Complex physiological traits, such as routine aerobic metabolic rate or exercise performance, are indicators of the functional integrity of fish that can reveal sub-lethal toxicological effects of aquatic pollutants. These traits have proved valuable in laboratory investigations of the sub-lethal effects of heavy metals, ammonia and various xenobiotics. It is not known, however, whether they can also function as biomarkers of the complex potential range of effects upon overall functional integrity caused by exposure to mixtures of chemicals in polluted natural environments. The current study used portable swimming respirometers to compare exercise performance and respiratory metabolism of fish exposed in cages for three weeks to either clean or polluted sites on three urban European river systems: the river Lambro, Milan, Italy; the rivers Blythe, Cole and Tame, Birmingham, UK; and the river Amstel, Amsterdam, The Netherlands. The UK and Italian rivers were variously polluted with high levels of both bioavailable heavy metals and organics, and the Amstel by mixtures of bioavailable organics at high concentrations. In both the UK and Italy, indigenous chub (Leuciscus cephalus) exposed to clean or polluted sites swam equally well in an initial performance test, but the chub from polluted sites could not repeat this performance after a brief recovery interval. These animals were unable to raise the metabolic rate and allocate oxygen towards exercise in the second trial, an effect confirmed in successive campaigns in Italy. Swimming performance was therefore a biomarker indicator of pollutant exposure in chub exposed at these sites. Exposure to polluted sites on the river Amstel did not affect the repeat swimming performance of cultured cloned carp (Cyprinus carpio), indicating either a species-specific tolerance or relative absence of heavy metals. However, measurements of oxygen uptake during swimming revealed increased rates of routine aerobic metabolism in both chub and carp at polluted sites in all of the rivers studied, indicating a sub-lethal metabolic loading effect. Therefore, the physiological traits of exercise performance and metabolic rate have potential as biomarkers of the overall sub-lethal toxic effects of exposure to complex mixtures of pollutants in rivers, and may also provide insight into why fish do not colonize some polluted environments.

Background. Great cormorants (Phalacrocorax carbo L.) show the highest known foraging yield for a marine predator and they are often perceived to be in conflict with human economic interests. They are generally regarded as visually-guided, pursuit-dive foragers, so it would be expected that cormorants have excellent vision much like aerial predators, such as hawks which detect and pursue prey from a distance. Indeed cormorant eyes appear to show some specific adaptations to the amphibious life style. They are reported to have a highly pliable lens and powerful intraocular muscles which are thought to accommodate for the loss of corneal refractive power that accompanies immersion and ensures a well focussed image on the retina. However, nothing is known of the visual performance of these birds and how this might influence their prey capture technique. Methodology/ Principal Findings. We measured the aquatic visual acuity of great cormorants under a range of viewing conditions (illuminance, target contrast, viewing distance) and found it to be unexpectedly poor. Cormorant visual acuity under a range of viewing conditions is in fact comparable to unaided humans under water, and very inferior to that of aerial predators. We present a prey detectability model based upon the known acuity of cormorants at different illuminances, target contrasts and viewing distances. This shows that cormorants are able to detect individual prey only at close range (less than 1 m). Conclusions/Significance. We conclude that cormorants are not the aquatic equivalent of hawks. Their efficient hunting involves the use of specialised foraging techniques which employ brief short-distance pursuit and/or rapid neck extension to capture prey that is visually detected or flushed only at short range. This technique appears to be driven proximately by the cormorant's limited visual capacities, and is analogous to the foraging techniques employed by herons.

DNA strand breaks [as determined by the conventional and formamidopyrimidine glycosylase (FPG)-modified Comet assay] and antioxidant defense status [as indicated by superoxide dismutase (SOD) activity and reduced glutathione (GSH) concentration] were evaluated in healthy adult chub ( Leuciscus cephalus ) after exhaustive exercise [swimming to their critical swimming speed ( U crit ), twice in succession with a 40 min rest period between] vs. confined (unexercised) control fish. The conventional Comet assay revealed significantly higher DNA strand breaks in all the tissues (blood, liver, and gill), with the highest increase over background evident in the epithelial gill cells of swum fish compared to the controls. Moreover, when the FPG-modified Comet assay was conducted to reveal specific oxidative lesions, the gill cells of exercised fish sustained the highest level of oxidative DNA damage in comparison to the control. Data on tissue antioxidant defense mechanism were less conclusive, with no significant differences in the tissue levels of SOD or GSH. This suggests that either the degree of oxidative stress was not great enough to evoke a response in terms of defense mechanisms or the timescale of antioxidant defense response was somewhat different from the time between the application of stress and subsequent tissue sampling. From the swimming data, U crit was significantly lower on the second assessment compared to the first (repeat ratio: 0.76), suggesting that the fish were exercised to a level which was not sustainable. Overall, these findings support the theory that acute extreme exercise could result in oxidative stress and associated DNA damage in fish. These observations suggest that fish living in fast flowing and polluted rivers are at increased risk of DNA damage. Environ. Mol. Mutagen., 2006. © 2006 Wiley-Liss, Inc.

