Juvenile zebrafish (Danio rerio) are able to recover from lordosis.

Haemal lordosis, a frequent skeletal deformity in teleost fish, has long been correlated with increased mechanical loads induced by swimming activity. In the present study, we examine whether juvenile zebrafish can recover from haemal lordosis and explore the musculoskeletal mechanisms involved. Juveniles were subjected to a swimming challenge test (SCT) that induced severe haemal lordosis in 49% of the animals and then immediately transferred them to 0.0 total body lengths (TL) per second of water velocity for a week. The recovery from lordosis was examined by means of whole mount staining, histology and gene expression analysis. Results demonstrate that 80% of the lordotic zebrafish are capable of internal and external recovery within a week after the SCT. Recovered individuals presented normal shape of the vertebral centra, maintaining though distorted internal tissue organization. Through the transcriptomic analysis of the affected haemal regions, several processes related to chromosome organization, DNA replication, circadian clock and transcription regulation were enriched within genes significantly regulated behind this musculoskeletal recovery procedure. Genes especially involved in adipogenesis, bone remodeling and muscular regeneration were regulated. A remodeling tissue-repair hypothesis behind haemal lordosis recovery is raised. Limitations and future possibilities for zebrafish as a model organism to clarify mechanically driven musculoskeletal changes are discussed.

Skeletal ontogeny of the vertebral column and of the fins in shi drum Umbrina cirrosa (Teleostei: Perciformes: Sciaenidae), a new candidate species for aquaculture.

The osteological development of the vertebral column and fins in shi drum Umbrina cirrosa was studied in order to improve knowledge for its introduction in Mediterranean aquaculture. The osteological development was studied in 171 individuals, of total length (LT ) from 2·7 to 30·2 mm that were reared under the mesocosm technique. Vertebral ontogeny starts at 3·4 and 4·0 mm LT , with the formation of the first cartilaginous neural and haemal arches, and spines, respectively, and is completed with the full attainment of epicentrals (12·5 mm LT ). The formation of vertebral centra occurs between 4·1 and 7·4 mm LT . Pectoral supports are the first fin elements to develop (3·0 mm LT ), followed by those of the caudal fin (3·8 mm LT ), pelvic fin (3·9 mm LT ) and finally by those of the dorsal and anal fins (4·5 mm LT ). The caudal fin is the first to develop fin rays and attain the full count of principal fin rays (4·5-6·8 mm LT ), but the last to be fully completed with the formation of procurrent fin rays (6·9-17·5 mm LT ). The next fins starting to present rays are the dorsal (5·3 mm LT ) and the pectoral fins (5·6 mm LT ), while the anal and pelvic fins are the last (5·7 mm LT ). Following the caudal principal fin rays (6·8 mm LT ), the dorsal, anal (6·9 mm LT ), pelvic (7·4 mm LT ) and pectoral fins (9·8 mm LT ) are the next with fully completed ray counts. Aggregation of qualitative changes, such as the appearance of cartilages, the beginning and the complement of the ossification process and the full complement of elements in U. cirrosa were measured as cumulative frequency counts. These measurements reveal three ontogenetic intervals: one very developmentally active period during early life stages (from 3 to 5·9 mm LT ), a second slower developmental period (from 6·0 to 8·9 mm LT ) and finally a period of ontogeny more focused on structure refinement up to metamorphosis and settlement (>9·0 mm LT ).

Saddleback syndrome in European sea bass Dicentrarchus labrax (Linnaeus, 1758): anatomy, ontogeny and correlation with lateral-line, anal and pelvic fin abnormalities.

This study focused on the anatomy and ontogeny of saddleback syndrome (SBS) in reared European sea bass. The abnormality was detected at an unusually high frequency (12-94%) during a routine quality control in a commercial hatchery. Anatomically the abnormality was mainly characterized by the loss of 1-5 hard spines and severe abnormalities of the proximal pterygiophores (anterior dorsal fin), size reduction of a few lepidotrichia, missing lepidotrichia and/or lepidotrichia of poor ossification (posterior dorsal fin). SBS was significantly correlated with abnormalities of the anal and pelvic fins in all the examined populations. Moreover, in juvenile fish, SBS was significantly correlated with an abnormal lack of sectors of the lateral line. The examination of early larval samples revealed that SBS was ontogenetically associated with severe abnormalities of the primordial marginal finfold, which developed at the flexion stage (ca 8.5-11.0 mm SL). Histologically, these abnormalities were associated with extensive epidermal erosion. The results are discussed in respect of the critical ontogenetic period and the possible causative factors of SBS in European sea bass. It is suggested that causative factors acted during a wide ontogenetic period including flexion and metamorphosis phases.

