The mussel filter–pump – present understanding,with a re‐examination of gill preparations

Filter feeding in mussels is a secondary adaptation where the gills have become W‐shaped and greatly enlarged, acting as the mussel filter–pump. Water pumping and particle capture in the blue mussel, Mytilus edulis, have been studied over many years. Here, we give a short status of the present understanding of ciliary structure and function of the mussel filter–pump, supplemented with new photo‐microscope and scanning electron microscopy (SEM) pictures of gill preparations. Pumping rate (filtration) and pressure to maintain flow have been extensively studied so the power delivered by the mussel pump to the water flow is known (1.1% of total respiratory power), but the actual cost based on gill respiration is much higher (19%), implying that the cost of maintaining of the large gill pump is considerable and that only relatively little energy can be saved by stopping or reducing the activity of the water‐pumping cilia so that continuous feeding with a ‘minimal scaled’ pump is cheaper than discontinuous feeding with a correspondingly larger pump. According to the present view, the pump proper is the beating lateral cilia (lc) on the gill filaments and particle capture is accomplished by the action of laterofrontal cirri (lfc) transferring particles from the main water current to the frontal gill filament currents driven by frontal cilia (fc). Unexplained aspects include retention efficiency according to particle size and the role of pro‐laterofrontal cilia (p‐lfc) placed between the lfc and fc. The structure of cilia and the mode of ciliary beating have been re‐examined in this study by new high‐resolution light and scanning electron microscopy of isolated gill preparations exposed to serotonin (5‐HT) stimulation which can activate the lc and lfc at low concentrations (10^?6 M), but removes the lfc from the interfilament canals at higher concentrations (10^?5 M).     

Uptake and persistence of human associated Enterococcus in the mussel Mytilus edulis: relevance for faecal pollution source tracking

Aims:  Micro-organisms and molecular markers for microbial source tracking (MST) in coastal waters are often present at low numbers, and often exhibit significant variability in time and space. In this study, we investigated the uptake, accumulation, and persistence of human associated Enterococcus in the mussel Mytilus edulis .
Methods and Results:  The human associated molecular markers esp in Enterococcus faecium , and M66 in Enterococcus faecalis were targetted by PCR in seawater and mussel samples from coastal sites affected by sewage contamination. Both native mussels and mussels transplanted from pristine to polluted sites were included. The results showed that the esp and M66 markers were often not detectable in seawater whereas mussels were enriched in the markers. Human associated E. faecalis accumulated rapidly in M. edulis , and reached maximum levels after 4–6 h with concentration 30–300 times greater than in the surrounding seawater. Enterococcus faecalis retained in M. edulis showed a survival comparable to planktonic E. faecalis in seawater with half lives of 30 and 22 h, respectively. Human associated markers remained detectable for 120 h in M. edulis after faecal contamination.
Conclusions:  The study demonstrated that native and transplanted M. edulis can accumulate and retain human associated molecular markers relevant for MST.
Significance and Impact of the Study:  Mussels should be considered as additional targets in MST studies in coastal waters.     

Enhancement of inward current by serotonin in neurons of Aplysia

In RB cells of Aplysia, serotonin, in the presence of TEA, 4AP and Ba, elicits a voltage-dependent inward current. In Ba-TEA-4AP seawater, RB cells showed a negative slope region (NSR) in their current-voltage (I-V) relationship when measured at the end of 2-s commands from a holding potential of -60 mV. Addition of serotonin to the bathing solution enhanced the NSR. When holding potential was lowered to -10 mV, the NSR as well as the effects of serotonin were greatly reduced. Addition of 20 mM cobalt to the bathing solution blocked both the NSR and the inward current produced by serotonin. Changes in potassium concentration produced no consistent shift in voltage sensitivity nor change in amplitude of the current elicited by serotonin. Intracellular injection of cesium sufficient to broaden action potentials did not block the enhancement of NSR by serotonin. These results support the conclusion that in RB cells, serotonin produces a voltage-dependent current carried by calcium ions.     

