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Title: The ontogeny of osmoregulation in the Nile tilapia (Oreochromis niloticus L.)
Author: Fridman, Sophie
ISNI:       0000 0004 2701 7830
Awarding Body: University of Stirling
Current Institution: University of Stirling
Date of Award: 2011
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In recent times, diminishing freshwater resources, due to the rapidly increasing drain of urban, industrial and agricultural activities in combination with the impact of climate change, has led to an urgent need to manage marine and brackish water environments more efficiently. Therefore the diversification of aquacultural practices, either by the introduction of new candidate species or by the adaptation of culture methods for existing species, is vital at a time when innovation and adaptability of the aquaculture industry is fundamental in order to maintain its sustainability. The Nile tilapia (Oreochromis niloticus, Linnaeus, 1758), which has now been spread well beyond its natural range, dominates tilapia aquaculture because of its adaptability and fast growth rate. Although not considered to be amongst the most salt tolerant of the cultured tilapia species, the Nile tilapia still offers considerable potential for culture in low-salinity water. An increase in knowledge of the limits and basis of salinity tolerance of Nile tilapia during the sensitive early life stages and the ability to predict responses of critical life-history stages to environmental change could prove invaluable in improving larval rearing techniques and extend the scope of this globally important fish species. The capability of early life stages of the Nile tilapia to withstand variations in salinity is due to their ability to osmoregulate, therefore the ontogeny of osmoregulation in the Nile tilapia was studied from spawning to yolk-sac absorption after exposure to different experimental conditions ranging from freshwater to 25 ppt. Eggs were able to withstand elevated rearing salinities up to 20 ppt, but transfer to 25 ppt induced 100% mortality by 48 h post-fertilisation. At all stages embryos and larvae hyper-regulated at lower salinities and hypo-regulated at higher salinities, relative to the salinity of the external media. Osmoregulatory capacity increased during development and from 2 days post-hatch onwards remained constant until yolk-sac absorption. Adjustments to larval osmolality, following abrupt transfer from freshwater to experimental salinities (12.5 and 20 ppt), appeared to follow a pattern of crisis and regulation, with whole-body osmolality for larvae stabilising at c. 48 h post-transfer for all treatments, regardless of age at time of transfer. Age at transfer to experimental salinities (7.5 – 20 ppt) had a significant positive effect on larval ability to osmoregulate; larvae transferred at 8 dph maintained a more constant range of whole body osmolality over the experimental salinities tested than larvae at hatch. Concomitantly, survival following transfer to experimental salinities increased with age. There was a significant effect (GLM; p < 0.05) of salinity of incubation and rearing media on the incidence of gross larval malformation that was seen to decline over the developmental period studied. It is well established that salinity exerts a strong influence on development and growth in early life stages of fishes therefore the effects of varying low salinities (0 - 25 ppt) on hatchability, survival, growth and energetic parameters were examined in the Nile tilapia during early life stages. Salinity up to 20 ppt was tolerable, although reduced hatching rates at 15 and 20 ppt suggest that these salinites may be less than optimal. Optimum timing of transfer of eggs from freshwater to elevated salinities was 3-4 h post-fertilisation, following manual stripping and fertilisation of eggs, however increasing incubation salinity lengthened the time taken to hatch. Salinity was related to dry body weight, with larvae in salinities greater than 15 ppt displaying, at hatch, a significantly (GLM: p < 0.05) lower body weight but containing greater yolk reserves than those in freshwater or lower salinities. Survival at yolk-sac absorption displayed a significant (GLM; p < 0.05) inverse relationship with increasing salinity and mortalities were particularly heavy in the higher salinities of 15, 20 and 25 ppt. Mortalities occurred primarily during early yolk-sac development yet stabilised from 5 dph onwards. Salinity had a negative effect on yolk absorption efficiency (YAE). Salinity-related differences in oxygen consumption rates were not detectable until 3 days post-hatch; oxygen consumption rates of larvae in freshwater between days 3 – 6 post-hatch were always significantly higher (GLM p < 0.05) than those in 7.5, 15, 20 and 25 ppt, however, on day 9 post-hatch this pattern was reversed and freshwater larvae had a significantly lower QO2 than those in elevated salinities. Salinity had a significant inverse effect on larval standard length, with elevated salinities producing shorter larvae from hatch until 6 dph, after which time there was no significant differences between treatments. Salinity had a significant effect on whole larval dry weight, with heavier larvae in elevated salinities throughout the yolk-sac period (GLM; p < 0.05). The ability of the Nile tilapia to withstand elevated salinity during early life stages is due to morphological and ultrastructural modifications of extrabranchial mitochondria-rich cells (MRCs) that confer an osmoregulatory capacity before the development of the adult osmoregulatory system. A clearly defined temporal staging of the appearance of these adaptive mechanisms, conferring ability to cope with varying environmental conditions during early development, was evident. Ontogenetic changes in MRC location, 2-dimensional surface area, density and general morphological changes were investigated in larvae incubated and reared in freshwater and brackish water (15 ppt) from hatch until yolk-sac absorption using Na+/K+-ATPase immunohistochemistry with a combination of microscope techniques. The pattern of MRC distribution was seen to change during development under both treatments, with cell density decreasing significantly on the body from hatch to 7 days post-hatch, but appearing on the inner opercular area at 3 days post-hatch and increasing significantly (GLM; p < 0.05) thereafter. Mitochondria-rich cells were always significantly (GLM; p < 0.05) denser in freshwater than in brackish water maintained larvae. In both freshwater and brackish water, MRCs located on the outer operculum and tail showed a marked increase in size with age, however, cells located on the abdominal epithelium of the yolk-sac and the inner operculum showed a significant decrease in size (GLM; p < 0.05) over time. Mitochondria-rich cells from brackish water maintained larvae from 1 day post-hatch onwards were always significantly larger (GLM; p < 0.05) than those maintained in freshwater. Preliminary scanning electron microscopy studies revealed structural differences in chloride cell morphology that varied according to environmental conditions. Mitochondria-rich cell morphology and function are intricately related and the plasticity or adaptive response of this cell to environmental changes is vital in preserving physiological homeostasis and contributes to fishes’ ability to inhabit diverse environments. Yolk-sac larvae were transferred from freshwater at 3 days post-hatch to 12.5 and 20 ppt and sampled at 24 and 48 h post-transfer. The use of scanning electron microscopy allowed a quantification of MRC, based on the appearance and surface area of their apical crypts, resulting in a reclassification of ‘sub-types’ i.e. Type I or absorptive, degenerating form (surface area range 5.2 – 19.6 μm2), Type II or active absorptive form (surface area range 1.1 – 15.7 μm2), Type III or differentiating form (surface area range 0.08 – 4.6 μm2) and Type IV or active secreting form (surface area range 4.1 – 11.7 μm2). In addition, the crypts of mucous cells were discriminated from those of MRCs based on the presence of globular extensions and similarly quantified.
Supervisor: Rana, Krishen Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available
Keywords: Nile tilapia ; osmoregulation ; mitochondria rich cells ; chloride cells ; larvae ; Oreochromis niloticus ; adaptability ; salinity tolerance ; Nile tilapia ; Osmoregulation ; Sustainable aquaculture