Showing papers in "Comparative Biochemistry and Physiology A-molecular & Integrative Physiology in 2010"

Starvation physiology: reviewing the different strategies animals use to survive a common challenge.

Abstract: All animals face the possibility of limitations in food resources that could ultimately lead to starvation-induced mortality. The primary goal of this review is to characterize the various physiological strategies that allow different animals to survive starvation. The ancillary goals of this work are to identify areas in which investigations of starvation can be improved and to discuss recent advances and emerging directions in starvation research. The ubiquity of food limitation among animals, inconsistent terminology associated with starvation and fasting, and rationale for scientific investigations into starvation are discussed. Similarities and differences with regard to carbohydrate, lipid, and protein metabolism during starvation are also examined in a comparative context. Examples from the literature are used to underscore areas in which reporting and statistical practices, particularly those involved with starvation-induced changes in body composition and starvation-induced hypometabolism can be improved. The review concludes by highlighting several recent advances and promising research directions in starvation physiology. Because the hundreds of studies reviewed here vary so widely in their experimental designs and treatments, formal comparisons of starvation responses among studies and taxa are generally precluded; nevertheless, it is my aim to provide a starting point from which we may develop novel approaches, tools, and hypotheses to facilitate meaningful investigations into the physiology of starvation in animals.

Metabolic characteristics and oxidative damage to skeletal muscle in broiler chickens exposed to chronic heat stress.

Abstract: Emerging evidence has shown that acute heat exposure affects metabolic characteristics and causes oxidative damage to skeletal muscle in birds. Little is known, however, about such phenomena under chronic heat stress conditions. To address this, we designed the present study to determine the influence of cyclic (32 to 24 to 32 degrees C: 32 degrees C for 8 h/d, 32-24-32HS ), and constant (32 and 34 degrees C, 32HS and 34HS, respectively) heat exposure on the metabolic and peroxide status in skeletal muscle of 4-wk-old male broiler chickens. Heat stress, particularly in the 32HS and 34HS groups, depressed feed intake and growth, while cyclic high temperature gave rise to a less severe stress response in performance terms. Malondialdehyde (MDA) levels in skeletal muscle were enhanced (P<0.05) by constant heat treatment; the degree of enhancement was not as large as the changes observed in our previous 'acute' heat stress model. The 3HADH (3-hydroxyacyl CoA dehydrogenase related to fatty acid oxidation) and CS (citrate synthase) enzyme activities were lowered (P<0.05) by both the cyclic and constant 34HS treatments, and constant 34HS group, respectively. These results suggest that chronic heat exposure decreases metabolic oxidation capacity in skeletal muscle of broiler chickens. On exposure to chronic heat stress, GPx activity remained relatively constant, though a temperature-dependent elevation in Cu/Zn-SOD activity was observed, implying that anti-oxidation ability was disturbed by the chronic stress condition. From these results it can be concluded that chronic heat stress did not induce oxidative damage to a major extent. This may probably be due to a decrease in metabolic oxidation capacity or due to a self-propagating scavenging system, though the system was not fully activated.

δ15N and δ13C diet-tissue discrimination factors for large sharks under semi-controlled conditions

Abstract: Stable isotopes ( δ 15 N and δ 13 C) are being widely applied in ecological research but there has been a call for ecologists to determine species- and tissue-specific diet discrimination factors (∆ 13 C and ∆ 15 N) for their study animals. For large sharks stable isotopes may provide an important tool to elucidate aspects of their ecological roles in marine systems, but laboratory based controlled feeding experiments are impractical. By utilizing commercial aquaria, we estimated ∆ 15 N and ∆ 13 C of muscle, liver, vertebral cartilage and a number of organs of three large sand tiger ( Carcharias taurus ) and one large lemon shark ( Negaprion brevirostris ) under a controlled feeding regime. For all sharks mean ± SD for ∆ 15 N and ∆ 13 C in lipid extracted muscle using lipid extracted prey data were 2.29‰ ± 0.22 and 0.90‰ ± 0.33, respectively. The use of non-lipid extracted muscle and prey resulted in very similar ∆ 15 N and ∆ 13 C values but mixing of lipid and non-lipid extracted data produced variable estimates. Values of ∆ 15 N and ∆ 13 C in lipid extracted liver and prey were 1.50‰ ± 0.54 and 0.22‰ ± 1.18, respectively. Non-lipid extracted diet discrimination factors in liver were highly influenced by lipid content and studies that examine stable isotopes in shark liver, and likely any high lipid tissue, should strive to remove lipid effects through standardising C:N ratios, prior to isotope analysis. Mean vertebral cartilage ∆ 15 N and ∆ 13 C values were 1.45‰ ± 0.61 and 3.75‰ ± 0.44, respectively. Organ ∆ 15 N and ∆ 13 C values were more variable among individual sharks but heart tissue was consistently enriched by ~ 1–2.5‰. Minimal variability in muscle and liver δ 15 N and δ 13 C sampled at different intervals along the length of individual sharks and between liver lobes suggests that stable isotope values are consistent within tissues of individual animals. To our knowledge, these are the first reported diet–tissue discrimination factors for large sharks under semi-controlled conditions, and are lower than those reported for teleost fish.

