Sub-lethal exposure to ultraviolet radiation reduces prey consumption by Atlantic cod larvae (Gadus morhua)

Levels of ultraviolet-B radiation (UV-B: 280–315 nm) reaching the earth’s surface have increased over the past few decades due to ozone depletion. It is well documented that exposure to UV-B radiation increases mortality in marine fish larvae. However, few studies have examined sub-lethal effects of UV-B radiation such as, for example, the possibility that it affects prey consumption by fish larvae. Atlantic cod larvae were exposed to a sub-lethal level of ultraviolet-B radiation (UV-B: 280–315 nm) for 15 h. After the exposure, rotifers (4/mL) were fed to cod larvae at three different post-exposure intervals (20 min, 3 and 6 h). Trials were replicated three times for each post-exposure interval. The number of rotifers in the gut and the percent of empty guts (number of fish with empty gut/number of fish examined × 100) were analyzed. Results were compared to those of unexposed fish (the control treatment). UV-B exposed cod larvae had consumed significantly fewer rotifers than control fish at all post-exposure intervals. There was no significant difference in the frequency of empty guts between fish in the UV-B treatment versus fish in the control treatment at any of the post-exposure time points (although the difference at 20 min post-exposure was borderline significant). These observations suggest that cod larvae exposed to sub-lethal levels of UV-B have lower net energy gain which may lead to reduced growth rate and possibly poorer survival.     

Foraging behavior in early Atlantic cod larvae (Gadus morhua) feeding on a protozoan (Balanionsp.) and a copepod nauplius (Pseudodiaptomussp.)

Food limitation is likely to be a source of mortality for fish larvae in the first few weeks after hatching. In the laboratory, we analyzed all aspects of foraging in cod larvae (Gadus morhua Linnaeus) from 5 to 20 d post-hatching using protozoa (Balanion sp.) and copepod nauplii (Pseudodiaptomus sp.) as prey. A camera acquisition system with two orthogonal cameras and a digital image analysis program was used to observe patterns of foraging. Digitization provided three-dimensional speeds, distances, and angles for each foraging event, and determined prey and fish larval head and tail positions. Larval cod swimming speeds, perception distances, angles, and volumes increased with larval fish size. Larval cod swam in a series of short intense bursts interspersed with slower gliding sequences. In 94% of all foraging events prey items were perceived during glides. Larval cod foraging has three possible outcomes: unsuccessful attacks, aborted attacks, and successful attacks. The percentage of successful attacks increased with fish size. In all larval fish size classes, successful attacks had smaller attack distances and faster attack speeds than unsuccessful attacks. Among prey items slowly swimming protozoans were the preferred food of first-feeding cod larvae; larger larvae had higher swimming speeds and captured larger, faster copepod nauplii. Protozoans may be an important prey item for first-feeding larvae providing essential resources for growth to a size at which copepod nauplii are captured. Received: 20 April 1999 / Accepted: 12 January 2000     

Role of bacteria with regard to chitin degradation in the digestive tract of the cod Gadus morhua

Numbers of aerobic, heterotrophic and chitinolytic bacteria in stomach, pyloric caeca and intestine of eight specimens of freshly caught cod (Gadus morhua L.) were determined by the spread plate method (medium: sea water agar, including 1% chitin). In crude enzyme extracts, chitinase activity was assayed according to the end product measurement. Results revealed no correlation between the number of chitinoclasts and the level of chitinase activity in any part of the digestive tract of cod. In another series of studies, antibiotics (Ampicillin/Sisomicin, or Chloramphenicol) were applied via tank water and feed. In most cases, antibiotic treatment resulted in the complete elimination of bacteria from the digestive tract. In contrast, up to 1.0×10⁹ chitinoclasts per g wet wt were determined in control fish. Chitinase did not reflect the absence or presence of bacteria: no significant difference in enzyme activities of cod treated with Ampicillin/Sisomicin and non-treated fish could be demonstrated. A decrease of chitinase activities of cod exposed to Chloramphenicol is interpreted as a toxic effect of this antibiotic on the fish. It is concluded that chitinase activities measured in the digestive tract of G. morhua primarily originate from fish tissues.Results presented in this paper are part of the author's Ph.D. dissertation. Authorization for pre-publication was given by the Graduation Committee of the Biology Department, University of Hamburg     

