Ubiquitous increase in taurine transporter mRNA in tissues of tilapia (Oreochromis mossambicus) during high-salinity adaptation

We have isolated a cDNA encoding the taurine transporter from a tilapia (Oreochromis mossambicus) gill cDNA library. Transient expression of the cDNA in COS-7 cell indicates that the clone encodes a Na⁺- and Cl^–-dependent and -amino acid-specific taurine transporter. By the transfer of tilapia cultured in freshwater to 70% artificial seawater, plasma osmolality increased by up to 100–135 mOsm/kgH₂O along with the marked increase in the taurine transporter mRNA level in all the tissues examined i.e., kidney, stomach, intestine, gill, eye, liver, fin, muscle and brain. In most tissues, time-dependent change in the taurine transporter mRNA level corresponds to that in plasma osmolality. However, fin showed an acute and muscle showed a delayed increase in taurine transporter mRNA compared to changes in plasma osmolality. The taurine transporter mRNA level in tilapia embryos also increased after transfer from freshwater to 100% artificial seawater. Increase in taurine transporter expression leads to the activation of cellular uptake of taurine from plasma and the accumulation of taurine in the cell. Thus the results in the present study suggest that taurine plays an important role as an osmolyte in the ubiquitous tissues of tilapia during high-salinity adaptation.     

The effect of hepatic passage on postprandial plasma lipid profile of rainbow trout (Oncorhynchus mykiss) after a single meal

For the first time, pre‐ and post‐hepatic plasma lipid profiles were monitored following a single meal in a free‐swimming, non‐anaesthetized fish. Rainbow trout (Oncorhynchus mykiss; 700–1500 g; 10 °C) were equipped with cannulae in the dorsal aorta (DA) and hepatic portal vein (HPV). Simultaneous blood samples, taken from both cannulae at 0, 3, 6, 12, 24 and 48 h postprandial, revealed the time course of the plasma lipid profiles following a single meal (1% of body mass). Primarily monounsaturated fatty acids with the exception of 18:1n ? 9, increased significantly from baseline by 12 h postprandial without greatly affecting total plasma lipid concentrations. Total plasma lipids then showed a small peak at 24 h postprandial, coinciding with a peak in triacylglycerols. We conclude that assimilation of lipids from the digest into the plasma is slower than reported for proteins and carbohydrates in the same species. Furthermore, as there were no significant differences between the HPV and DA, no measurable effect of hepatic passage on plasma lipid levels was resolved. Therefore, we also conclude that, in contrast to that in higher vertebrates, hepatic passage does not seem to have a major role in rainbow trout for modulating the postprandial plasma profile of lipids.     

Influence of pond fertilization and feeding rate on growth performance, economic returns and water quality in a small-scale cage-cum-pond integrated system for production of Nile tilapia (Oreochromis niloticus L.)

The effects of pond fertilization and feeding rate on growth, economic returns and water quality were investigated to develop a low‐cost cage‐cum‐pond integrated system for production of Oreochromis niloticus (L.). Hand‐sexed male fingerlings averaging 19±0.39 and 32±0.69 g were stocked in cages and open ponds at 150 fish cage^?1 and 2 fish m^?2 respectively. Fish were cultured for 114 days in five triplicate treatments. Cages were installed into ponds and caged fish were fed a 24% protein diet at 3% (T1) and 6% (T2) body weight day^?1 (BWD) without pond fertilization, and 6% BWD with pond fertilization (T3). The open water in the fourth treatment (T4) was not stocked but contained caged fish, which were fed 6% BWD for the first 57 days followed by 3% BWD for the remaining period. Ponds in the control (T5) had no cages and were neither fertilized nor open‐pond fish fed. Feeding rate and pond fertilization significantly (P<0.05) affected fish growth, profitability and water quality among treatments. Fish growth, feed utilization, fish yield, water quality and profits were significantly (P<0.05) better in T3 than the other treatments. It was concluded that fish production and economic returns were optimized at 6% BWD in fertilized ponds.     