Effect of heat stress on the porcine small intestine: a morphological and gene expression study.

Abstract: With the presence of global warming, the occurrence of extreme heat is becoming more common, especially during the summer, increasing pig susceptibility to severe heat stress. The aim of the current study was to investigate changes in morphology and gene expression in the pig small intestine in response to heat stress. Forty eight Chinese experimental mini pigs (Sus scrofa) were subjected to 40 degrees C for 5h each day for 10 successive days. Pigs were euthanized at 1, 3, 6, and 10 days after heat treatment and sections of the small intestine epithelial tissue were excised for morphological examination and microarray analyses. After heat treatment, the pig rectal temperature, the body surface temperature and serum cortisol levels were all significantly increased. The duodenum and jejunum displayed significant damage, most severe after 3 days of treatment. Microarray analysis found 93 genes to be up-regulated and 110 genes to be down-regulated in response to heat stress. Subsequent bioinformatic analysis (including gene ontology and KEGG pathway analysis) revealed the genes altered in response to heat stress related to unfolded protein, regulation of translation initiation, regulation of cell proliferation, cell migration and antioxidant regulation. Heat stress caused significant damage to the pig small intestine and altered gene expression in the pig jejunum. The results of the bioinformatic analysis from the present study will be beneficial to further investigate the underlying mechanisms involved in heat stress-induced damage in the pig small intestine.

Effects of elevated temperature on coral reef fishes: Loss of hypoxia tolerance and inability to acclimate

Abstract: Water temperature is expected to rise on coral reefs due to global warming. Here, we have examined if increased temperature reduces the hypoxia tolerance of coral reef fish (measured as critical [O(2)]), and if temperature acclimation in adults can change the resting rate of O(2) consumption and critical [O(2)]. Two common species from Lizard Island (Great Barrier Reef, Australia) were tested, Doederlein's cardinalfish (Ostorhinchus doederleini) and lemon damselfish (Pomacentrus moluccensis). In both species, a 3 degrees C rise in water temperature caused increased oxygen consumption and reduced hypoxia tolerance, changes that were not reduced by acclimation to the higher temperature for 7 to 22 days. Critical [O(2)] increased by 71% in the cardinalfish and by 23% in the damselfish at 32 degrees C compared to 29 degrees C. The higher oxygen needs are likely to reduce the aerobic scope, which could negatively affect the capacity for feeding, growth and reproduction. The reduced hypoxia tolerance may force the fishes out of their nocturnal shelters in the coral matrix, exposing them to predation. The consequences for population and species survival could be severe unless developmental phenotypic plasticity within generations or genetic adaptation between generations could produce individuals that are more tolerant to a warmer future.

Response of Mytilus galloprovincialis (L.) to increasing seawater temperature and to marteliosis: metabolic and physiological parameters.

Abstract: In the context of climate change the present work aimed to illustrate whether the energetic and metabolic pattern of mussels Mytilus galloprovincialis will be affected by increase in the temperature of seawater. Moreover we examined whether an outbreak of Marteilia sp. infestation as a result of increase in sea water temperature will impair the energetic balance of mussels. M. galloprovincialis was acclimated at 18 degrees C, 24 degrees C, 26 degrees C and 28 degrees C for 30 days and the energetic pattern of its tissues was estimated by determining the factor Scope for Growth (SFG), while the metabolic pattern of mussels was estimated by determining the activities of pyruvate kinase (PK) and phosphoenolpyruvate carboxykinase (PEPCK). The decrease in PK activity and the decrease in the ratio PK/PEPCK indicated an activation of anaerobic component of metabolism during acclimation of mussels at temperature 24 degrees C. At temperatures higher than 24 degrees C the values of SFG turned negative probably associated with a significant reduction in clearance rate. Compared to the non infected mussels, the SFG values of infected mussels were significantly lower (P<0.05). These differences were attributed to the higher filtration rate and the lower absorption efficiency detected in the infected mussels. Also the degree of SFG reduction is dependent on the intensity levels of infection by Marteilia sp.

Perspectives on carbonic anhydrase

Abstract: In the years since Larimer and Schmidt-Nielsen published their examination of red blood cell (RBC) carbonic anhydrase (CA) activities as a function of body mass in mammals, our knowledge of CA has expanded dramatically. We are now aware of the diversity of CA isoforms and their implication in a wide array of physiological processes. The catalytic mechanism of CA has been described, and numerous compounds that function as activators or inhibitors of CA activity have been identified. CA is investigated as a diagnostic tumor marker, and CA inhibitors are used or emerging as clinical treatments for diseases as diverse as glaucoma, cancer and obesity. Yet despite the intensity of research effort over the last 50years and the wealth of information that has accumulated, the questions asked by Larimer and Schmidt-Nielsen remain relevant today - we still have much to learn about the patterns and physiological significance of interspecific differences in CA expression and activity.

