Rainbow trout (Oncorhynchus mykiss) were exposed to ion-poor (soft) water to test the hypothesis that naturally induced proliferation of branchial chloride cells causes a thickening of the blood-to-water diffusion barrier. This was achieved by using a combination of scanning and transmission electron-microscopic techniques. Fish were exposed to soft-water conditions ([Na+]= 0.055 mmol l-1, [Cl-] approximately 0.029 mmol 1(-1), [Ca2+] approximately 0.059 mmol 1(-1), and [K+] approximately 0.007 mmol 1(-1)) for 1, 2, and 4 weeks. Marked chloride cell proliferation was evident at all sampling times with an approximate doubling of the gill epithelial surface area covered by chloride cells exposed to the water ("chloride cell fractional area"). The increases in chloride cell fractional area resulted from both increased numbers of cells and expanded apical surfaces of exposed individual cells. As a result of chloride cell proliferation, soft-water exposure was associated with a doubling of the lamellar blood-to-water diffusion distance from 3.26+/-0.08 microM to 6.58+/-0.43 microM as determined from transmission electron micrographs. These data demonstrated a positive correlation between chloride cell fractional area and blood-to-water diffusion distance. We conclude that, in trout, chloride cell proliferation during soft-water exposure, while presumably benefiting ionic regulation, may impair gas transfer owing to the associated thickening of the blood-to-water diffusion barrier.
Radiotracer techniques were used to measure influx and efflux rates of Ca2+ in freshwater-adapted Oreochromis mossambicus. The influx rate of Ca2+ is related to body weight (W) as Fin = 50W0.805 nmol Ca2+/h. For a 20-g fish the calculated influx rate was 558 nmol Ca2+/h, and this was attributed largely to extraintestinal uptake since the drinking rate was estimated to be only 28 microliter water/h, which corresponds to an intake of 22.4 nmol Ca2+/h. The Ca2+ efflux rate was calculated using the initial rate of appearance of radiotracer in the ambient water and the specific activity of plasma Ca2+. Tracer efflux rates were constant over 6-8 h, which indicated that there was no substantial loss of tracer in either the urine or the feces because this would have resulted in random bursts of tracer loss. Efflux rates then primarily represent integumentary and presumably branchial efflux rates. The efflux rate of Ca2+ is related to body weight as Fout = 30W0.563 nmol Ca2+/h, which means an efflux rate of 162 nmol Ca2+/h for a 20-g fish. The net whole-body Ca2+ influx, calculated as Fnet = Fin - Fout, was 396 nmol/h for a 20-g fish, which proves that the ambient water is an important source of Ca2+.
The involvement of the freshwater fish gill chloride cells (CCs) in trans-branchial calcium uptake (JinCa(2+)) was investigated. This was accomplished by assessing the interspecific relationships between the apical surface area of CCs exposed to the external environment and JinCa(2+). Three species of freshwater teleosts, the rainbow trout (Oncorhynchus mykiss), the American eel (Anguilla rostrata) and the brown bullhead catfish (Ictalurus nebulosus), were used. Chronic (ten-day) treatment with cortisol in each species was used as a tool to evoke variations in both JinCa(2+) and gill CC morphology in order to assess intraspecific relationships between CC surface area and JinCa(2+). The results of quantitative morphometry, based on analysis of scanning electron micrographs, demonstrated that catfish possessed the lowest fractional area of exposed CC (CCFA) on the gill filament epithelium (12,744 ± 2248 μm(2)/mm(2)) and was followed, in increasing order, by American eel (21,355 ± 981 μm(2)/mm(2)) and rainbow trout (149,928 ± 26,545 μm(2)/mm(2)). With the exception of catfish, chronic treatment with cortisol caused significant increases in CCFA owing to proliferation of CCs and/or enlargement of individual CCs (eel only). The rates of JinCa(2+) closely reflected the CC fractional area in each species. The results of correlation analysis revealed significant correlations between CC fractional area and JinCa(2+) in trout and eel. Owing to the absence of an effect of cortisol treatment, there was no significant correlation in catfish because of insufficient variation in CC fractional area in this species. CC fractional area was significantly correlated with JinCa(2+) among the three species examined. These results suggest that CC is involved in calcium uptake in freshwater teleosts and that both intra- and interspecific differences in the rates of calcium uptake can be accounted for by variability in the surface area of exposed CCs on the gill epithelia.
Atlantic salmon (Salmo salar) parr were subjected to one of two photoperiod regimens: a simulated natural photoperiod, or a reciprocal photoperiod (with a light-to-dark ratio opposite to that of the natural photoperiod). The animals were held at 10 °C from mid-December onward and were sampled at 3-week intervals.By February, salmon held under the reciprocal photoperiod were significantly longer, had lower condition factors, and looked and behaved more like smolts than did those under the natural photoperiod. A separate experiment showed that salinity tolerance increased at this time. In June the situation was reversed; the salmon kept under the natural photoperiod showed greatest growth and high salinity tolerance.Longer days in either regimen coincided with an increase in number and apparent activity of pituitary somatotrops of the animals. Adrenocorticotropin (ACTH), pars intermedia, and prolactin cells were also judged to be active under these conditions. A role for growth hormone and other peptides as components of a 'light–pituitary axis' is discussed in relation to the parr–smolt transformation of Atlantic salmon.
Direct impact of ambient (1.95 W/m2) and subambient doses of UV-B radiation on muscle/skin tissue antioxidant status was assessed in mature zebrafish (Brachydanio rerio). The influence of these doses on hatching success and survival in earlier life stages was also examined. Subambient doses of UV-B radiation in the presence (1.28 W/m2) and absence (1.72 W/m2) of a cellulose acetate filter significantly depressed muscle/skin total glutathione (TGSH) levels compared with controls (0.15 W/m2) and low (0.19 W/m2) UV-B-treated fish after 6 and 12 h cumulative exposure. Ambient UV-B exposure significantly decreased muscle/skin glutathione peroxidase (GPx) activity after a 6 h exposure; activities of glutathione reductase (GR) were unchanged over this exposure period. Superoxide dismutase (SOD) and catalase activities peaked after 6 and 12 h cumulative exposure, respectively, but fell back to control levels by the end of the exposure period. The changes in tissue antioxidant status suggested UV-B-mediated increases in cytosolic superoxide anion radicals (O2-) and hydrogen peroxide (H2O2). This apparent UV-B-mediated increase in oxidative stress is further supported by a significant increase in muscle/skin thiobarbituric acid reactive substances (TBARS). Hatching success of newly fertilized eggs continuously exposed to ambient UV-B was only 2% of the control value. Even at 30 and 50% of ambient UV-B, hatching success was only 80 and 20%, respectively, of the control. Newly hatched larvae exposed to an ambient dose of UV-B, experienced 100% mortality after a 12 h cumulative exposure period. This study supports a major impact of UV-B on both the mature and embryonic zebrafish.
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