Use of a free ocean CO2 enrichment (FOCE) system to evaluate the effects of ocean acidification on the foraging behavior of a deep-sea urchin.

The influence of ocean acidification in deep-sea ecosystems is poorly understood but is expected to be large because of the presumed low tolerance of deep-sea taxa to environmental change. We used a newly developed deep-sea free ocean CO2 enrichment (dp-FOCE) system to evaluate the potential consequences of future ocean acidification on the feeding behavior of a deep-sea echinoid, the sea urchin, Strongylocentrotus fragilis. The dp-FOCE system simulated future ocean acidification inside an experimental enclosure where observations of feeding behavior were performed. We measured the average movement (speed) of urchins as well as the time required (foraging time) for S. fragilis to approach its preferred food (giant kelp) in the dp-FOCE chamber (-0.46 pH units) and a control chamber (ambient pH). Measurements were performed during each of 4 trials (days -2, 2, 24, 27 after CO2 injection) during the month-long period when groups of urchins were continuously exposed to low pH or control conditions. Although urchin speed did not vary significantly in relation to pH or time exposed, foraging time was significantly longer for urchins in the low-pH treatment. This first deep-sea FOCE experiment demonstrated the utility of the FOCE system approach and suggests that the chemosensory behavior of a deep-sea urchin may be impaired by ocean acidification.

Deep-sea faunal communities associated with a lost intermodal shipping container in the Monterey Bay National Marine Sanctuary, CA.

Carrying assorted cargo and covered with paints of varying toxicity, lost intermodal containers may take centuries to degrade on the deep seafloor. In June 2004, scientists from Monterey Bay Aquarium Research Institute (MBARI) discovered a recently lost container during a Remotely Operated Vehicle (ROV) dive on a sediment-covered seabed at 1281 m depth in Monterey Bay National Marine Sanctuary (MBNMS). The site was revisited by ROV in March 2011. Analyses of sediment samples and high-definition video indicate that faunal assemblages on the container's exterior and the seabed within 10 m of the container differed significantly from those up to 500 m. The container surface provides hard substratum for colonization by taxa typically found in rocky habitats. However, some key taxa that dominate rocky areas were absent or rare on the container, perhaps related to its potential toxicity or limited time for colonization and growth. Ecological effects appear to be restricted to the container surface and the benthos within ∼10 m.

Selected regulation of gastrointestinal acid-base secretion and tissue metabolism for the diamondback water snake and Burmese python.

Snakes exhibit an apparent dichotomy in the regulation of gastrointestinal (GI) performance with feeding and fasting; frequently feeding species modestly regulate intestinal function whereas infrequently feeding species rapidly upregulate and downregulate intestinal function with the start and completion of each meal, respectively. The downregulatory response with fasting for infrequently feeding snakes is hypothesized to be a selective attribute that reduces energy expenditure between meals. To ascertain the links between feeding habit, whole-animal metabolism, and GI function and metabolism, we measured preprandial and postprandial metabolic rates and gastric and intestinal acid-base secretion, epithelial conductance and oxygen consumption for the frequently feeding diamondback water snake (Nerodia rhombifer) and the infrequently feeding Burmese python (Python molurus). Independent of body mass, Burmese pythons possess a significantly lower standard metabolic rate and respond to feeding with a much larger metabolic response compared with water snakes. While fasting, pythons cease gastric acid and intestinal base secretion, both of which are stimulated with feeding. In contrast, fasted water snakes secreted gastric acid and intestinal base at rates similar to those of digesting snakes. We observed no difference between fasted and fed individuals for either species in gastric or intestinal transepithelial potential and conductance, with the exception of a significantly greater gastric transepithelial potential for fed pythons at the start of titration. Water snakes experienced no significant change in gastric or intestinal metabolism with feeding. Fed pythons, in contrast, experienced a near-doubling of gastric metabolism and a tripling of intestinal metabolic rate. For fasted individuals, the metabolic rate of the stomach and small intestine was significantly lower for pythons than for water snakes. The fasting downregulation of digestive function for pythons is manifested in a depressed gastric and intestinal metabolism, which selectively serves to reduce basal metabolism and hence promote survival between infrequent meals. By maintaining elevated GI performance between meals, fasted water snakes incur the additional cost of tissue activity, which is expressed in a higher standard metabolic rate.