DNA strand breaks [as determined by the conventional and formamidopyrimidine glycosylase (FPG)‐modified Comet assay] and antioxidant defense status [as indicated by superoxide dismutase (SOD) activity and reduced glutathione (GSH) concentration] were evaluated in healthy adult chub (Leuciscus cephalus) after exhaustive exercise [swimming to their critical swimming speed (Ucrit), twice in succession with a 40 min rest period between] vs. confined (unexercised) control fish. The conventional Comet assay revealed significantly higher DNA strand breaks in all the tissues (blood, liver, and gill), with the highest increase over background evident in the epithelial gill cells of swum fish compared to the controls. Moreover, when the FPG‐modified Comet assay was conducted to reveal specific oxidative lesions, the gill cells of exercised fish sustained the highest level of oxidative DNA damage in comparison to the control. Data on tissue antioxidant defense mechanism were less conclusive, with no significant differences in the tissue levels of SOD or GSH. This suggests that either the degree of oxidative stress was not great enough to evoke a response in terms of defense mechanisms or the timescale of antioxidant defense response was somewhat different from the time between the application of stress and subsequent tissue sampling. From the swimming data, Ucrit was significantly lower on the second assessment compared to the first (repeat ratio: 0.76), suggesting that the fish were exercised to a level which was not sustainable. Overall, these findings support the theory that acute extreme exercise could result in oxidative stress and associated DNA damage in fish. These observations suggest that fish living in fast flowing and polluted rivers are at increased risk of DNA damage. Environ. Mol. Mutagen., 2006. © 2006 Wiley‐Liss, Inc.

SUMMARY Adult brown trout (Salmo trutta) were acclimatised to and maintained at seasonal temperatures (5°C in winter; 15°C in summer)and acclimated to reversed seasonal temperatures (15°C in winter; 5°C in summer) while exposed to the natural (i.e. seasonally variable)photoperiod. The mean critical swimming speeds (Ucrit) of animals acclimatised to the seasonal temperatures were similar, but more than 30% greater than those for fish acclimated to the reversed seasonal temperatures. The lower values of Ucrit that accompanied acclimation to reversed seasonal temperatures appeared largely to result from the inability of white muscle to function maximally, since the concentrations of lactate and ammonia in white muscle of fish swum to Ucrit at reversed seasonal temperatures were significantly lower than those in fish swum at seasonal temperatures. These observations,together with biochemical and morphometric attributes of muscle tissue,suggest that swimming ability is influenced, at least in part, by seasonal factors other than temperature. These data have important implications for the design of experiments using fish that experience predictable, usually seasonal, changes in their natural environment (temperature, dissolved oxygen,changes in water levels, etc.).

This study forms part of an investigation into the effects on fish of immersion in three rivers around Birmingham, UK. The rivers Blythe, Cole and Tame exhibit relatively high, intermediate and poor overall water quality, respectively, according to combined levels of polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs), as well as heavy metals. Specifically, biomarkers of genotoxicity (DNA strand breaks and adducts) were measured in feral and caged chub (Leuciscus cephalus), complementing another study in which data were presented for a number of other hepatic biomarkers measured in the same animals. In both feral and caged chub, there was a general elevation of DNA strand breaks with a decrease in chemical water quality, with some time points exhibiting significantly higher levels at the most (Tame) compared with least polluted sites (Blythe), particularly in the cage-held animals. Combined-season DNA adduct data suggested a higher degree of toxic insult in the feral compared with caged chub and revealed particularly high levels of adducts in fish caught from the Cole. The pattern of adducts shown was typical of exposure to a complex mixture of PAHs which were relatively high, and similar, in both the Cole and Tame. Overall, these data are consistent with exposure of both feral and caged chub to contaminants which are able to induce specific, moderately genotoxic effects.

ABSTRACT Adult brown trout were acclimated for 2–4 weeks to artificial soft water ([Ca2+] 25 μmol l−1) at neutral pH and at summer (15°C) temperature. During this period they swam against a current of approximately 0.25 ms −1. They then had their dorsal aorta cannulated and were exposed to neutral or sublethal pH (4.5) for 4 days in still water. After 4 days of exposure to sublethal pH, critical swimming speed (Ucrit) was 35% lower than that for fish at neutral pH. There were significant increases in arterial and in blood lactate concentrations at Ucrit compared with the values in resting fish at neutral pH and these led to significant reductions in plasma pH. There were no such changes in fish at sublethal pH. There were no significant changes in intracellular pH (pHi) of red blood cells at Ucrit, probably as a result of increases in the levels of plasma catecholamines. There were significant reductions in pHi of red and white muscle fibres at Ucrit. It is argued that these values were not as low in the white fibres as those seen in previous studies after fish have been chased to exhaustion and, therefore, that the fish in the present study were not completely exhausted, although they would no longer swim at a steady speed. As pHi of the red muscle was the same at Ucrit for fish at neutral and at sublethal pH, it is suggested that Ucrit (fatigue) coincides with a particular pHi of the red muscles and possible mechanisms are discussed.