Feeding opportunities of larval and juvenile cod (Gadus morhua) in a Greenlandic fjord: temporal and spatial linkages between cod and their preferred prey

Feeding of fish depends on a spatial and temporal match with prey, and since larval and juvenile feeding can be highly selective, their preferences for given prey sizes and taxa should be considered when quantifying the actual availability of potential prey. We investigated the diet and prey preferences of the early-life stages of Atlantic cod (Gadus morhua) to quantify the availability of prey during a spring-summer season in a West Greenlandic fjord. We hypothesized that abundances of larval and juvenile cod at size were synchronized to optimal availability of preferred prey in space and time. The present analysis is based on nine cruises each covering 5 stations visited between 24 May and 5 August 2010 comparing zooplankton abundance, cod gut content and distribution patterns. Cod 4–25 mm in length preferred prey of about 5 % of their own length. During ontogeny, their preferences changed from calanoid nauplii towards Pseudocalanus spp. and Calanus spp. copepodites. The larvae/juvenile had an exceptionally high dietary contribution from cladocerans, which were highly preferred by cod larger than 9 mm, while the abundant Metridia longa and the non-calanoid copepods contributed less. These findings stress the importance of focusing on abundance of preferred prey when assessing the actual prey availability to young fish. We found a spatio-temporal overlap between cod and their preferred prey, and observations suggest that advection of both zooplankton and cod contributed to this overlap. Hence, the larval feeding opportunities might be sensitive to climate-related changes affecting the circulation patterns in this fjord.     

Distribution of enzymes of glycolysis and gluconeogenesis in fish tissues

Gluconeogenesis in fishes has been demonstrated in whole animals and liver preparations. However, at present, the relative physiological importance of possible substrates such as lactate, pyruvate and amino-acids or the precise sites of gluconeogenesis are unclear. In mammals, gluconeogenesis takes place in the liver and kidney, and the same could occur in fishes although it has been proposed that fish red muscle is also capable of reconverting lactate (derived from white muscle) to glucose. In this present study, the activities of 3 key glycolytic (hexokinase, phosphofructokinase and pyruvate kinase) and 2 key gluconeogenic (fructose diphosphatase and phosphoenolpyruvate carboxykinase) enzymes were investigated in tissues of the rainbow trout Salmo gairdneri, the cod Gadus morhua, and the plaice Pleuronectes platessa in order to elucidate the relative glycolytic/gluconeogenic capacities of the individual fish tissues. The glycolytic enzymes were found in all tissues, the relative potential being skeletal muscle>heart, brain >kidney, gills>liver. The gluconeogenic enzymes were not present in all tissues, and were mainly concentrated in the liver and kidney. Hence the results indicate that the liver, and to a lesser degree, the kidney are the major sites of gluconeogenesis in fishes, and that the process is unlikely to occur in skeletal muscle.     

Modeling the Effects of Fishing and Implications for the Design of Marine Protected Areas: Juvenile Fish Responses to Variations in Seafloor Habitat

Abstract: A number of recent studies have linked post-settlement survivorship of Atlantic cod (  Gadus morhua ) with the complexity of the seafloor to which fish settle. Survivorship is greater in habitats of higher complexity (e.g., pebble-cobble substratum with emergent epifauna> pebble-cobble> sand), where cover provides shelter from predators. Fishing with mobile gear such as bottom trawls and dredges reduces the complexity of seafloor habitats. We used a dynamic model to (1) link patterns in habitat-mediated survivorship of post-settlement juvenile cod with spatial variations in habitat complexity, (2) simulate habitat change based on fishing activities, and (3) determine the role of marine protected areas in enhancing recruitment success. Density-dependent natural mortality was specified as three alternative functional response curves to assess the influence of different predator foraging strategies on juvenile survivorship during the first 12 months of demersal existence. We applied the model to a theoretical patch of hard-bottom substrata and to a case study based on seafloor habitat distributions at Stellwagen Bank National Marine Sanctuary (Gulf of Maine, Northwest Atlantic). Our results demonstrate that patterns in the shape of response surfaces that show the relationship between juvenile cod survivorship and density as well as movement rate were similar regardless of functional response type, that juvenile cod movement rates and post-settlement density were critical for predicting the effects of marine protected-area size on survivorship, and that habitat change caused by fishing has significant negative effects on juvenile cod survivorship and use of marine protected areas can ameliorate such effects.     