Gastro-intestinal handling of water and solutes in three species of elasmobranch fish, the white-spotted bamboo shark, Chiloscyllium plagiosum, little skate, Leucoraja erinacea and the clear nose skate Raja eglanteria.

The present study reports aspects of GI tract physiology in the white-spotted bamboo shark, Chiloscyllium plagiosum, little skate, Leucoraja erinacea and the clear nose skate, Raja eglanteria. Plasma and stomach fluid osmolality and solute values were comparable between species, and stomach pH was low in all species (2.2 to 3.4) suggesting these elasmobranchs may maintain a consistently low stomach pH. Intestinal osmolality, pH and ion values were comparable between species, however, some differences in ion values were observed. In particular Ca(2+) (19.67+/-3.65mM) and Mg(2+) (43.99+/-5.11mM) were high in L. erinacea and Mg(2+) was high (130.0+/-39.8mM) in C. palgiosum which may be an indication of drinking. Furthermore, intestinal fluid HCO(3)(-) values were low (8.19+/-2.42 and 8.63+/-1.48mM) in both skates but very high in C. plagiosum (73.3+/-16.3mM) suggesting ingested seawater may be processed by species-specific mechanisms. Urea values from the intestine to the colon dropped precipitously in all species, with the greatest decrease seen in C. plagiosum (426.0+/-8.1 to 0mM). This led to the examination of the molecular expression of both a urea transporter and a Rhesus like ammonia transporter in the intestine, rectal gland and kidney in L. erinacea. Both these transporters were expressed in all tissues; however, expression levels of the Rhesus like ammonia transporter were orders of magnitude higher than the urea transporter in the same tissue. Intestinal flux rates of solutes in L. erinacea were, for the most part, in an inward direction with the notable exception of urea. Colon flux rates of solutes in L. erinacea were all in an outward direction, although absolute rates were considerably lower than the intestine, suggestive of a much tighter epithelia. Results are discussed in the context of the potential role of the GI tract in salt and water, and nitrogen, homeostasis in elasmobranchs.

Effects of salinity on intestinal bicarbonate secretion and compensatory regulation of acid-base balance in Opsanus beta.

Marine teleosts have extracellular fluids less concentrated than their environment, resulting in continual water loss, which is compensated for by drinking, with intestinal water absorption driven by NaCl uptake. Absorption of Cl(-) occurs in part by apical Cl(-)/HCO(3)(-) exchange, with HCO(3)(-) provided by transepithelial transport and/or by carbonic anhydrase-mediated hydration of endogenous epithelial CO(2). Hydration of CO(2) also liberates H(+), which is transported across the basolateral membrane. In this study, gulf toadfish (Opsanus beta) were acclimated to 9, 35 and 50 ppt. Intestinal HCO(3)(-) secretion, water and salt absorption, and the ensuing effects on acid-base balance were examined. Rectal fluid excretion greatly increased with increasing salinity from 0.17+/-0.05 ml kg(-1) h(-1) in 9 ppt to 0.70+/-0.19 ml kg(-1) h(-1) in 35 ppt and 1.46+/-0.22 ml kg(-1) h(-1) in 50 ppt. Rectal fluid composition and excretion rates allowed for estimation of drinking rates, which increased with salinity from 1.38+/-0.30 to 2.60+/-0.92 and 3.82+/-0.58 ml kg(-1) h(-1) in 9, 35 and 50 ppt, respectively. By contrast, the fraction of imbibed water absorbed decreased from 85.9+/-3.8% in 9 ppt to 68.8+/-3.2% in 35 ppt and 61.4+/-1.0% in 50 ppt. Despite large changes in rectal base excretion from 9.3+/-2.7 to 68.2+/-20.4 and 193.2+/-64.9 mumol kg(-1) h(-1) in 9, 35 and 50 ppt, respectively, acute or prolonged exposure to altered salinities was associated with only modest acid-base balance disturbances. Extra-intestinal, presumably branchial, net acid excretion increased with salinity (62.0+/-21.0, 229.7+/-38.5 and 403.1+/-32.9 mumol kg(-1) h(-1) at 9, 35 and 50 ppt, respectively), demonstrating a compensatory response to altered intestinal base secretion associated with osmoregulatory demand.

Postprandial acid-base balance and ion regulation in freshwater and seawater-acclimated European flounder, Platichthys flesus.

