Pesticide-contaminated feeds in integrated grass carp aquaculture: toxicology and bioaccumulation.

Effects of dissolved pesticides on fish are widely described, but little is known about effects of pesticide-contaminated feeds taken up orally by fish. In integrated farms, pesticides used on crops may affect grass carp that feed on plants from these fields. In northern Vietnam, grass carp suffer seasonal mass mortalities which may be caused by pesticide-contaminated plants. To test effects of pesticide-contaminated feeds on health and bioaccumulation in grass carp, a net-cage trial was conducted with 5 differently contaminated grasses. Grass was spiked with 2 levels of trichlorfon/fenitrothion and fenobucarb. Unspiked grass was used as a control. Fish were fed at a daily rate of 20% of body mass for 10 d. The concentrations of fenitrothion and fenobucarb in pond water increased over time. Effects on fish mortality were not found. Fenobucarb in feed showed the strongest effects on fish by lowering feed uptake, deforming the liver, increasing blood glucose and reducing cholinesterase activity in blood serum, depending on feed uptake. Fenobucarb showed increased levels in flesh in all treatments, suggesting bio-concentration. Trichlorfon and fenitrothion did not significantly affect feed uptake but showed concentration-dependent reduction of cholinesterase activity and liver changes. Fenitrothion showed bioaccumulation in flesh which was dependant on feed uptake, whereas trichlorfon was only detected in very low concentrations in all treatments. Pesticide levels were all detected below the maximum residue levels in food. The pesticide-contaminated feeds tested did not cause mortality in grass carp but were associated with negative physiological responses and may increase susceptibility to diseases.

Influence of dietary non-essential amino acid profile on growth performance and amino acid metabolism of Nile tilapia, Oreochromis niloticus (L.).

The quality of dietary protein is an important factor influencing the growth performance of fish. To evaluate the quality of protein, the variables commonly studied are the composition of the essential amino acids, the digestibility and the protein use efficiency. The goal of the present experiment was to test the effect of the dietary non-essential amino acid composition on the growth of Nile tilapia (Oreochromis niloticus). The fish were fed three purified diets differing only in their non-essential amino acid composition. The influence of the experimental diets on the growth performance, on the activity of enzymes involved in the amino acid metabolism, aspartate aminotransferase (ASAT) and alanine aminotransferase (ALAT), and on whole body delta(15)N values was investigated. Body mass, lipid, protein and energy gain differed significantly between the feeding groups. The activity of ASAT in the whole liver was significantly higher in fish with a positive protein balance compared to fish which lost protein. Whole body delta(15)N values of fish were negatively correlated with their body mass gain. Despite the poor utilisation of synthetic amino acids, the experiment indicates the importance of the dietary non-essential amino acid composition for the growth performance of fish. The study reveals the possibility to trace the utilisation of synthetic amino acids by determining the isotopic composition of dietary amino acids and tissues or whole bodies of animals.

Improving estimates of trophic shift in Nile tilapia, Oreochromis niloticus (L.), using measurements of lipogenic enzyme activities in the liver.

To test whether the measurement of selected enzyme activities could be used to estimate more precisely the trophic shift of C isotopes, Nile tilapia (Oreochromis niloticus) were fed semi-synthetic diets differing in their lipid contents (1.7%, 5.0%, 10.8% and 20.0%). The diets were formulated to contain the same amount of nitrogen and metabolizable energy and were made from casein, wheat starch, corn germ oil supplemented with vitamins, minerals and L-arginine. The influence of the different diets on the activity of two lipogenic enzymes, ATP-citrate lyase and malic enzyme, on delta13C values in the whole fish, the liver and their correlation was investigated. There was a strong positive correlation between delta13C values in the lipids of whole fish and those of their livers. The activities of lipogenic enzymes increased significantly with increasing trophic shift of C isotopes (Deltadelta13Cdiet-fish values) in the lipids. If the relationship between trophic shift and enzyme activity can be confirmed in situations where feed quantity and quality are not known, the determination of enzyme activities would enable better estimates of the trophic shift to be made thus significantly improving back-calculation of diets from stable isotope data.

Feeding level and diet quality influence trophic shift of C and N isotopes in Nile tilapia (Oreochromis niloticus (L.)).

Many scientists use naturally occurring stable isotopes to reconstruct the diets of animals. However, isotopic ratios may be affected not only by the composition of the diet but also by the amount of food consumed. Thus, an experiment using tilapia (Oreochromis niloticus) was carried out to test the influence of feeding level on delta13C and delta15N of fish given a semi-synthetic wheat gluten/wheat starch based diet. In addition, the effect of diet quality was tested by comparing tilapia given this feed with tilapia fed a fish meal/wheat meal based diet. Forty-four tilapia were reared individually. After a prefeeding phase, fish were randomly assigned to five groups, four on the semi-synthetic diet at different feeding levels and one group on the fish meal/wheat meal based diet fed at the equivalent of the highest level of the semi-synthetic diet. The experiment lasted eight weeks. Proximate composition, gross energy content and delta13C and delta15N values were determined in feed and fish, for delta13C separately in the lipids and the lipid-free matter. Delta13C in the lipids and the lipid-free matter and delta15N of tilapia fed the semi-synthetic diet decreased significantly with increasing feeding rate. The absolute values of the trophic shift in fish fed the semi-synthetic wheat based diet were significantly higher than in fish fed the fish meal/wheat meal based diet. The different delta13C and delta15N values in tilapia fed the same diet at different feeding levels and the influence of feed quality on the trophic shift add to the uncertainty involved in the use of stable isotopes in ecological research.

