Does physiological tolerance to acute hypoxia and salinity change explain ecological niche in two intertidal crab species?

Intertidal biota is subjected to significant fluctuations in environmental parameters such as salinity and dissolved oxygen (DO). In the current study, the effects of salinity and DO on metabolic rate, critical oxygen partial pressure (P crit), heart rate and osmoregulation in two intertidal crab species commonly found on New Zealand coastlines, Hemigrapsus crenulatus and Hemigrapsus sexdentatus, were measured. Based on its habitation of burrows in the lower intertidal zone, H. crenulatus was predicted to be more resilient to these environmental stressors than H. sexdentatus, which is distributed in the mid to high tidal zone. However, relative to the full-strength seawater control, there were no consistent salinity-dependent changes in respiratory or cardiovascular endpoints in either species following acute 6-h exposures mimicking a tidal cycle. Analysis of haemolymph osmolality and ions determined that both crab species were strong osmotic and ionic regulators over the 6-h exposure period. However, the threshold salinities at which significant changes in osmotic and ionic regulation occurred did differ and generally indicated that H. crenulatus was the better regulator. Respiratory and cardiovascular responses to DO were prominent, with a strong bradycardia observed in both species. Changes in osmolality and sodium ion regulation were also seen as DO declined. The effect on sodium ion levels had its onset at a higher oxygen partial pressure in H. sexdentatus than in H. crenulatus, indicative of a relatively poorer hypoxia tolerance in the former species. The relative resilience of respiratory, cardiovascular and osmoregulatory processes to salinity and DO variations likely contribute to distinct habitat distributions of the two crab species on New Zealand shorelines, although behaviour and inter-specific interactions may also play important roles. Environmental change, in the form of coastal erosion and anthropogenic contamination of estuaries, has the potential to disturb the delicate niche separation that exists between these species.

Behavioural, physiological and biochemical responses to aquatic hypoxia in the freshwater crayfish, Paranephrops zealandicus.

Hypoxia resulting from aquatic eutrophication threatens the population health of the New Zealand freshwater crayfish (koura), Paranephrops zealandicus. An integrated study, combining behavioural, physiological and biochemical approaches, was therefore conducted to characterise the tolerance of this species to hypoxia. When provided with a choice between water flows of high or low dissolved oxygen in short-term laboratory assays, crayfish did not preferentially inhabit waters of higher PO2. However, when an aerial refuge was provided and dissolved oxygen was progressively decreased, crayfish emersed at a PO2 of 0.56±0.03kPa, suggesting a relatively high tolerance to hypoxia. Closed-box respirometry delineated a Pcrit, the point at which crayfish transition from oxyregulating to oxyconforming, of 6.0kPa. Simultaneous measurement of heart rate showed no changes across the PO2 range. In response to 6-h exposures to fixed dissolved oxygen levels (normoxia, 19.3kPa; moderate hypoxia, 3.5kPa; and severe hypoxia, 1.7kPa), P. zealandicus showed a haemolymph PO2 that declined with the magnitude of hypoxia, and while plasma pH declined in severe hypoxia, there were no changes in plasma PCO2. Plasma glucose concentrations fell, and plasma lactate increased in both hypoxic groups. There were no changes in tissue glucose or lactate concentrations. These data indicate that P. zealandicus is relatively tolerant of hypoxia, and possesses biochemical and physiological mechanisms that facilitate survival during short-term exposures to acute hypoxia. If hypoxia is severe and/or prolonged, then this species is capable of escaping to aerial refugia.

Physiological and biochemical strategies for withstanding emersion in two galaxiid fishes.