Immune responses and expression of immune-related genes in swimming crab Portunus trituberculatus exposed to elevated ambient ammonia-N stress.

Abstract: The effects of ammonia-N (0, 1, 5 and 20mgL(-1)) on immune responses and immune-related gene expression were determined in swimming crab Portunus trituberculatus. The results showed that the THC and phagocytic activity of P. trituberculatus exposed to 1, 5 and 20mgL(-1) ammonia-N decreased significantly during the experimental time. The antibacterial and bacteriolytic activities decreased significantly to the minimum at 6 or 12h respectively, then recovered to the control level except bacteriolytic activity exposed to 5 and 20mgL(-1) ammonia-N. alpha(2)-macroglobulin (alpha(2)-M) activity of all ammonia-N exposure groups decreased significantly, then recovered to the control level rapidly and tended to be stable after 12h. With crabs exposed to 20mgL(-1) ammonia-N, the gene expression levels of crustin and lysozyme decreased significantly, then recovered to the control level after 12h. ALF expression also decreased significantly when exposed to 5 and 20mgL(-1) ammonia-N, then remained stable and significantly lower than the control group after 6h. In contrast, alpha(2)-M gene expression was induced by ammonia-N exposure significantly. The results suggest that high concentration ammonia-N exposure could reduce the crab immunity severely, and induce the short-term response in terms of immune gene regulation.

Rapid changes in plasma cortisol, osmolality, and respiration in response to salinity stress in tilapia (Oreochromis mossambicus).

Abstract: We elucidated a time course for cortisol release in tilapia as it corresponds to changes in plasma osmolytes and respiration. Following exposure of freshwater (FW) tilapia to 25 per thousand seawater (SW), we measured plasma osmolality, [Na(+)], [K(+)], [Cl(-)], hematocrit, cortisol concentration, oxygen-consumption rate (MO2), and ventilation frequency over 5days and compared them to FW control fish. Cortisol increased rapidly by 3h and remained elevated for 3days. Plasma osmolality, [Na(+)], and [Cl(-)] were elevated at 6-8h, peaked 24h following SW exposure, and then decreased to near-FW levels by 3days. MO2 increased at 24h post-SW exposure relative to FW, while ventilation frequency increased by 3h. Overall, we interpret changes in cortisol as resulting from a change in salinity, in contrast to changes in plasma solute concentrations that could be due to adjustments resulting from the fish's cortisol response as it faces osmoregulatory distress. Increases in oxygen-consumption rate at 24h and ventilation frequency at 3h are likely as a result of the cellular stress response occurring during salinity stress. No significant changes in blood hematocrit were observed, which suggests that tilapia are capable of rapidly counteracting dehydration during acute hyperosmotic stress.

Long-term warm or cold acclimation elicits a specific transcriptional response and affects energy metabolism in zebrafish

Abstract: Organisms are often forced to acclimate to changing environmental temperature. Temperature compensation mechanisms have been reported, which enable organisms to minimize some of the temperature related effects. To investigate this process, zebrafish (Danio rerio) were acclimated to a control (26 °C), an increased (34 °C) or a decreased (18 °C) temperature for 4, 14 and 28 days. In general, warm acclimation depleted energy stores and decreased the condition factor, while cold acclimation increased both. The energy parameters as well as the transcriptional responses (investigated using printed 15k microarrays and real time PCR) indicated that warm acclimation was particularly stressful. However, after 28 days of warm acclimation, energy stores had recovered from the initial depletion. This could have been facilitated by the observed downregulation of transcripts involved in catabolic processes. Transcriptional regulation seemed to be an important means of coordinating the temperature compensation process. We could distinguish an early response which was independent of the direction of the temperature change and a direction specific long-term response. The early response was characterized by the upregulation of defence mechanisms, tissue regeneration and hemopoiesis. In the long-term response there was a strong emphasis on compensating for the altered metabolic rate as well as cell structure and replacement.

The effect of plant protein-based diet supplemented with dipeptide or free amino acids on digestive tract morphology and PepT1 and PepT2 expressions in common carp (Cyprinus carpio L.).

Abstract: Common carp (Cyprinus carpio) of average body mass 0.07+/-0.02 g were fed three formulated diets: wheat gluten protein-based diet supplemented with Lys-Gly dipeptide (PP), wheat gluten protein-based diet supplemented with free lysine and glycine (AA), and a wheat gluten protein-based control diet without lysine supplementation (CON), frozen zooplankton (Z) (restricted diet), and a commercial starter food Aglo Norse (AN). After 4 weeks of experimental feeding, fish fed AN diet showed the highest body mass and length. Significantly lower mass occurred in groups fed PP, AA, CON, and Z. Fish fed CON diet showed the lowest intestinal folds and the highest number of mucous cells. Fish fed PP diet showed a significantly higher number of gastrin/cholecystokinin (CCK) positive cells. The diameter of lipid vacuoles in hepatocyte cytoplasm of fish fed formulated diets (PP, AA and CON) was significantly higher than in fish fed zooplankton (Z) and the commercial diet (AN). Hepatocytes of fish fed AA and CON showed a higher nucleus proliferation rate than in the other experimental groups. The quantitative analysis of the number of proliferating cell nuclear antigen (PCNA) and caspase-3(rabbit polyclonal antibody CPP-32)-positive cells showed that the highest proliferation rate was accompanied by the high apoptosis in the intestine of fish fed AA and CON. After 4 weeks of experimental feeding the highest relative expression of PepT1 gene was observed in fish fed PP diet, while the lowest expression occurred in fish fed CON. Feeding carp plant protein-based diet supplemented with Lys-Gly dipeptide (PP) had a beneficial influence on fish growth and metabolism in the digestive tract as compared to fish fed control diet without lysine supplementation (CON).