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.

Thermally induced phenotypic plasticity of swimming performance in European sea bass Dicentrarchus labrax juveniles.

The vulnerability of embryonic and larval stages of European sea bass Dicentrarchus labrax to environmental temperature and the longer-term consequences for the early juveniles was demonstrated. This phenotypic plasticity was highlighted by subjecting D. labrax at 15.2 +/- 0.3 or 20.0 +/- 0.4 degrees C (mean +/-s.d.) up to metamorphosis and then at the same temperature (18.5 +/- 0.7 degrees C). After 4-6 weeks at the same temperature, the measurement of critical swimming speed at four exercise temperatures (15, 20, 25 and 28 degrees C) showed a significantly higher swimming capacity in the fish initially reared at 15 degrees C than for fish initially reared at 20 degrees C. This performance was correlated with significant differences in the phenotype of red muscle. Thermally induced phenotypic plasticity was clearly demonstrated as an important mechanism controlling swimming performance in early juveniles of D. labrax.

Temperature and myogenic factor transcript levels during early development determines muscle growth potential in rainbow trout (Oncorhynchus mykiss) and sea bass (Dicentrarchus labrax).

The influence of changes in environmental temperature on the mRNA levels of myogenic regulatory factors (MRFs), i.e. MyoD and myogenin, as well as myosin heavy chain (MyHC) were studied during early larval development in rainbow trout and sea bass. Phosphoimager analysis of northern blots indicated that there is an optimum temperature for the RNA transcript levels of MRF and MyHC RNA in trout and in sea bass larvae. In the trout strain studied, the highest concentration for MRF and MyHC transcripts was found at 8 degrees C rather than 4 degrees C or 20 degrees C. In European sea bass, the highest concentrations of MRF and MyHC mRNA were observed at 15-20 degrees C rather than 13 degrees C. Raising sea bass larvae at 15 degrees C was associated with higher MyHC gene expression as well as a trend towards an increase in total muscle fibre number and higher growth rates after transfer at ambient temperature. Results suggest that mRNA levels of MRF and MyHC can be used to optimise early development. An experiment in which the temperature was changed illustrates the consequence of precise temporal expression of MRF genes in specifying muscle fibre number at critical stages during early development.

Evidence of temperature-dependent sex determination in the European sea bass (Dicentrarchus labrax L.).

To test the hypothesis that sex determination in the European sea bass (Dicentrarchus labrax L.) Can be affected by the incubating temperature during the very early developmental stages, eggs from the same batch of spontaneously spawned broodstock were divided at the stage of half-epiboly into three groups according to rearing temperature: G13 = 13 degrees C, G15 = 15 degrees C, and G20 = 20 degrees C. Temperature treatment lasted until the middle of metamorphosis (17-18 mm total length, [TL]), and, with the exclusion of water temperature, all biotic and abiotic conditions were identical for the three experimental groups. The on-growing phase was performed under ambient photoperiod and temperature conditions for all groups. Sex proportions were determined by histological examination of the gonads of fish at 308, 467, and 568 days posthatch (DPH). At 308 DPH (TL: 135-201 mm), 100% of the specimens had differentiated into males and females. A significantly higher (P < 0.01) proportion of females was found in groups G13 (72-74%) and G15 (67-73%) than in group G20 (24-28%). At the final sampling there was no statistically significant difference in body weight between the experimental groups. However, in all groups, female fish were larger than males (P < 0.001). Results provide for the first time clear evidence that temperature during the very early developmental stages is the crucial factor affecting the process of sex differentiation of the sea bass, with low rearing temperatures (13 or 15 degrees C) resulting in sex proportions consistently skewed in favor of females.