The galaxiid fishes of the Southern hemisphere display variable tolerance to aerial exposure. Brown mudfish (Neochanna apoda), for example, pseudoaestivate, inhabiting moist soil for months at a time, whereas inanga (Galaxias maculatus) emerse under unfavourable water conditions, but only for periods of a few hours. This study sought to identify the physiological and biochemical strategies that determine emersion tolerance in these species. Nitrogenous waste excretion was measured before and after an experimental emersion period (14 days for mudfish, 6 h for inanga). Both species showed significantly elevated ammonia "washout" upon return to water, but no increase in plasma or muscle ammonia. Post-emersion urea levels were elevated in plasma and muscle in both fish, however the extent of the accumulation did not indicate significant de novo urea production. This was supported by the lack of carbamoyl phosphate synthetase activity in tissues. Consequently, mudfish metabolism was examined to determine whether changes in parameters such as oxygen consumption, carbon dioxide excretion, and/or altered metabolic costs (represented by the key ionoregulatory enzyme Na(+), K(+)-ATPase; NKA) could explain emersion tolerance. Oxygen consumption rates, already very low in immersed mudfish, were largely maintained over the course of emersion. Carbon dioxide excretion decreased during emersion, and a small, but significant, decrease in NKA was noted. These data suggest that the extended emersion capacity of mudfish may result from a generally low metabolic rate that is maintained throughout aerial exposure via cutaneous gas exchange, and which limits the production of potentially toxic nitrogenous waste.

Is the habitation of acidic-water sanctuaries by galaxiid fish facilitated by natural organic matter modification of sodium metabolism?

Acidic waters of New Zealand's West Coast are hypothesized to be a refuge for native galaxiid fish, allowing them to escape predation from acid-sensitive invasive salmonid species. To determine the mechanisms by which galaxiids tolerate low pH, we investigated sodium metabolism in inanga Galaxias maculatus in response to water pH, short-term acclimation to acidic waters, the presence and source of natural organic matter (NOM), and fish life history. Contrary to expectation, inanga were physiologically sensitive to acid exposure, displaying inhibited sodium influx and exacerbated sodium efflux. Short-term (144 h) acclimation to acid did not modify this effect, and NOM did not exert a protective effect on sodium metabolism at low pH. Inanga sourced from naturally acidic West Coast waters did, however, display a sodium influx capacity (J(max)) that was significantly elevated when compared with that of fish collected from neutral waters. All inanga, independent of source, exhibited exceptionally high sodium uptake affinities (18-40 µM) relative to previously studied freshwater teleosts. Although inanga displayed relatively poor physiological tolerance to acidic waters, their high sodium influx affinity coupled with their occupation of near-coastal waters with elevated sodium levels may permit habitation of low-pH freshwaters.

Albuminuria increases cystatin C excretion: implications for urinary biomarkers.

BACKGROUND: Low-molecular weight (LMW) proteins, including albumin and novel urinary biomarkers of acute kidney injury (AKI) such as cystatin C and neutrophil gelatinase-associated lipocalin (NGAL), are normally absorbed from the glomerular filtrate by receptor-mediated transport. We evaluated the effect of albuminuria on urinary excretion of novel biomarkers. METHODS: Sprague-Dawley rats given four injections over 2 days of 5 mg/g body wt/day bovine serum albumin (BSA) in saline were compared with controls given saline alone. Urinary cystatin C, albumin and protein excretion rates were compared prior to treatment (Day -1), after treatment (Day 2) and 4 days later (Day 6). A preliminary assessment of the clinical effect of proteinuria on the filtered urinary biomarkers cystatin C and NGAL was made by comparison with the effect on urinary interleukin-18 (IL-18) that is not absorbed from the glomerular filtrate, in a cohort of intensive care unit patients. RESULTS: BSA induced transient increases in albuminuria, proteinuria and cystatinuria (P < 0.01, P < 0.001 and P < 0.001, respectively). Beyond a threshold 6-fold increase in albuminuria, cystatin C absorption was reduced by competitive inhibition. The excretion rates of all analytes returned to preinjection levels by Day 6. Clinical proteinuria was associated with increasing cystatin C and NGAL concentrations (n = 90, P < 0.0001) but not IL-18 (P = 0.12). CONCLUSIONS: Proteinuria may increase the threshold for detection of AKI by increasing the excretion of LMW protein biomarkers.

