvae there was no significant T effect at this stage. At mouth opening, the area of white fibres increased at 19 °C in Atl. larvae (P<0.05), but in Med. larvae it was similar for all temperatures. Following yolk-sac reabsorption, hypertrophy and hyperplasia increased in both populations. In these stages (20–55 days), both parameters were greater at 19 °C (P<0.05). Metamorphosis finished earlier at 19 °C. At this stage, Atl. larvae reared at 19 °C showed higher value of total myotomal area than at natural T, while in Med. sea bass, larvae reared at 19 °C showed a lower size of the myotome than at 17 °C/natural. In larvae from both populations reared at natural T, incubating T had a positive effect at the end of metamorphosis, thus the total myotomal area were higher at 17 °C/natural than at 15 °C/natural. Following metamorphosis, all groups showed a rapid growth, but higher at 19 °C (P<0.05). The results indicate that muscle cellularity was clearly influenced by T, and that both populations had different levels of response. These differences can be apparently associated with their respective genetic background.

Early temperature effects on muscle growth dynamics and histochemical profile of muscle fibres of sea bass Dicentrarchus labrax L., during larval and juvenile stages

Recently, it has been found that the thermal experience during the earliest phases of development could determine the larval and postlarval growth characteristics of teleosts. In order to investigate the effects of the early temperature regime on the advanced stages of growth of sea bass, Dicentrarchus labrax L., this species was reared during the vitelline phase at two temperatures: natural temperature (i15 jC) and 17.7F0.1 jC, and then larvae transferred to common temperature (natural temperature). Muscle growth was studied by morphometric and histochemical techniques (mATPase and NADH-TR). Body length and body mass were also measured. During the vitelline phase, muscle growth was similar in both experimental groups, but at 25 days, both hypertrophy and hyperplasia of white muscle fibres were greater in the prewarmed group (p<0.05). At the end of metamorphosis (80 days) and at 120 days, the average diameter of white muscle fibres, as well as the body length, were greater in the prewarmed group (p<0.05), but the number of white fibres did not differ significantly between groups. The morphological mosaic of white muscle fibres was observed at the end of metamorphosis, and the histochemical mosaic appeared gradually since the early postlarval stages. Thus, at 120 days, some specimens in both experimental groups showed three or four different mATPase staining white fibres: low (L), moderate (M), high (H) and/or very high (vH), whereas in other specimens, only L or M mATPase activity fibres were observed. Early T influenced the histochemical maturity of the

Muscle Development in Gilthead Sea Bream (Sparus aurata, L.) and Sea Bass (Dicentrarchus labrax, L.): Further Histochemical and Ultrastructural Aspects

The histochemical profiles ‐ mATPase and NADH‐TR reactions ‐ of the red and white muscle fibres of gilthead sea bream and sea bass were determined from the first week after hatching. Modifications of the mATPase technique by combinations of pH/time/molarity were carried out in order to compare the sensitivity of the myosin ATPase of each muscle fibre type of the lateral muscle. Results showed that the staining of muscle fibres was independent of small modifications in the technique. The intermediate ‘pink’ muscle was histochemically defined towards the end of the larval life and is considered to be implicated in the growth of the myotome. A layer of external cells was observed, by electron microscopical examination, between the connective tissue of the skin and the superficial red muscle fibres of larvae and postlarvae. It is suggested that the external cells are unlikely to be a source of red muscle fibres and implicated on the growth of the myotome, but rather a part of the dermatome. The timing, areas and mechanisms of hyperplastic growth of the myotome were defined and discussed.

The Post-larval Development of Lateral Musculature in Gilthead Sea Bream Sparus aurata (L.) and Sea Bass Dicentrarchus labrax (L.)