Movements of farmed and wild Atlantic cod (Gadus morhua) released in Trinity Bay,Newfoundland

Fishery scientists and managers are investigating the feasibility of enhancing annual recruitment to the northern Atlantic cod (Gadus morhua L.) stock complex off Labrador and northeastern Newfoundland through the release of farmed fish back into the sea. Release of newly matured fish and adults with farm-advanced fecundity would increase the spawning biomass. Enhancement efforts might be measurably successful in major bays that are year-round habitats for cod. To determine if farmed cod would remain and spawn in Trinity Bay, 14 fish with surgically implanted transmitters were released in November 1992. Sonic tracking confirmed that farmed cod released on the western side of Trinity Bay overwintered within the bay, and integrated with wild cod approaching spawning condition in April 1993. Blood plasma antifreeze levels confirmed that these wild cod had overwintered inshore in subzero waters. A spawning aggregation was found in July 1993, providing evidence that northern cod reproduce in Newfoundland bays. These findings suggest that it may be possible to increase the number of cod spawning inshore through the release of farmed fish.     

Food and daily ration of the rock soleLepidopsetta bilineata (Pleuronectidae) in the eastern Bering Sea

A total of 129 specimens ofLepidopsetta bilineata (Ayres) were collected in the eastern Bering Sea (57°02–58°11N; 161°58–164°38W) between 28 August and 7 September 1984. Examination of its stomach and intestine contents revealed that polychaetes, gammarid amphipods and echiuroids constitute the bulk of its diet. Prey type is related to fish size. Juveniles consume more mobile prey, such as small crustaceans (gammarid amphipods, cumaceans, carideans), while adults, although still continuing to feed on gammarid amphipods, preferentially feed on infaunal organisms such as polychaetes, echiuroids and mollusks. Some prey, such as echinoderms, fishes and urochordates, were encountered only in stomachs, which resulted in a higher index of feeding diversity for this organ. Rock sole feeds primarily during daylight hours, peaking at dusk. Its daily ration was estimated as 0.49, 1.12, and 1.61% of fish body weight from contents of stomach, intestine, and stomach plus intestine, respectively.     

The hypoxia avoidance behaviour of juvenile Atlantic cod (<Emphasis Type="Italic">Gadus morhua</Emphasis> L.) depends on the provision and pressure level of an O<Subscript>2</Subscript> refuge

The frequency of low O₂ (hypoxia) has increased in coastal marine areas but how fish avoid deleterious water masses is not yet clear. To assess whether the presence and oxygen pressure (PO₂) level of an O₂ refuge affects the hypoxia avoidance behaviour of fish, individual Atlantic cod (Gadus morhua L.) were exposed to a range of O₂ choices in a 2-way choice chamber at 11.4°C over two different experiments. Cod in the first experiment were allowed access to a fixed O₂ refuge (fully air-saturated seawater) whilst oxygen pressure (PO₂) on the other side was reduced in steps to a critically low level, i.e. 4.3 kPa—a point where cod can no longer regulate O₂ consumption. Under these conditions, cod did not avoid any level of hypoxia and fish swimming speed also remained unchanged. In contrast, strong avoidance reactions were exhibited in a second experiment when fish were again exposed to 4.3 kPa but the safety, i.e. PO₂, of the refuge was reduced. Fish not only spent less time at 4.3 kPa as a result of fewer sampling visits but they also swam at considerably slower speeds. The presence of an avoidance response was thus strongly related to refuge PO₂ and it is unlikely that cod, and possibly other fish species, would enter low O₂ to feed in the wild if a sufficiently safe O₂ refuge was not available. It is therefore hypothesized that the feeding range of fish may be heavily compressed if hypoxia expands and intensifies in future years.     

Free amino acids as energy substrate in developing eggs and larvae of the cod Gadus morhua

The content of free amino acids (FAA) in the cod (Gadus morhua L.) egg is about 200 nmol at spawning, decreasing by about 100 nmol/egg during the egg stage and about 75 nmol/larva during the yolksac larval stage. Together, alanine, leucine, serine, isoleucine, lysine, and valine account for about 75% of the decrease. Ammonium accumulates gradually during the egg stage and is quickly excreted after hatching. The body protein content is maintained during the egg and yolksac larval stages. The measured oxygen uptake of the cod embryo during the egg and yolksac larval stages accounts for about 85% of the oxygen necessary to catabolize the FAA disappearing during this period. Ammonia excretion of the cod embryo, as taken from literature data, is similar to the expected ammonia production from catabolism of the FAA. Our data suggest that FAA are a major substrate for aerobic energy production in cod eggs and yolksac larvae. The implication of this finding for the production of a favourable first-feed for cod and other cultivated marine fish larvae, and for the selection of high quality eggs of marine fishes, is stressed.     