Effect of different lipid extraction methods on delta13C of lipid and lipid-free fractions of fish and different fish feeds.

For many ecological applications of stable carbon isotope techniques, it is necessary to separate the lipid and lipid-free fractions. The effect of different lipid extraction methods on the isotope signature of the remaining lipid-free matter as well as the lipid fraction was tested. A hot extraction form of the Soxhlet method using petrol-ether was compared with two liquid-liquid extraction methods for lipid determination described by Bligh and Dyer and Smedes. Solid samples of fish and different natural food items were subjected to extraction and the carbon isotope ratios in lipid and lipid-free matter determined by IRMS. All methods were suitable for lipid extraction from all samples analysed here and did not cause biologically relevant differences (> 1%) in carbon isotopic ratios, except the Bligh and Dyer extraction method using chloroform which caused systematic errors for delta13C when applied to diatoms.

Development of a low-cost technology for mass production of the free-living nematode Panagrellus redivivus as an alternative live food for first feeding fish larvae.

The free-living nematode Panagrellus redivivus is a suitable food source for first feeding fish. In the present report, a new method for the mass production of P. redivivus is presented. The technique involves multiplication of the nematode in monoxenic (single microorganism: Saccharomyces cerevisiae) solid culture (fluid media supported by 1- to 4-cm(3) sponge cubes) in autoclavable plastic bags (size range: 50 x 30 cm to 75 x 67 cm). Two growing media were tested: oat-meal medium (OM), which is an oat-based medium (16.7% oat-meal flour in 0.8% saline solution), and purified ingredient medium (PIM), a semi-synthetic medium (1.64% meat peptone, 0.94% yeast extract, 12.6% corn starch, 0.24% glucose, 1.48% sunflower oil, in 0.8% saline solution). The bags were inoculated with 350 nematodes/g medium. After an average period of 12 days (11-13 days) at 25 degrees C, the average yield (number of nematodes/g medium) was 241 x 10(3) for OM and 333 x 10(3) for PIM in 12-l bags (50 x 30 cm). The production scale has currently reached a bag volume of 50 l (75 x 67 cm); using PIM and the conditions described above, it was possible to harvest more than 1.3 x 10(9) nematodes/bag (291 x 10(3) nematodes/g medium). In PIM, when sun flower oil was replaced with the same amount of fish oil or cod liver oil, yields of 259 x 10(3) and 290 x 10(3) nematodes/g medium, respectively, were attained. The technology for mass production and formulation of P. redivivus should enable fish-hatchery operators to rely on a cheap, standardised, and permanently available live food product for first feeding fish larvae.

Stable isotopes in animal ecology: the effect of ration size on the trophic shift of C and N isotopes between feed and carcass.

The feeding ecology of living or extinct animal species is now frequently studied by analysis of stable isotope ratios in small quantities of carcass or remains such as bones or teeth. Although there are numerous papers on these applications in natural systems, the theoretical and experimental basis of this method is weak. In order to evaluate the effect of different feeding levels on the carbon and nitrogen trophic shift, an experiment was carried out in which fish (Nile tilapia, Oreochromis niloticus, initial weight 40.8 g) were fed for 4 weeks at three levels from slightly above maintenance to almost satiation. For each treatment, three fish were reared individually. The isotopic ratios of carbon and nitrogen in feeds and fish carcasses were determined and in the case of carbon this was done separately for lipids and lipid-free matter. The trophic shift was calculated at each feeding level from the delta13C and delta15N ratios of feed and fish. There was a significant trend towards higher values for the trophic shift at higher feeding rates in all fractions analysed. Although further research is required, it can be concluded that the effect of feeding level cannot be ignored when the diet of an animal has to be calculated from the ratios of isotopes in its body.

Metabolic fractionation of stable carbon isotopes: implications of different proximate compositions for studies of the aquatic food webs using δ13C data.

In two laboratory experiments using tilapia (Oreochromis niloticus) and common carp (Cyprinus carpio), we investigated the effect of lipid content in the fish carcass on the stable carbon isotope ratio (δ13C). In both experiments, a significant negative correlation was found between lipid content in the carcass dry matter and δ13C of total dry matter, but there was no influence on the δ13C of fat-free dry matter or lipids. As the lipid content of fish is known to vary with reproductive stage, season or nutritional state, separate analysis of fractions of the proximate composition of dry matter will lead to more reliable results than analysing the whole body. The differences in δ13C between diet and fish carcass (trophic shift) were different for the two species, calling for feeding trials under controlled conditions using the species and the feeds in question before applying the stable isotope tracer technique in the analysis of food webs.