Fibre‐type differentiation of lateral musculature has been studied in gilthead sea bream Sparus aurata (L.) and sea bass Dicentrarchus labrax (L.) during post‐larval development using ultrastructural, histochemical and morphometric techniques. The study showed three muscle layers: red, intermediate (or pink) and white. Initially, most of the red muscle showed low myosin ATPase (m‐ATPase) activity fibres, whereas near the transverse septum some small high m‐ATPase activity fibres appeared and later acquired a rosette aspect. Afterwards, during adult growth the red muscle showed a histochemical mosaic appearance. The pink muscle in sea bass was observed at the beginning of juvenile development by the oxidative technique (NADH‐RT reaction) whereas in gilthead sea bream it was also observed at the end of larval development. The pink layer consists of high m‐ATPase activity fibres. However, along the muscle development other low and moderate m‐ATPase activity fibres were observed close to the red and white muscles, respectively. The white muscle of juvenile fish showed a histochemical mosaic appearance near the pink muscle. In adult specimens the mosaic white muscle spread out occupying the whole of the myotome. Morphometric analysis shows a significant increase in mean fibre diameter during post‐larval development, as shown by the Students t‐test (hypertrophic growth). Skewness and kurtosis values of fibre diameters point to the generation of a new fibres from the myosatellite cells (hyperplastic growth).

Early muscle injuries in a standard reared stock of sea bass Dicentrarchus labrax, (L.)

Abstract An early myopathy affecting the myotomes of a stock of sea bass is described. By electron microscopical examination different degrees of muscle lesions were first detected in recently hatched larvae. Muscle fibres underwent a degenerative process, that according to the gravity of the lesions and age of the fish was sequenced in four stages. Necrotic muscle fibres were abundant in the myotomes of post-larvae (more than 60 days). Although this degenerative myopathy is non-specific, the importance lies in its progressive sequence. Vitamin E-selenium deficiency and uncontrolled peroxidation of the lipid content in the feed is thought to be the cause of the problem. The possible implication of the myopathy in the appearance of a high percentage of lordosis in the same stock of sea bass is discussed.

Red muscle development of gilthead sea bream Sparus aurata (L.): structural and ultrastructural morphometry.

The transverse red and white muscle area, the superficial red muscle fibres area and their percentages of mitochondria (%mit), myofibrils (%myof) and sarcoplasm (%sarc) were determined in the Mediterranean teleost gilthead sea bream, Sparus aurata (L.). Fish aged from hatching to 78 days were studied. The proportional growth of the red and white muscles was higher for the red muscle in the first half of the larval stage (1–35 days). Then the opposite relationship was observed. The hypertrophic growth of the superficial muscle fibres was continuous except in the first week after hatching. The percentage of mitochondria and percentage of myofibrils showed a significant change just after the half of the larval stage. Whereas the %mit of the superficial muscle fibres was higher than %myof from the first week after hatching to 35 days (average 66.64 %), then the %mit decreased significantly and at 73–78 days both parameters were close to an average value of 50 %. The meaning of these morphological changes is discussed in relation to the functional role of the red muscle of larvae and the onset of the gills respiration.

Effect of thermal treatment on muscle tissue structure and ultrastructure of wild and farmed sea bass, Dicentrarchus labrax L

The present work studies the muscle tissue structure and ultrastructure in two populations: wild and farmed sea bass, Dicentrarchus labrax L., in both raw and cooked states. Results found in fresh raw muscle tissue ultrastructure of wild sea bass showed some typical early post-mortem alterations: fibre-to-fibre detachment, detachment of myofibrils to endomysium and swelling of mitochondria and sarcoplasmic reticulum, mainly. In cooked samples of wild and farmed groups, sarcolemma and endomysium were disrupted and coagulated in most of the zones. Myofibrils were detached from sarcolemma–endomysium. Electron-dense aggregates were observed in both subsarcolemmal and interstitial spaces. These granular aggregates also appeared close to sarcolemma and endomysium, like an electron-dense chain. The interstitial material was quantified in both groups, and it was more abundant in farmed than in wild specimens. The size of the cooked fibres was better preserved in wild than in farmed sea bass, which evidences a higher thermal resistance of muscle tissue in wild specimens.