Modelling acute renal failure using blood and breath biomarkers in rats.

This paper compares three methods for estimating renal function, as tested in rats. Acute renal failure (ARF) was induced via a 60-min bilateral renal artery clamp in 8 Sprague-Dawley rats and renal function was monitored for 1 week post-surgery. A two-compartment model was developed for estimating glomerular filtration via a bolus injection of a radio-labelled inulin tracer, and was compared with an estimated creatinine clearance method, modified using the Cockcroft-Gault equation for rats. These two methods were compared with selected ion flow tube-mass spectrometry (SIFT-MS) monitoring of breath analytes. Determination of renal function via SIFT-MS is desirable since results are available non-invasively and in real time. Relative decreases in renal function show very good correlation between all 3 methods (R²=0.84, 0.91 and 0.72 for breath-inulin, inulin-creatinine, and breath-creatinine correlations, respectively), and indicate good promise for fast, non-invasive determination of renal function via breath testing.

Differential contribution of diabetes and the Ren2 gene to glomerular pathology in diabetic (mREN-2)27 rats.

The effect of diabetes mellitus vs the effect of the Ren2 gene on the glomerular pathology of (mREN-2)27 heterozygous male rats is controversial. As discrete diabetes-induced glomerular lesions may have been overlooked, we performed a detailed morphometric analysis of glomeruli in diabetic and non-diabetic heterozygous male (mREN-2)27 rats and their normotensive (non-diabetic and diabetic Sprague-Dawley) controls. Glomeruli were scored by light microscopy for nine discrete histological parameters, some of which were graded for extent and/or severity. Mesangiolysis, segmental hypocellularity, and severe tuft-to-capsule adhesions were specific to diabetes; severe mesangial matrix expansion, glomerulosclerosis, thickening of Bowman's capsule, and dilatation of the urinary space were specific to the Ren2 gene. Hyalinosis and hypercellularity were associated with both diabetes and the Ren2 gene: the effect was additive for hyalinosis and synergistic for hypercellularity. The histological parameters were then combined with two physiological indices (systolic blood pressure and proteinuria) and principle components analysis (PCA) was used to detect correlations between the variables. Four discrete patterns of pathology were identified; three were statistically associated with diabetes and/or the Ren2 gene. These findings suggest that both diabetes and the Ren2 gene make significant, albeit different, contributions to the glomerular pathology of diabetic heterozygous male (mREN-2)27 rats. Despite defining the contribution of diabetes, our work does not support the (mREN-2)27 rat as a model of diabetic nephropathy (DN). Rather, it suggests that these animals remain useful for investigating a particular and limited constellation of DN features.

Control of branchial artery tone in fish: effects of environmental temperature and phylogeny.

Branchial artery vasoreactivity and nerve density were measured in teleosts of different phylogenetic relatedness from cold (-1.86 degrees to +1 degrees C) and temperate ( approximately 13 degrees C) environments. Polar Notothenia coriiceps and temperate Paranotothenia angustata are closely related Southern Hemisphere nototheniids, and polar Boreogadus saida and temperate Myoxocephalus scorpius are Northern Hemisphere species that are phylogenetically distant from each other and the nototheniids. Rainbow trout Oncorhynchus mykiss was used as a temperate comparison of different activity patterns. Cumulative dose-response curves for efferent branchial arteries (EBAs) revealed a lack of alpha-adrenergic (noradrenaline [NOR]) vasoactivity in M. scorpius and a lack of cholinergic (carbachol [CARB]) vasoactivity in B. saida, while responses in N. coriiceps and P. angustata were irregular. Oncorhynchus mykiss vessels were responsive to both constrictors, suggesting either receptor absence or spatial heterogeneity among species. The sensitivity (pEC(50)) of NOR or 5-hydroxytryptamine (5-HT) vasoconstriction showed little interspecific variation, while maximal tension varied among species for NOR but not 5-HT. Acute cooling did not affect agonist-independent vascular contraction induced by KCl or in response to NOR or 5-HT. The CARB sensitivity of EBAs increased with cooling in P. angustata alone. Each species possessed a unique pattern of innervation on both branchial arteries, distal and proximal to their respective aortas, that partially differentiated between nototheniids and nonnototheniids. We conclude there is little evidence for cold adaptation of branchial artery contractility, and mechanisms of vascular control likely reflect phylogeny rather than thermal history.