The foraging ecology of Arctic cod (Boreogadus saida) during open water (July–August) in Allen Bay, Arctic Canada

Arctic cod (Boreogadus saida) is a schooling fish providing a critical link between lower and upper trophic levels in the Arctic. This study examined foraging of Arctic cod collected from Allen Bay, Cornwallis Island, Canada (~75 N 95 W), during summer 2010 using temporal indicators of diet including stomach content, and carbon (δ¹³C) and nitrogen (δ¹⁵N) stable isotopes of liver and muscle. Foraging at the time of capture reflected sympagic and epi-benthic habitats indicated by the prevalence of cyclopoid and harpacticoid copepods in stomachs, whereas stable isotope data, which provide an estimate of feeding over a longer period, indicated pelagic prey as important. Prey selection of juveniles differed from adults based on stable isotopes, while large adults showed the most separation based on stomach contents, suggesting size-related diet shifts. Compared to studies near Resolute in the 1970s, 1980s, and 1990s, growth and pre-spawning gonadal conditions of Arctic cod have not changed.     

Escape speeds of marine fish larvae during early development and starvation

Response rates to tactile stimulation and subsequent escape speeds were measured using a video-recording system during early development and starvation of fish larvae. The species studied included the yolk-sac larvae of Clyde and Baltic herring (Clupea harengus L.), cod (Gadus morhua L.), flounder (Platichthys flesus L.) and older larvae of Clyde herring. The proportion of larvae responding (response rate) was initially about 20 to 25% in herring and 35 to 40% in cod and flounder using a probe, but about 70 to 80% using the sucking action of a pipette in all species except flounder. Both response rates and escape speeds (mean and maximum) tended to peak 1 to 2 d before the PNR (point-of-no-return, when 50% of larvae are too weak to feed), then decreased slowly during further starvation. An inter-species comparison showed that the highest recorded mean escape speeds (measured over a period of 200 ms) and highest maximum escape speeds (over 20 ms) ranged from 5.7 to 8.6 BL/s (body lengths/s) and 12.1 to 16.1 BL/s, respectively. The larvae made directional responses away from the stimulus only when they developed and reached the feeding stage.     

Trophic relationships of white-chinned petrels from Crozet Islands: combined stomach oil and conventional dietary analyses

The diet of white-chinned petrels Procellaria aequinoctialis breeding at the Crozet Archipelago (southern Indian Ocean) was studied using two complementary methods: lipid analysis of stomach oils as trophic markers together with the conventional dietary approach (i.e., stomach content analysis). Objectives were (1) to investigate the adult diet when they feed for themselves by analyzing stomach oil lipids, and (2) to compare the lipid signature of chick and adult oils. Stomach oils mainly consisted of triacylglycerols (TAG), diacylglycerol-ethers (DAGE) and wax esters (WE) (66, 14 and 11%, respectively). The dietary origin of TAG and WE was evaluated by linear discriminant analyses with fatty acid and fatty alcohol fractions. Analyses evidenced that stomach oils did not originate from Antarctic krill, but instead from myctophid fish, thus demonstrating the importance of mesopelagic fish in the nutrition of adult petrels. This result was consistent with the identification of digested remains of myctophids recovered from adult stomach contents after long foraging trips. Large amounts of a rare lipid class, DAGE (up to 76% of total lipids), were identified in two stomach oils, together with fresh remains of the squid Gonatus antarcticus (99% by mass), suggesting that DAGE could have the potential to be trophic markers of cephalopods. Moreover, six oils probably originated from Patagonian toothfish, thus confirming strong interactions between white-chinned petrels and fisheries. Comparison between chick and adult stomach oils indicated no major differences in their biochemical composition suggesting an identical dietary origin of oils, mainly myctophids. Both adult and chick oils can therefore be used to determine the feeding ecology of adult birds when they feed far away from their breeding grounds. Finally, food analysis of chick samples and adult samples collected after short and long trips indicated different foraging grounds during the two kinds of trips, and also between long trips performed in subtropical and Antarctic waters.     