The effects of feeding on both acid-base and ion exchange with the environment, and internal acid-base and ion balance, in freshwater and seawater-acclimated flounder were investigated. Following voluntary feeding on a meal of 2.5-5% body mass and subsequent gastric acid secretion, no systemic alkaline tide or respiratory compensation was observed in either group. Ammonia efflux rates more than doubled from 489 +/- 35 and 555 +/- 64 mumol kg(-1) h(-1) under control conditions to 1,228 +/- 127 and 1,300 +/- 154 mumol kg(-1) h(-1) post-feeding in freshwater and seawater-acclimated fish, respectively. Based on predictions of gastric acid secreted during digestion, we calculated net postprandial internal base gains (i.e., HCO (3) (-) secreted from gastric parietal cells into the blood) of 3.4 mmol kg(-1) in seawater and 9.1 mmol kg(-1 )in freshwater-acclimated flounder. However, net fluxes of ammonia, titratable alkalinity, Na(+) and Cl(-) indicated that branchial Cl(-)/HCO (3) (-) and Na(+)/H(+) exchange played minimal roles in counteracting these predicted base gains and cannot explain the absence of alkaline tide. Instead, intestinal Cl(-)/HCO (3) (-) exchange appears to be enhanced after feeding in both freshwater and seawater flounder. This implicates the intestine rather than the gills as a potential route of postprandial base excretion in fish, to compensate for gastric acid secretion.

Temperature and acid-base balance in the American lobster Homarus americanus.

Lobsters (Homarus americanus) in the wild inhabit ocean waters where temperature can vary over a broad range (0-25 degrees C). To examine how environmental thermal variability might affect lobster physiology, we examine the effects of temperature and thermal change on the acid-base status of the lobster hemolymph. Total CO(2), pH, P(CO)2 and HCO(-)(3) were measured in hemolymph sampled from lobsters acclimated to temperature in the laboratory as well as from lobsters acclimated to seasonal temperatures in the wild. Our results demonstrate that the change in hemolymph pH as a function of temperature follows the rule of constant relative alkalinity in lobsters acclimated to temperature over a period of weeks. However, thermal change can alter lobster acid-base status over a time course of minutes. Acute increases in temperature trigger a respiratory compensated metabolic acidosis of the hemolymph. Both the strength and frequency of the lobster heartbeat in vitro are modulated by changes in pH within the physiological range measured in vivo. These observations suggest that changes in acid-base status triggered by thermal variations in the environment might modulate lobster cardiac performance in vivo.

Intestinal anion exchange in teleost water balance.

Simultaneous measurements of all major electrolytes including HCO3(-) and H+ as well as water demonstrated that fluids absorbed by the anterior intestine of the marine gulf toadfish under in vivo-like conditions on an overall net basis are hypertonic at 380 mOsm and acidic ([H+] = 27 mM). This unusual composition of fluids absorbed across the intestinal epithelium is due to the unusual intestinal fluid chemistry resulting from seawater ingestion and selective ion and water absorption along the gastro-intestinal tract. Measurement under near symmetrical conditions with high NaCl concentrations and low MgSO4 concentrations revealed absorption of iso-osmotic and much less acidic fluids by the intestinal epithelium, a situation resembling that of other water absorbing leaky vertebrate epithelia. Reduced luminal NaCl concentrations seen in vivo results in lower absolute water absorption rates but higher Cl-/HCO3(-) exchange rates which are associated with higher net H+ absorption rates. It appears that apical anion exchange is important for net Cl- uptake by the marine teleost intestine especially when luminal NaCl concentrations are low and/or when MgSO4 concentrations are high. Observations indicate that fluid absorption from solutions of low NaCl but high MgSO4 concentrations is energetically more demanding than absorption from NaCl rich solutions at the level of the intestinal epithelium. Furthermore, the high luminal MgSO4 concentration which is an unavoidable consequence of seawater ingestion projects a demand for renal and branchial compensation for intestinal MgSO4 uptake and absorption of hypertonic and acidic fluid by the intestine.

Body fluid volume regulation in elasmobranch fish.