The unusual energy metabolism of elasmobranch fishes.

Abstract: article i nfo Article history: The unusual energy metabolism of elasmobranchs is characterized by limited or absent fatty acid oxidation in cardiac and skeletal muscle and a great reliance on ketone bodies and amino acids as oxidative fuels in these tissues. Other extrahepatic tissues in elasmobranchs rely on ketone bodies and amino acids for aerobic energy production but, unlike muscle, also appear to possess a significant capacity to oxidize fatty acids. This organization of energy metabolism is reflected by relatively low plasma levels of non-esterified fatty acids (NEFA) and by plasma levels of the ketone body s-hydroxybutyrate that are as high as those seen in fasted mammals. The preference for ketone body oxidation rather than fatty acid oxidation in muscle of elasmobranchs under routine conditions is opposite to the situation in teleosts and mammals. Carbohydrates appear to be utilized as a fuel source in elasmobranchs, similar to other vertebrates. Amino acid- and lipid- fueled ketogenesis in the liver, the lipid storage site in elasmobranchs, sustains the demand for ketone bodies as oxidative fuels. The liver also appears to export NEFA and serves a buoyancy role. The regulation of energy metabolism in elasmobranchs and the effects of environmental factors remain poorly understood. The metabolic organization of elasmobranchs was likely present in the common ancestor of the Chondrichthyes ca. 400million years ago and, speculatively, it may reflect the ancestral metabolism of jawed vertebrates. We assess hypotheses for the evolution of the unusual energy metabolism of elasmobranchs and propose that the need to synthesize urea has influenced the utilization of ketone bodies and amino acids as oxidative fuels.

Dietary lipid composition and avian migratory flight performance: Development of a theoretical framework for avian fat storage

Abstract: Birds rely substantially on fat to fuel migratory flights. The importance of the composition of those fat stores to flight performance is a field of recent interest. Here I review the evidence that dietary lipid fat composition affects exercise in birds. Seasonal changes in adipose composition and diet choice experiments have yet to provide strong evidence that fatty acid composition can affect flight performance. Direct manipulations of dietary fat, however, have been demonstrated to affect exercise performance in both avian and non-avian species. I also describe the major hypotheses for the mechanisms by which dietary fat affects exercise, focusing on the role of fatty acids as oxidative substrates and as structural components of membranes. Evidence is accumulating that fatty acids that are shorter or have more double bonds increase peak performance due to their higher transport rates en route to oxidation. Endurance and efficiency of flight may or may not be affected in similar ways. Other mechanisms requiring further investigation include membrane composition, peroxisome proliferator-activated receptors, and eicosanoid-mediated inflammation. Finally, I develop a theoretical framework for studying the composition of fat stores in migrants, focusing on the tradeoffs between fatty acid transport rates, energy storage, and assimilation during stopover refueling.

The effect of peptide absorption on PepT1 gene expression and digestive system hormones in rainbow trout (Oncorhynchus mykiss).

Abstract: The present study evaluates the effect of protein source (dipeptides, free amino acids, and intact protein) on development and growth of Salmonid fish alevin. Specifically, we follow the expression of oligopeptide transporter protein PepT1 in the intestine of rainbow trout ( Oncorhynchus mykiss ). Fish were fed exogenously one of four diets: three formulated (lysyl–glycine dipeptide supplemented diet — PP, free lysine and glycine supplemented diet — AA, control diet with no lysine — CON) or commercial starter (Aller Futura — AF). Fish increased mean body weight 8 fold with PP- and AA-supplemented diets resulting in significantly higher weight gain than fish fed CON. Statistical analysis revealed a significant increase in relative PepT1 expression of fish fed experimental diets. Immunohistochemical staining with PepT1 antibody showed the presence of the transporter protein in the brush border membrane of the proximal intestinal enterocytes of fish from all experimental groups. Leptin immunoreactivity occurred not only in the gastric glands but also in proximal intestine and pyloric caeca of fish fed PP, AA and AF diets. Leptin immunoreactivity was also observed in hepatocyte cytoplasm and pancreatic acinar cells. Gastrin/CCK immunoreactive cells were present in the proximal intestine and pyloric caeca.

Perspective--Exercise in fish: 50+years and going strong.