Effects of gill abrasion and experimental infection with Tenacibaculum maritimum on the respiratory physiology of Atlantic salmon Salmo salar affected by amoebic gill disease.

The effects of gill abrasion and experimental infection with Tenacibaculum maritimum were assessed in Atlantic salmon Salmo salar with underlying amoebic gill disease. The respiratory and acid-base parameters arterial oxygen tension (P(a)O2), arterial whole blood oxygen content (C(a)O2), arterial pH (pHa), haematocrit and haemoglobin concentrations were measured at intervals over a 48 h recovery period following surgical cannulation of the dorsal aorta. Mortality rates over the recovery period were variable, with gill abrasion and inoculation with T. maritimum causing the highest initial mortality rate and unabraded, uninoculated controls showing the lowest overall mortality rate. Fish with abraded gills tended to show reduced P(a)O2 and lower C(a)O2 compared with unabraded fish. Infection with T. maritimum had no effect on P(a)O2 or C(a)O2. All fish showed an initial alkalosis at 24 h post-surgery/inoculation which was more pronounced in fish inoculated with T. maritimum. There were no significant effects of gill abrasion or infection upon the ratio of oxygen specifically bound to haemoglobin or mean cellular haemoglobin concentration. Histologically, 48 h following surgery, abraded gills showed multifocal hyperplastic lesions with pronounced branchial congestion and telangiectasis, and those inoculated with T. maritimum exhibited focal areas of branchial necrosis and erosion associated with filamentous bacterial mats. All fish examined showed signs of amoebic gill disease with multifocal hyperplastic and spongious lesions with parasome-containing amoeba associated with the gill epithelium. The results suggest that respiratory compromise occurred as a consequence of gill abrasion rather than infection with T. maritimum.

Cardiovascular responses of Chinook salmon (Oncorhynchus tshawytscha) during rapid anaesthetic induction and recovery.

The effects of three anaesthetics on induction and recovery were compared in Chinook salmon (Oncorhynchus tshawytscha). Heart rate (HR), cardiac output (Q), dorsal aortic pressure (DAP) and stroke volume (SV) were measured in minimally disturbed salmon during 5 min anaesthetic inductions with approximately equi-potent concentrations of MS222 (100 ppm), metomidate (6-10 ppm) and AQUI-S (60 ppm). MS222 induction caused a steady decline in DAP only, while metomidate induction did not affect any cardiovascular variable. AQUI-S caused a biphasic response, and within 2 min had depressed HR, Q, DAP and SV by between 20 and 50%. In the final 3 min HR returned to pre-anaesthesia levels, and Q and SV climbed to greater than pre-anaesthesia levels. Blood samples taken pre- and post-anaesthesia showed all inductions caused hypoxaemia (oxygen partial pressure of dorsal aortic blood (PaO2): MS222 47 mmHg, metomidate 35 mmHg, AQUI-S 21 mmHg). Haematocrit and plasma adrenaline and noradrenaline levels increased slightly in AQUI-S treated fish only. Recovery was monitored for 6 h post-anaesthesia, and was similar for each anaesthetic. All cardiovascular variables had returned to control levels within 5 min with the exception of DAP, which was initially slightly elevated (up to 20%) but returned to control values within 30 min. Anaesthesia is usually preceded by handling. Netting prior to anaesthesia caused significant increases in HR, Q and SV, which masked any anaesthetic dependent effects. Recovery from anaesthesia combined with surgery was also generally anaesthetic independent and recovery was prolonged, compared to anaesthesia alone. These data suggest limiting fish handling/manipulation is more important in minimising cardiovascular disturbance than the choice of anaesthetic.