Immune protection in carp, Cyprinus carpio L., after immunization with Aeromonas hydrophila crude lipopolysaccharide

Abstract. Vaccination with crude lipopolysaccharide (LPS) induced better protection against infection with Aeromonas hydrophila in carp than vaccination with formalin killed vaccine. Dipping fish in vaccine for 2 h at 25°C was more effective than intraperitoneal injection of the vaccine in procedural simplicity, lower stress loading and the degree of protection acquired. In carp immunized with crude LPS by the dip method, antibodies were not detected by bacteriai agglutination, passive haemagglutination and the agar diffusion tests. The results indicate that the protection against A. hydrophila infection in carp is not dependent on humoral immunity.     

Protective immunity of sevenband grouper, Epinephelus septemfasciatus Thunberg, against experimental viral nervous necrosis

This paper describes the protective immune responses of sevenband grouper, Epinephelus septemfasciatus Thunberg, immunized with live piscine nodavirus, the causative agent of viral nervous necrosis (VNN), or the Escherichia coli – expressed recombinant coat protein. Nodavirus-neutralizing antibodies were detected at titres ranging from 1:158 to 1:1257 in serum of sevenband grouper which survived intramuscular injection with the virus, by a cell culture assay system. The virus-neutralizing ability of immune serum was also confirmed by injecting virus previously treated with serum into fish. This indicates establishment of acquired immunity in survivors and thus explains why survivors from natural infection are resistant to recurrence of the disease. Young sevenband grouper were immunized twice by intramuscular injections with the recombinant coat protein. Immunized fish produced neutralizing antibodies at high titres for at least 110 days and showed significantly lower mortalities in virus challenge tests. These results suggest the potential for vaccination against VNN in sevenband grouper, which is susceptible to piscine nodavirus at all life-stages.     

Compensatory growth in Atlantic halibut: Effect of starvation and subsequent feeding on growth, maturation, feed utilization and flesh quality

A group of juvenile Atlantic halibut (mean, SD, 199.5 g, 44.7) was split into two experimental groups: one group was fed in excess twice a day and later five days a week (i.e. Control group), and the other group was starved for 5 weeks and then subsequently re-fed for 10 weeks (i.e. 5/10 starvation/re-fed group). This treatment was repeated for three years until the fish were harvested (mean final weight, 4.4 kg). A size +specific compensatory response was seen; partial compensation was observed in halibut weighing less than 500 g whereas full recovery growth was seen in halibut over 2 kg. Food consumption and feed conversion efficiency was measured during two full 5/10 starvation/re-fed cycles. The Control group consumed 86% more food in the measurement period, whereas the FCE 40% higher in the 5/10 starvation/re-fed group (FCE = 0.95) compared to the Control group (FCE = 0.68). In males lower gonadosomatic index (2.2 vs. 3.0) and plasma 11-ketotestosterone (0.22 vs. 1.26 ng ml^− 1) levels in the 5/10 starvation/re-fed group were observed during late winter and spring of 2006 (age 3 years) indicating higher age at 1st maturity. Starvation had a significant effect for improving the quality as starved fish had higher muscle pH (6.4 vs. 6.2), less gaping (0.1 vs. 0.3) and harder texture (61.4 vs. 57.6 N). Overall, the rearing on a repeated 5/10 starvation/re-fed regime for 3 years led to full growth compensation, higher feed conversion efficiency, lower male maturation and improved flesh quality.     

Validation of diagnostic assays to screen broodstock for Flavobacterium psychrophilum infections

It is hypothesized that the frequency of bacterial coldwater disease outbreaks can be reduced through the detection of the aetiologic agent, Flavobacterium psychrophilum, in broodstock followed by culling of eggs from heavily infected broodstock. Before a culling programme can be instituted, however, it is necessary to determine the sensitivity and specificity of existing assays for the detection of F. psychrophilum. In this study, tissue and ovarian fluid samples were collected from 224 fish at five hatcheries and screened using an enzyme‐linked immunosorbent assay (ELISA), a membrane‐filtration fluorescent antibody test (MF‐FAT), bacteriological culture and nested PCR. Latent class analysis was used to estimate sensitivity and specificity of kidney culture, kidney ELISA, nested PCR and MF‐FAT. Analytical sensitivity of the ELISA varied but was greatest when bacteria were cultured under iron‐limiting conditions. Diagnostic sensitivity estimates ranged from 0.02 (kidney culture) to 0.97 (kidney ELISA). Specificity estimates ranged from 0.02 (MF‐FAT) to 0.98 (kidney ELISA). In a separate challenge experiment, the ELISA confirmed the presence of F. psychrophilum in sub‐clinically infected fish. Results from this study demonstrate that the ELISA is an appropriate tool to screen broodstock and provides an indication of infection severity, which is crucial for implementation of a screening/culling programme.     

