No abstract

Alterations in the antioxidant cellular system have often been proposed as biomarkers of pollutant-mediated toxicity. This study evaluated the effects of mercury on oxidative stress biomarkers and bioaccumulation in the liver, gills, white muscle and heart of the freshwater fish matrinxã, Brycon amazonicus, exposed to a nominal and sub-lethal concentration (~20% of 96 h-LC(50)) of 0.15 mg L(-1) of mercury chloride (HgCl(2)) for 96 h in a static system. Increases in superoxide dismutase, catalase, glutathione peroxidase (GPx), glutathione S-transferase (GST) and glutathione reductase (GR) were observed in all tissues after HgCl(2) exposure, except for white muscle GR activity and hepatic GPx. In the liver and gills, the exposure to HgCl(2) also induced significant increases in reduced glutathione (GSH). Conversely, exposure to HgCl(2) caused a significant decrease in the GSH levels and an increase in the oxidized glutathione (GSSG) content in the white muscle, while both GSH and GSSG levels increased significantly in the heart muscle. Metallothionein concentrations were significantly high after HgCl(2) exposure in the liver, gills and heart, but remained at control values in the white muscle. HgCl(2) exposure induced oxidative damage, increasing the lipid peroxidation and protein carbonyl content in all tissues. Mercury accumulated significantly in all the fish tissue. The pattern of accumulation follows the order gills > liver >> heart > white muscle. In conclusion, these data suggest that oxidative stress in response to inorganic mercury exposure could be the main pathway of toxicity induced by this metal in fish.

Predominantly, Hoplias malabaricus inhabits stagnant 0, poor environments, whereas Hoplias lacrrdae occurs in well-aerated streams. The present study evaluates the influence of mode of life on 0, uptake and gill ventilation in equally-sized (300 g) specimens of this genus at 25-C. Comparing the species, H . lacerdae was characterized by the highest 0, uptake and gill ventilation combined with a relatively higher cost of breathing and a lower 0, extraction. Both species substantially increased ventilation in response to hypoxia with the difference that H. malaharicus exclusively augmented tidal volume, whereas H. lacerdae also increased breathing frequency.

The African sharptooth catfish Clarias gariepinus has bimodal respiration, it has a suprabranchial air-breathing organ alongside substantial gills. We used automated bimodal respirometry to reveal that undisturbed juvenile catfish (N=29) breathed air continuously in normoxia, with a marked diurnal cycle. Air breathing and routine metabolic rate (RMR) increased in darkness when, in the wild, this nocturnal predator forages. Aquatic hypoxia (20% air saturation) greatly increased overall reliance on air breathing. We investigated whether two measures of risk taking to breathe air, namely absolute rates of aerial O 2 uptake (Ṁ O2,air ) and the percentage of RMR obtained from air (%Ṁ O2,air ), were influenced by individual standard metabolic rate (SMR) and boldness. In particular, whether any influence varied with resource availability (normoxia versus hypoxia) or relative fear of predation (day versus night). Individual SMR, derived from respirometry, had an overall positive influence on Ṁ O2,air across all contexts but a positive influence on %Ṁ O2,air only in hypoxia. Thus, a pervasive effect of SMR on air breathing became most acute in hypoxia, when individuals with higher O 2 demand took proportionally more risks. Boldness was estimated as time required to resume air breathing after a fearful stimulus in daylight normoxia (T res ). Although T res had no overall influence on Ṁ O2,air or %Ṁ O2,air , there was a negative relationship between T res and %Ṁ O2,air in daylight, in normoxia and hypoxia. There were two T res response groups, 'bold' phenotypes with T res below 75 min (N=13) which, in daylight, breathed proportionally more air than 'shy' phenotypes with T res above 115 min (N=16). Therefore, individual boldness influenced air breathing when fear of predation was high. Thus, individual energy demand and personality did not have parallel influences on the emergent tendency to take risks to obtain a resource; their influences varied in strength with context.

To determine the location and distribution of chemoreceptors involved in the cardiovascular and respiratory responses to hypoxia of traira (Hoplias malabaricus), we measured heart rate, arterial blood pressure, ventilation frequency and amplitude of opercular movements during exposure to hypoxia and application of NaCN to either water bathing the gills (external) or the ventral aortic blood (internal). This was done before and after selective denervation of branchial branches of the IXth and Xth cranial nerves to various gill arches. The data suggest that hypoxia elicits a bradycardia that arises from internal receptors located in the first gill arch. They also indicate the presence of branchial and extra branchial O2-chemoreceptors that reflexively elevate systemic vascular resistance during hypoxia. Hypoxia induced increases in ventilation frequency arose primarily from external receptors located exclusively within the gills while increases in breathing amplitude also involved extra branchial receptors. In addition, the data suggest there are O2 sensitive chemoreceptors located in the first gill arch that attenuate the respiratory responses.