Abstract: Swimming in fish has garnered the attention of researchers for more than 50years. Research has focused on the mechanisms that fish utilize during exercise, fuel use patterns, recovery dynamics and the effects of abiotic and biotic factors on fish exercise physiology. Research direction has been influenced both by technology (e.g., availability of telemetry devices and access to swim flumes) and environmental changes (e.g., global warming and pollution). In addition, researchers have begun to examine the interplay between swimming physiology and behaviour. As part of the 50th Anniversary of Comparative Physiology and Biochemistry, this perspective examines the history of fish exercise research, and some of the individuals who have made significant contributions.

Physiological response of the blue mussel Mytilus edulis to differences in food and temperature in the Gulf of Maine.

Abstract: Studies performed in the Gulf of Maine (GOM) during the spring of 2006 examined populations of the blue mussel, Mytilus edulis at several intertidal locations. Several parameters were measured including maximum length, diet (stable isotope composition), and the physiological performance of individual mussels using condition indices and RNA/DNA ratios. These same mussels were also assessed for their response to differences in seawater temperature by quantifying the expression of heat shock proteins (HSP70) and the activities of the antioxidant enzyme, superoxide dismutase (SOD). These data were then interpreted in the context of sea surface and air temperatures as well as chlorophyll a concentration and the genetic structure of mussel populations. Populations of M. edulis throughout the GOM were found to be genetically homogenous and consumed a mixed diet of phytoplankton and detritus. Mussels exposed to higher seawater temperatures also showed a significant increase in the expression of HSP70 and activities of SOD. The site-specific interplay between the amount of energy gained from the available food resources and the costs associated with protection against the effects of elevated seawater temperatures shows that these mussels exhibit phenotypic plasticity at different sites which could play an important role in the population dynamics of this key member of the rocky intertidal.

Heat shock protein gene expression and function in amphibian model systems.

Abstract: Heat shock proteins (HSPs) are molecular chaperones that are involved in protein folding and translocation. During heat shock, both constitutive and stress-inducible HSPs bind to and inhibit irreversible aggregation of denatured protein and facilitate their refolding once normal cellular conditions are re-established. Recent interest in HSPs has been propelled by their association with various human diseases. Amphibian model systems, as shown in this review, have had a significant impact on our understanding of hsp gene expression and function. Some amphibian hsp genes are expressed constitutively during oogenesis and embryogenesis, while others are developmentally regulated and enriched in selected tissues in a stress-inducible fashion. For example, while hsp70 genes are heat-inducible after the midblastula stage, hsp30 genes are not inducible until late neurula/early tailbud. This particular phenomenon is likely controlled by chromatin structure. Also, hsp genes are expressed during regeneration, primarily in response to wounding-associated trauma. The availability of amphibian cultured cells has enabled the analysis of hsp gene expression induced by different stresses (e.g. cadmium, arsenite, proteasome inhibitors etc.), HSP intracellular localization, and their involvement in stress resistance. Furthermore, hyperthermia treatment of adult amphibians reveals that certain tissues were more sensitive than others in terms of hsp gene expression. Finally, this review details the evidence available for the role of amphibian small HSPs as molecular chaperones.

Salinity-dependent expression of a Na+, K+, 2Cl- cotransporter in gills of the brackish medaka Oryzias dancena: a molecular correlate for hyposmoregulatory endurance.

Abstract: This study used the brackish medaka (Oryzias dancena) to characterize Na+, K+, 2Cl− cotransporter (NKCC) expression from the genetic to cellular level in gills. Using RT-PCR to survey tissue distribution of nkcc1a, 1b, and 2, we report that gills of brackish medaka prominently express Odnkcc1a. The full-length cDNA of Odnkcc1a was cloned from gill tissue. In situ hybridization indicates that Odnkcc1a was localized to mitochondrion-rich (MR) cells. Higher mRNA levels of Odnkcc1a were found in gills from seawater (SW) and brackish water (BW) medaka when compared to freshwater (FW) fish. Furthermore, higher amounts of NKCC1a-like protein were detected by the monoclonal antibody in gills of SW and BW medaka compared to FW medaka. Double immunofluorescence staining revealed that NKCC1a-like protein colocalizes with Na+, K+–ATPase on the basolateral membrane of MR cells in BW and SW fish. In addition, transfer of brackish medaka from SW to FW revealed that expression of NKCC1a-like protein in gills was retained until 7 days, which is a likely mechanism for maintaining hyposmoregulatory endurance. The study illustrates salinity-dependent expression of NKCC1a in branchial MR cells from brackish medaka and suggests a critical role for NKCC1a in hyposmoregulatory endurance of this fish.