Assessment of lead and cadmium residues in farmed fish in Machakos and Kiambu counties,Kenya

Lead (Pb) and cadmium (Cd) concentrations were determined in muscle, gonad, liver and brain of tilapia fish caught from fish ponds in Machakos and Kiambu counties in Kenya. A total of 217 fish samples were randomly sampled from the two counties. Acid digestion method and atomic absorption spectrophotometer were used for analysis. Heavy metal concentrations varied significantly depending on the type of tissue analyzed. Generally, the highest concentration of Pb was detected in brain and the liver. Fish organs contained Pb in the following order: brain > liver > muscle > gonad, while Cd followed the order: brain > liver > gonad > muscle. Kiambu county recorded higher concentration of the studied heavy metals compared to Machakos county although statistically there was no difference. Lead and Cd content in both counties studied exceeded the maximum allowable limit. The study recommended controlling industrial and agricultural effluents into surface water and proper sitting of ponds to minimize the risk of contamination of farmed fish by heavy metals.     

Pathological and toxicological investigations of chronic nitrate poisoning in cattle

In this study, three stillborn and five premature and blind calves from cows suffering from chronic nitrate poisoning were examined pathologically. The live calves were euthanized, due to a poor prognosis. Complete necropsy was performed on all the eight affected calves. Hydroperitoneum and ascites were the most predominant lesions in the stillborn calves. Edema and hemorrhage were observed in the lungs of all the calves. Hemorrhages were seen in the digestive systems of four calves. Blood samples were taken from five live calves and their mothers, and from two cows that had delivered stillborn or blind calves, for biochemical observations and measurement of methemoglobin levels. Blood values of five healthy cattle from a herd with no history of stillborn or blind calves were used as controls. Water and feed samples were taken from five different farms with a history of stillborn or blind calves and examined for nitrates. Nitrate and nitrite levels of feeds ranged from 154 to 480 and 1.5 to 20?ppm respectively; these levels can cause chronic poisoning. Levels of nitrates and nitrites in blood samples from cows and calves were found to be high. While hemoglobin values were found to be low, methemoglobin values were very high. No differences were observed between liver enzyme levels of the affected and control cattle. There were no bacteria or virus isolated from these animals, and histopathological examination revealed no indication of any other diseases.     

Changes in lipid content,lipid class composition and fatty acid composition of developing eggs and unfed larvae of cod (Gadus morhua)

Changes in the lipid class and fatty acid composition of developing eggs and unfed larvae of cod (Gadus morhua L.) were studied with the objective of determining probable requirements of cod larvae for dietary lipid. The eggs were collected on 24 March 1985 from holding tanks containing cod which had been caught off the northwest coast of Scotland. Phosphatidylcholine (PC) was the only lipid class to decline in absolute terms during embryogenesis. Catabolism of neutral lipid was initiated during the first week after hatching, and the rate of neutral lipid utilisation increased after the larvae had completely absorbed their yolk sacs. The quantity of triacylglycerol (TAG) remained constant during embryogenesis, but the percentage of 22:6(n-3) int TAG increased substantially during this period. It was calculated that ca. 33% of the 22:6(m-3) released during the process of PC catabolism was incorporated into TAG and sterol ester. The results suggest that PC, replete in appropriate essential fatty acids, should represent a major proportion of the lipid in artificial diets for fish and crustacean larvae.     

Fatty acid composition as an indicator of food intake in cod larvae Gadus morhua from Lofoten,Northern Norway

Fatty acid analyses were used to study the transition from endogenous to exogenous feeding in Arcto-Norwegian cod larvae Gadus morhua L. sampled from Lofoten waters, Northern Norway in April 1985. Fatty acids of total lipids were analysed from phytoplankton, eggs and nauplii of Calanus finmarchicus, and cod eggs and larvae. Gas chromatographic and gas chromatographic-mass spectrometric methods were used. On the basis of these analyses it is suggested that lipids of phytoplankton origin form an important part of the diet of cod larvae during the first feeding period.     