Investigations on the digestibility and metabolizability of the major nutrients and energy of maize leaves and barnyard grass in grass carp (Ctenopharyngodon idella)

In the uplands of northern Vietnam, culture of grass carp contributes significantly to income and household food security of Black Thai farmers. Maize is one of the most important upland crops and barnyard grass is the most important weed in the paddy rice fields. Thus, these are frequently used by small‐scale fish farmers as fish feed. An 8‐week feeding trial was conducted simultaneously in a recirculation and in a respirometric system to determine the digestibility and metabolizability of the nutrients of maize leaves and barnyard grass, to assess their crude protein, lipid and energy conversion and to estimate the energy allocation in grass carp. The following diets were used: diet ‘A’ (reference diet) containing 39% crude protein with 19.8 MJ kg^?1 gross energy; diets ‘B’, ‘C’ and ‘D’, which contained the same amount of the reference diet as the control diet, supplemented with a known amount of dried barnyard grass, dried maize leaves or fresh maize leaves, respectively. Five fish were assigned to each treatment in each experimental unit. Reference and test diets were fed to fish and faecal samples were collected and the oxygen consumption was measured in order to set up an energy budget of the fish over the whole experimental period. The weekly development of the body weight was recorded. The body weight gain of the fish fed diet D was significantly higher than that of the group fed diet A, which also in turn was significantly higher than that of fish fed diet B or C. The apparent digestibility coefficients (ADC) of nutrients and gross energy for the different experimental diets in fish kept in the aquaria were, for diets A, B, C and D respectively, 94.1%, 60.9%, 70.5% and 84.7% for protein, 91.3%, 60.7%, 76.8% and 71.8% for lipid; 95.9%, 44.5%, 60.6% and 69.1% for gross energy. The partial ADC of plant leaf ingredients were determined and barnyard grass and dried maize leaves were found to be not only poorly digestible but having negative impact on the digestibility of the reference diet, while fresh maize leaves were well digested. The results of the present study indicated that fresh maize leaves have a good potential to be used as supplement in diets for grass carp.     

Relative potencies of natural estrogens on vitellogenin and choriogenin levels in the Indian freshwater spotted snakehead, <Emphasis Type="Italic">Channa punctata</Emphasis>: in vivo and in vitro studies

The relative efficacies of three natural estrogens viz., estrone (E₁), estradiol-17β (E₂) and estriol (E₃) to induce synthesis of vitellogenin (Vg) and choriogenin (Chg) were assessed in primary hepatocyte cultures of the Indian freshwater spotted snakehead, Channa punctata. Hepatocytes were isolated from the spotted snakehead liver by a non-enzymatic protocol. Optimum culture conditions were standardized for ensuring their viability and functioning. Isolated hepatocytes were cultured for 48 h for monolayer formation and then exposed to various concentrations (0.001–10 μM) of the three estrogens. Competitive homologous ELISAs, developed and validated for spotted snakehead Vg and Chg were employed to determine the amounts of these two proteins secreted into the culture medium after 48 h of incubation. The results reveal that although all the three estrogens were effective in inducing the production of Vg and Chg in a dose-dependent manner, there were differences in their relative potencies. Of three estrogens, E₁ was the least potent and could induce synthesis of Vg and Chg only at a minimum concentration of 0.5 μM; whereas significant levels of both the proteins were quantified in culture medium by exposing the hepatocytes to E₂ or E₃ even at a concentration of 0.001 μM. All three estrogens were effective in inducing synthesis of Vg and Chg in vivo also. These results suggest the possibility of employing the above in vitro experimental design to monitor the presence of estrogens/estrogen-like chemicals in natural waters, which could interfere with the estrogen receptor system of fish. This study further points to the possibility of using Chg, in addition to Vg, as a parameter for screening various chemicals for their estrogenic activity.