Time course of ROS production in skeletal muscle mitochondria from chronic heat-exposed broiler chicken

Abstract: This study was designed to elucidate physiological changes of skeletal muscle mitochondria from broiler chickens (Gallus gallus) during chronic heat exposure. Chickens (19-day-old) were exposed to either constant heat stress (34 degrees C) or kept at control temperature (24 degrees C) for 14days. Mitochondrial ROS production for control group showed little changes during the experimental periods, whereas that for the heat-stressed group was increased after 3, 5 and 9days of heat exposure and returned to original levels at day 14. Mitochondrial membrane potential in state 4 for heat-stressed birds was higher than those of control birds after 3 and 5days, but was not at day 14. Mitochondrial oxygen consumption rate in state 3 was increased after 3 and 5days, and also returned to original levels by day 14. These results suggest that chronic heat stress induces increased ROS production in skeletal muscle mitochondria, probably via elevation of the membrane potential in state 4, resulting from enhanced oxygen consumption in the initial stage of heat exposure. These physiological changes were no longer observed at day 14, possibly because the animals had acclimatized to environmental heat stress.

Claudin-15 and -25b expression in the intestinal tract of Atlantic salmon in response to seawater acclimation, smoltification and hormone treatment.

Abstract: In seawater fishes, osmotic homeostasis requires uptake of ions and water in the intestine and these processes are governed by the combined trans- and paracellular pathways. The current study examined mRNA expression of two tight junction proteins (claudin-15 and -25b) predominantly expressed in the intestine of Atlantic salmon. We examined the response in pyloric caecae, middle and posterior intestine to seawater challenge, during smoltification and after injection with osmoregulatory hormones. Seawater (SW) transfer elevated levels of claudin-15 and -25b while no change was induced throughout the smolt stage. In freshwater, cortisol and growth hormone inhibited claudin-15 expression in the two anterior segments. Claudin-25b was elevated in all intestinal segments by growth hormone, while cortisol had an inhibitory effect. In seawater, prolactin and cortisol inhibited claudin expression. The data suggest that claudin expression is involved in the reorganisation of intestinal epithelium and possibly change paracellular permeability during SW acclimation. The lack of preparatory change during smoltification suggests that this process is not completed during smolt development. The effects of the tested hormones cannot explain the sum of changes induced by salinity, which, like the smoltification data, suggests the importance of additional factors and possibly contact with the imbibed SW per se.

Differential passage of fluids and different-sized particles in fistulated oxen (Bos primigenius f. taurus), muskoxen (Ovibos moschatus), reindeer (Rangifer tarandus) and moose (Alces alces): rumen particle size discrimination is independent from contents stratification.

Abstract: Ruminant species differ in the degree that their rumen contents are stratified but are similar insofar that only very fine particles are passed from the forestomach to the lower digestive tract. We investigated the passage kinetics of fluid and particle markers (2, 10 and 20 mm) in fistulated cattle (Bos primigenius f. taurus), muskoxen (Ovibos moschatus), reindeer (Rangifer tarandus) and moose (Alces alces) on different diets. The distribution of dry matter in the rumen and the viscosity of rumen fluids suggested that the rumen contents were more stratified in muskoxen than moose. Correspondingly, as in previous studies, the species differed in the ratio of mean retention times of small particles to fluids in the reticulorumen, which was highest in cattle (2.03) and muskoxen (1.97-1.98), intermediate in reindeer (1.70) and lowest in moose (0.98-1.29). However, the ratio of large to small particle retention did not differ between the species, indicating similarity in the efficiency of the particle sorting mechanism. Passage kinetics of the two largest particle classes did not differ, indicating that particle retention is not a continuous function of particle size but rather threshold-dependent. Overall, the results suggest that fluid flow through the forestomach differs between ruminant species. A lower relative fluid passage, such as in moose, might limit species to a browse-based dietary niche, whereas a higher relative fluid passage broadens the dietary niche options and facilitates the inclusion of, or specialization on, grass. The function of fluid flow in the ruminant forestomach should be further investigated.

Feast to famine: The effects of food quality and quantity on the gut structure and function of a detritivorous catfish (Teleostei: Loricariidae)

Abstract: The gastrointestinal (GI) tract and associated organs are some of the most metabolically active tissues in an animal. Hence, when facing food shortages or poor food quality, an animal may reduce the size and function of their GI tract to conserve energy. We investigated the effects of prolonged starvation and varying food quality on the structure and function of the GI tract in a detritivorous catfish, Pterygoplichthys disjunctivus, native to the Amazonian basin, which experiences seasonal variation in food availability. After 150 days of starvation or consumption of a wood-diet too low in quality to meet their energetic needs, the fish reduced the surface area of their intestines by 70 and 78%, respectively, and reduced the microvilli surface area by 52 and 27%, respectively, in comparison to wild-caught fish consuming their natural diet and those raised in the laboratory on a high-quality algal diet. Intake and dietary quality did not affect the patterns of digestive enzyme activity along the guts of the fish, and the fish on the low-quality diet had similar mass-specific digestive enzyme activities to wild-caught fish, but lower summed activity when considering the mass of the gut. Overall, P. disjunctivus can endure prolonged starvation and low food quality by down-regulating the size of its GI tract.

The effects of feeding on the swimming performance and metabolic response of juvenile southern catfish, Silurus meridionalis, acclimated at different temperatures.