This review addresses an often overlooked aspect of elasmobranch osmoregulation, i.e., control of body fluid volume. More specifically the review addresses the impact of changes in blood volume in elasmobranchs exposed to different environmental salinities. Measurement of blood volume in the European lesser-spotted dogfish, Scyliorhinus canicula, following acute and chronic exposure to 80% and 120% seawater (SW) is reported. In 80%, 100% and 120% SW-adapted S. canicula, blood volume was 6.3+/-0.2, 5.6+/-0.2 and 4.6+/-0.2 mL 100 g(-1) body mass, respectively. Blood volume was significantly higher and lower in 80% and 120% SW-acclimated animals compared to 100% SW controls. Comparisons are made between these results and previously published data. The role of drinking and volume regulation in elasmobranchs is discussed. For the first time measured water reabsorption rates and solute flux rates across the elasmobranch intestinal epithelia are presented. Water reabsorption rates did not differ between 100% SW-adapted bamboo shark, Chiloscyllium plagiosum, and fish acutely transferred to 140% SW. For the most part net solute flux rates and direction for both the 100% and 140% SW groups were the same with the exception of a net efflux of chloride and potassium in the 140% group and influx of these ions in the 100% adapted group. The significance of the intestine as part of the overall elasmobranch osmoregulatory strategy is discussed as is the role of the kidneys, rectal gland and gills in the regulation of body fluid volume in this class of vertebrates.

Feeding and osmoregulation: dual function of the marine teleost intestine.

Experiments on Gulf toadfish Opsanus beta demonstrate how feeding impacts osmoregulation in the marine teleost intestine. A high Ca(2+) diet of pilchards Sardina pilchardus ([Ca(2+)]=404.2 mmol kg(-1)) was compared to a low Ca(2+) diet of common squid Loligo forbesi ([Ca(2+)]=1.3 mmol kg(-1)), as high [Ca(2+)] has been shown to stimulate intestinal anion exchange. Gastrointestinal fluids and blood plasma were collected over a time course from pre-feeding to 216 h post feeding. Following food intake, monovalent ions were largely absorbed across the intestinal epithelium, leaving a fluid rich in divalent ions, which have a lower osmotic coefficient and effectively reduce osmotic pressure in the lumen to allow for enhanced fluid absorption. Concentrations of Cl(-) and HCO (-)(3) in fluid along the gastrointestinal tract of fish fed both diets, particularly 1 and 2 days post-feeding, demonstrate that apical Cl(-)/HCO (-)(3) exchange plays a vital role in postprandial Cl(-) and water absorption. Postprandial acid-base balance disturbance as indicated by plasma alkalinization was limited or absent, indicating compensation for gastric acid secretion in this teleost fish. Plasma osmolality peaked 12 h post-feeding in toadfish fed squid, but was not accompanied by a significant increase in inorganic ion concentrations. Transient fluid secretion by the gastrointestinal tract was evident from reduced luminal Mg(2+) and SO (2-)(4) concentrations for 24-48 h post feeding. Discrepancy between the sum of inorganic osmolytes and measured osmotic pressure was attributed to organic osmolytes, which occurred at high concentrations in the stomach and anterior intestine for up to 24 h post feeding.

Evolutionary aspects of intestinal bicarbonate secretion in fish.

Experiments compared intestinal HCO3- secretion in the intestine of marine teleost Gulf toadfish, Opsanus beta, to representatives of early chondrostean and chondrichthyan fishes, the Siberian sturgeon, Acipenser baerii, and white-spotted bamboo shark, Chiloscyllium plagiosum, respectively. As seen in marine teleosts, luminal HCO3- concentrations were 10-fold plasma levels in all species when exposed to hyperosmotic conditions. While intestinal water absorption left Mg2+ and SO4(2-) concentrated in intestinal fluids up to four-fold ambient seawater concentrations, HCO3- was concentrated up to 50 times ambient levels as a result of intestinal HCO3- secretion. Reduced luminal Cl- concentrations in the intestine of all species suggest that HCO3- secretion also occurs via Cl-/HCO3- exchange in chondrostean and chondrichthyan fishes. Sturgeon began precipitating carbonates from the gut after only 3 days at 14 per thousand, a mechanism utilized by marine teleosts to reduce intestinal fluid osmolality and maintain calcium homeostasis. Analysis of published intestinal fluid composition in the cyclostome Lampetra fluviatilis reveals that this species likely also utilize intestinal HCO3- secretion for osmoregulation. Analysis of existing cyclostome data and our results indicate that intestinal Cl-/HCO3- exchange plays an integral role in maintaining hydromineral balance not only in teleosts, but in all fish (and perhaps other animals) with a need to drink seawater.