Oxidative stress biomarkers, in vivo heart rate (f (H)), and contraction dynamics of ventricle strips of bullfrog (Lithobates catesbeiana) tadpoles were evaluated after 48 h of exposure to a sub-lethal concentration (1 ppm) of the herbicide Roundup Original (glyphosate 41%). The activities of the antioxidant enzymes superoxide dismutase and catalase were increased in the liver and decreased in muscle, while oxidative damage to lipids increased above control values in both tissues, showing that the generation of reactive oxygen species and oxidative stress are involved in the toxicity induced by Roundup. Additionally, tadpoles' hyperactivity was associated with tachycardia in vivo, probably due to a stress-induced adrenergic stimulation. Ventricle strips of Roundup-exposed tadpoles (R-group) presented a faster relaxation and also a higher cardiac pumping capacity at the in vivo contraction frequency, indicating that bullfrog tadpoles were able to perform cardiac mechanistic adjustments to face Roundup-exposure. However, the lower maximal in vitro contraction frequency of the R-group could limit its in vivo cardiac performance, when the adrenergic-stimulation is present. The association between the high energetic cost to counteract the harmful effects of this herbicide and the induction of oxidative stress suggest that low and realistic concentrations of Roundup can have an impact on tadpoles' performance and success, jeopardizing their survival and/or population establishment.

The oxygen uptake ( i'Ol). breathing frequency (.f,), breath volume (V,,), gill ventilation ( bG) and oxygen extraction ( O h ) from the ventilatory current of four groups of 0rrochronii.s tiilvricirs during graded hypoxia were measured under the following acclimation temperatures: 20. 25. 30 and 35 C. The critical oxygen tensions (P,Oz). determined from VO, v. PO, of inspired water at each experimental temperature were. respectively. 19. I f 3.1. 18.0k4.9. 29.7 1 4 . I and 30.2 t 0.6 m m H g The.lk remained nearly constant during the reductions of 0, at all the temperatures studied. b' G increased discretely from normoxic levels until the P,O, was reached. below which it assumed extremely high values (17-fold higher or more). The increases observed in C' G resulted. at all the acclimation temperatures, in an elevation in V, rather than in,f,. The extraction of O1 dccreased gradually from normoxia until the P,O1 was reached. below which an abrupt reduction ofewtraction was recorded. except at 35 C when fish showcd a gradual reduction in extraction just below the tension of 80 mmHg.

SUMMARY Autonomic control of heart rate variability and the central location of vagal preganglionic neurones (VPN) were examined in the rattlesnake(Crotalus durissus terrificus), in order to determine whether respiratory sinus arrhythmia (RSA) occurred in a similar manner to that described for mammals. Resting ECG signals were recorded in undisturbed snakes using miniature datalogging devices, and the presence of oscillations in heart rate (fh) was assessed by power spectral analysis (PSA). This mathematical technique provides a graphical output that enables the estimation of cardiac autonomic control by measuring periodic changes in the heart beat interval. At fh above 19 min-1spectra were mainly characterised by low frequency components, reflecting mainly adrenergic tonus on the heart. By contrast, at fhbelow 19 min-1 spectra typically contained high frequency components, demonstrated to be cholinergic in origin. Snakes with a fh >19 min-1 may therefore have insufficient cholinergic tonus and/or too high an adrenergic tonus acting upon the heart for respiratory sinus arrhythmia (RSA) to develop. A parallel study monitored fh simultaneously with the intraperitoneal pressures associated with lung inflation. Snakes with a fh<19 min-1 exhibited a high frequency (HF) peak in the power spectrum,which correlated with ventilation rate (fv). Adrenergic blockade by propranolol infusion increased the variability of the ventilation cycle, and the oscillatory component of the fh spectrum broadened accordingly. Infusion of atropine to effect cholinergic blockade abolished this HF component, confirming a role for vagal control of the heart in matching fh and fv in the rattlesnake. A neuroanatomical study of the brainstem revealed two locations for vagal preganglionic neurones (VPN). This is consistent with the suggestion that generation of ventilatory components in the heart rate variability (HRV)signal are dependent on spatially distinct loci for cardiac VPN. Therefore,this study has demonstrated the presence of RSA in the HRV signal and a dual location for VPN in the rattlesnake. We suggest there to be a causal relationship between these two observations.