Abstract: To test whether the effects of feeding on swimming performance vary with acclimation temperature in juvenile southern catfish (Silurus meridionalis), we investigated the specific dynamic action (SDA) and swimming performance of fasting and feeding fish at acclimation temperatures of 15, 21, 27, and 33 degrees C. Feeding had no effect on the critical swimming speeding (U(crit)) of fish acclimated at 15 degrees C (p=0.66), whereas it elicited a 12.04, 18.70, and 20.98% decrease in U(crit) for fish acclimated at 21, 27 and 33 degrees C, respectively (p<0.05). Both the maximal postprandial oxygen consumption rate (VO2peak) and the active metabolic rate (VO2active, maximal aerobic sustainable metabolic rate of fasting fish) increased significantly with temperature (p<0.05). The postprandial maximum oxygen consumption rates during swimming (VO2max) were higher than the VO2active of fasting fish at all temperature groups (p<0.05). The VO2max increased with increasing temperature, but the relative residual metabolic scope (VO2max-VO2peak) during swimming decreased with increasing in temperature. The present study showed that the impairment of postprandial swimming performance increased with increasing temperature due to the unparalleled changes in the catfish's central cardio-respiratory, peripheral digestive and locomotory capacities. The different metabolic strategies of juvenile southern catfish at different temperatures may relate to changes in oxygen demand, imbalances in ion fluxes and dissolved oxygen levels with changes in temperature.

Skeletal muscle cellularity and expression of myogenic regulatory factors and myosin heavy chains in rainbow trout (Oncorhynchus mykiss): Effects of changes in dietary plant protein sources and amino acid profiles

Abstract: The nutritional regulation of skeletal muscle growth is very little documented in fish. The aim of the study presented here was to determine how changes in dietary plant protein sources and amino acid profiles affect the muscle growth processes of fish. Juvenile rainbow trout (Oncorhynchys mykiss) were fed two diets containing fish meal and a mixture of plant protein sources either low (control diet) or rich in soybean meal (diet S). Both diets were supplemented with crystalline indispensable amino acids (IAA) to match the rainbow trout muscle IAA profile. Diet S was also supplemented with glutamic acid, an AA present in high quantities in trout muscle. Rainbow trout fed diets C and S were not significantly different in terms of overall somatic growth or daily nitrogen gain, although their parameters of dietary protein utilisation differed. Distribution of skeletal white muscle fibre diameter and expression of certain selected muscle genes were also affected by dietary changes. In the white muscle, diet S led to a significant decrease (x0.9) in the mean and median diameters of muscle fibres, to a significant decrease (x0.6) in the expression of MyoD and to a significant increase (x1.7) in the expression of fast-MHC, with no significant changes in myogenin expression. There was no change in the expression of the genes analysed in lateral red muscle (MyoD, MyoD2, myogenin and slow-MHC). These results demonstrated that changes occurred in skeletal white muscle cellularity and expression of MyoD and fast-MHC, although overall growth and protein accretion were not modified, when a diet rich in soybean meal and glutamic acid was ingested. Present findings also indicated that the white and red muscles of rainbow trout are differently affected by nutritional changes.

Cave colonization without fasting capacities: an example with the fish Astyanax fasciatus mexicanus.

Abstract: Subterranean animals have commonly evolved hypoactivity, hypometabolism and/or the sequential use of energetic reserves to tolerate long fasting periods imposed by the low food levels found in subterranean environments. However, some tropical caves are characterized by a potential high level of nutriments. By using the tropical fish Astyanax fasciatus that presents both populations subterranean (Astyanaxfasciatus mexicanus) and epigean (Astyanaxfasciatus fasciatus) populations, we described behavioral, metabolic and biochemical responses during a long-term fasting period followed by a refeeding period. The results demonstrated that fed hypogean fishes exhibited different energy stores together with a hypometabolism. But, despite drastic decreases in locomotory activity and oxygen consumption during fasting, hypogean fishes consumed significantly more glycogen, triglycerides and proteins during the starvation period than epigean fishes. This lower fasting capacity showed by hypogean fishes is confirmed by the higher activation of the compensatory metabolic pathways (ketogenesis and gluconeogenesis). After the refeeding period, cave fishes did not recover from the "food deprivation" stress, and resume fed levels in glycogen, triglyceride reserves and proteins, in contrast to epigean ones. This study thus demonstrates that starvation adaptations are not necessary for cave life, but are rather correlated to the "energetic state" of each ecosystem, and that troglomorphism is not linked to starvation capacities and thus not to the impoverished food availability.

The effects of acute changes in temperature and oxygen availability on cardiac performance in winter flounder (Pseudopleuronectes americanus).