Matching and mismatching stable isotope (δ13C and δ15N) ratios in fin and muscle tissue among fish species: a critical review

Using non-lethal tissue sampling for stable isotope analysis has become standard in many fields, but not for fishes, despite being desirable when species are rare or protected, when repeated sampling of individuals is required or where removal may bias other analyses. Here, we examine the utility of fish dorsal fin membrane as an alternative to muscle for analyzing δ¹³C and δ¹⁵N ratios in two reef fish species (blue cod Parapercis colias and spotty Notolabrus celidotus) that have differing feeding modes. Both species exhibited evidence of size-based feeding from fin δ¹⁵N values, but not from muscle. Blue cod fin δ¹⁵N increased steadily throughout the sampled size range (213–412 mm fork length), whereas spotty exhibited a distinct ontogenetic diet shift at approximately 120–140 mm fork length after which size-based feeding did not occur. Fin membrane was higher than muscle in δ¹³C in both species and in δ¹⁵N for blue cod, but fin δ¹⁵N was lower than muscle in spotty. The δ¹³C and δ¹⁵N fin–muscle offsets were constant in spotty regardless of size, while in blue cod, δ¹³C was constant with fish size, but δ¹⁵N offsets increased with increasing fish size. Non-lethal sampling utilizing fin tissue can be employed to estimate stable isotope values of muscle in fishes, but it is necessary to assess relationships among tissues and the effects of fish size on isotope values a priori for each species studied. Our data indicated that fin membrane may be a more sensitive tissue than muscle for detecting size-based feeding in some fish species using stable isotopes. A critical literature review revealed inconsistencies in tissue types tested, little understanding of tissue-specific trophic shift or turnover rates, and pseudo-replicated analyses leading to erroneous postulating of 1:1 relationships between tissues.     

Element levels in cultured and wild sea bass (Dicentrarchus labrax) and gilthead sea bream (Sparus aurata) from the Adriatic Sea and potential risk assessment

In this study, the role of aquaculture activity as a source of selected metals was analyzed. Significant differences in element content between cultured (Dicentrarchus labrax, Sparus aurata) and wild fishes as well as between fish muscle and their feed were detected. Higher concentrations of trace elements (i.e., As, Cu, Hg, Se) in wild fish tissues in comparison with cultured ones indicate additional sources of metals beside fish feed as natural and/or anthropogenic sources. Generally, mean Cd, Cu, Pb, Se, and Zn concentrations in cultured (0.016, 1.79, 0.14, 0.87, and 34.32 μg/g, respectively) and wild (0.011, 1.97, 0.10, 1.78, and 23,54 μg/g, respectively) fish samples were below the permissible levels, while mean As (2.57 μg/g in cultured, 4.77 μg/g in wild) and Cr (5.25 μg/g in cultured, 2.92 μg/g in wild) values exceeded those limits. Hg values were lower in cultured (0.17 μg/g) and higher in wild (1.04 μg/g) fish specimens. The highest elemental concentrations were observed in almost all fish samples from Kor?ula sampling site. The smallest cultured sea basses showed As (4.01 μg/g), Cr (49.10 μg/g), Pb (0.65 μg/g), and Zn (136 μg/g) concentrations above the recommended limits; however, values decreased as fish size increased. Therefore, the majority of metal concentrations in commercial fishes showed no problems for human consumption. Also calculated Se:Hg molar ratios (all >1) and selenium health benefit values (Se-HBVs) (all positive) showed that consumption of all observed fishes in human nutrition is not risk.     

Liver Metallothionein Level and Metal Content in Fish of Chung-Kung Stream,Taiwan-Tilapia and Liza Macrolepsis

A survey of Chung-Kung sttream fish was undertaken on six occasions from March 1994 to August 1995. Tilapia (Oreochromis hybrid) and Liza macrolepsis were the major species found downstream and were chosen as the target species for metallothionein (MT) analysis of liver tissue. in total MT of 77 tilapia and 48 Liza macrolepsis liver samples were analyzed. Significantly higher values (p >0.05) were found for the August 1995 samples than for samples taken earlier at site B (6 km from the sea) for tilapia and at site A (estuary) for Liza macrolepsis. Liver zinc, copper and cadmium were analyzed for the 1995 samples. for tilapia, liver MT concentration was found to correlate well with zinc concentration (r = 0.84, p > 0.001). for Liza macrolepsis, good correlation was found for MT with both zinc (r = 0.89, p > 0.001) and copper (r = 0.90, p > 0.001). These results indicate that in the Chung-Kung stream, zinc and copper may be the major inducers of fish liver MT. Since these two species are ubiquitous in tropical areas, they would provide a good biomarker for evaluation of integrated metal exposure in the tropical aquatic environment.