Abstract: Studies on how flatfish cardiovascular function responds to environmental challenges are limited, and have largely relied upon indirect methodologies (i.e. Fick principle). Thus, we measured dorsal aortic blood pressure (P(DA)) and cardiac function in 8 and 15 degrees C-acclimated flounder exposed to graded hypoxia, and in 8 degrees C-acclimated fish exposed to an acute temperature increase to their critical thermal maximum (CTM). The extent of bradycardia in 8 degrees C-acclimated fish (decrease in heart rate of 41%) was consistent with that observed for other teleosts, as was this species' CTM (25.8+/-0.5 degrees C) and its cardiac response to increasing temperature. However, this study provides further examples of how cardiovascular function is controlled differently in the flounder as compared with other fishes. First, the onset of bradycardia in 8 degrees C-acclimated fish occurred earlier than expected for this inactive and hypoxia-tolerant species (60% water air saturation). Second, resting cardiac output was similar in flounder acclimated to 8 and 15 degrees C (approximately 15 mL min(-1) kg(-1)), and hypoxic bradycardia was surprisingly absent at 15 degrees C. Finally, systemic vascular resistance decreased when flounder were exposed to elevated temperature, and this resulted in a 26% fall in P(DA). These are novel findings, however, the extent to which the flounder's behaviour influenced some of the results is unclear.

Fasting suppresses T cell-mediated immunity in female Mongolian gerbils (Meriones unguiculatus).

Abstract: Immune defense is important for organisms' survival and fitness. Small mammals in temperate zone often face seasonal food shortages. Generally fasting can suppress immune function in laboratory rodents and little information is available for wild rodents. The present study tested the hypothesis that Mongolian gerbils (Meriones unguiculatus) could inhibit T cell-mediated immunity to adapt to acute fasting. Forty-two females were divided into the fed and fasted groups, in which the latter was deprived of food for 3 days. After 66 h fasting, half of the gerbils in each group were injected with phosphate buffered saline or phytohaemagglutinin (PHA) solution. T cell-mediated immunity assessed by PHA response was suppressed in the fasted gerbils compared with the fed gerbils. The fasted gerbils had lower body fat mass, wet and dry thymus mass, dry spleen mass, white blood cells, serum leptin and blood glucose concentrations, but higher corticosterone concentrations than those of the controls. Moreover, PHA response was positively correlated with body fat mass and serum leptin levels in the immunochallenged groups. Taken together, acute fasting leads to immunosuppression, which might be caused by low body fat mass and low serum leptin concentrations in female Mongolian gerbils.

Effects of acute changes in salinity and temperature on routine metabolism and nitrogen excretion in gambusia (Gambusia affinis) and zebrafish (Danio rerio).

Abstract: Acute stress may affect metabolism and nitrogen excretion as part of the adaptive response that allows animals to face adverse environmental changes. In the present paper the acute effects of different salinities and temperatures on routine metabolism, spontaneous activity and excretion of ammonia and urea were studied in two freshwater fish: gambusia, Gambusia affinis and zebrafish, Danio rerio , acclimated to 27 °C. The effects on gill morphology were also evaluated. Five salinities (0‰, 10‰, 20‰, 30‰ and 35‰) were tested in gambusia, while four salinities were used in zebrafish (0‰, 10‰, 20‰ and 25‰). Each salinity acute stress was tested alone or in combination with an acute temperature reduction to 20 °C. In gambusia, both salinity and temperature acute stress strongly stimulated urea excretion. Routine oxygen consumption was barely affected by acute salinity or temperature stress, and was reduced by the combined effects of temperature and high salinity. Gills maintained their structural integrity in all stressing conditions; hyperplasia and hypertrophy of mitochondria-rich cells were observed. In zebrafish, temperature and salinity acute changes, both alone and in combination, scarcely affected any parameter tested. The major effect observed was a reduction of nitrogen excretion at 20 °C–25‰; under these extreme conditions a significant structural disruption of gills was observed. These results confirm the high tolerance to acute salinity and temperature stress in gambusia, and demonstrate the involvement of urea excretion modulation in the stress response in this species.

The silk cocoon of the silkworm, Bombyx mori: macro structure and its influence on transmural diffusion of oxygen and water vapor.

Abstract: The cocoon of insect larvae is thought to help conserve water while affording mechanical protection. If the cocoon is a barrier to water loss, then it must also impose a barrier to inward oxygen diffusion. We tested this hypothesis in pupae of the silkworm, Bombyx mori. The rate of water loss and oxygen uptake (VO2) at 25 °C was measured in control pupae in their naturally spun cocoon and in exposed pupae experimentally removed from their cocoon. Additional measurements included the oxygen diffusion coefficient, DO2, of the cocoon wall and dimensions and density of the cocoon fibers. Water loss (as % body mass loss) in both control and exposed pupae was ∼ 1%.day− 1, and was not significantly different between populations. Similarly, VO2 was statistically identical in both control and exposed pupae, at 0.22 ± 0.01 and 0.21 ± 0.02 mL g− 1 · h− 1, respectively. The silk fiber diameter was significantly different in the outer fibers, 26 ± 1 µm, compared with 16 ± 1 µm for the inner fibers lining the cocoon. Inner fibers were also spun significantly more densely (20.8 ± 1.2 mm− 1 transect) than outer fibers (8.3 ± 0.2). Mean DO2 at 25 °C was 0.298 ± 0.002 cm2 · s− 1, approximately the same as unstirred air. These data indicate that the cocoon, while creating a tough barrier offering mechanical protection to the pupa, imposes no barrier to the diffusion of oxygen or water vapor.