Effects of salinity on solute clearance from the freshwater bivalve, Dreissena polymorpha Pallas

Clearance of polyethylene glycol (PEG), inulin, or dextran that had been injected into the hemolymph of the mussel, Dreissena polymorpha, was measured in animals acclimated to pondwater (PW) or 10% seawater (SW). In addition, we measured the clearance of PEG from mussels acutely transferred into 10% SW and following return to PW after acclimation to 10% SW. Clearance values calculated for PW-acclimated mussels ranged from 2.0 to 3.3 ml (g dry tissue ċ h)^-1 and declined to 0.28 ml (g dry tissue ċ h)^-1 in 10% SW-acclimated animals. Transferring mussels into 10% SW resulted in a reduction in PEG clearance from the blood, coincident with the reduction of osmotic gradient. When 10% SW-acclimated mussels were returned to PW the clearance of PEG increased to rates observed in PW-acclimated animals within 1 h. The PEG clearance remained constant during the re-acclimation to PW even though the osmotic gradient declined from about 100 to 30 mosmol kg^-1. Clearance of the solutes used in this study was likely to be a measurement of renal filtration rate. The clearance values appeared to be maximal when the animals were in PW. The limited capacity to increase clearance in the face of an osmotic challenge may be a critical factor in restricting D. polymorpha to freshwater or lower salinity environments with small ranges in salinity.     

Do amiloride and ouabain affect ammonia fluxes in perfused Carcinus gill epithelia?

The effects of inhibitors on the efflux of ammonia (from the basolateral to the apical side, Jb----a) were studied in preparation of isolated Carcinus gills immersed in dilute seawater (DSW) that was identical to the perfusion solution. Adding 10(-4) M amiloride to the solution bathing the gill preparations reduces the efflux of ammonia by 29% relative to the control value. Under experimental conditions, it appears that only about 1% of the amiloride-sensitive influx of Na+ (Ja----b) can be exchanged with NH4+ on an equimolar basis. The ammonium ion is apparently transported at the basolateral side by a carrier-mediated process. Kinetic analyses of the influx of ammonium ions revealed a Km of 36.99 microM and a maximum velocity (Vmax) equal to 19.6 mumol g-1.h-1. Basolaterally applied ouabain (5 x 10(-3) M) and NaCN (10(-3) M) reduced the efflux of ammonia by 46.7 and 42.2%, respectively, suggesting an interaction of NH4+ with the basolaterally located Na+/K+ exchanger in which NH4+ appears to be able to substitute for K+.     

Tissue distribution of a coliphage and Escherichia coli in mussels after contamination and depuration

Experiments were undertaken to determine the tissue distribution of Escherichia coli and a coliphage after contamination of the common mussel (Mytilus edulis). Mussels were contaminated with high levels of feces-associated E. coli and a 22-nm icosahedral coliphage over a 2-day period in a flowing-seawater facility. After contamination, individual tissues were carefully dissected and assayed for E. coli and the coliphage. Contaminated mussels were also analyzed to determine the tissue distribution of the contaminants after 24- and 48-h depuration periods. The majority of each contaminant was located in the digestive tract (94 and 89% of E. coli and coliphage, respectively). Decreasing concentrations were found in the gills and labial palps, foot and muscles, mantle lobes, and hemolymph. Our results indicate that contamination above levels in water occurred only in the digestive tract. Contaminated mussels were depurated in a commercial-scale recirculating UV depuration system over a 48-h period. The percent reductions of E. coli occurred in the following order: digestive tract, hemolymph, foot and muscles, mantle lobes, and gills and labial palps. The percent reductions of the coliphage were different, occurring in the following order: hemolymph, foot and muscles, gills and labial palps, mantle lobes, and digestive tract. Our results clearly demonstrate that E. coli and the coliphage are differentially eliminated from the digestive tract. The two microorganisms are eliminated at similar rates from the remaining tissues. Our results also clearly show that the most significant coliphage retention after depuration for 48 h is in the digestive tract. Thus, conventional depuration practices are inappropriate for efficient virus elimination from mussels.     

Tissue distribution of a coliphage and Escherichia coli in mussels after contamination and depuration.

Experiments were undertaken to determine the tissue distribution of Escherichia coli and a coliphage after contamination of the common mussel (Mytilus edulis). Mussels were contaminated with high levels of feces-associated E. coli and a 22-nm icosahedral coliphage over a 2-day period in a flowing-seawater facility. After contamination, individual tissues were carefully dissected and assayed for E. coli and the coliphage. Contaminated mussels were also analyzed to determine the tissue distribution of the contaminants after 24- and 48-h depuration periods. The majority of each contaminant was located in the digestive tract (94 and 89% of E. coli and coliphage, respectively). Decreasing concentrations were found in the gills and labial palps, foot and muscles, mantle lobes, and hemolymph. Our results indicate that contamination above levels in water occurred only in the digestive tract. Contaminated mussels were depurated in a commercial-scale recirculating UV depuration system over a 48-h period. The percent reductions of E. coli occurred in the following order: digestive tract, hemolymph, foot and muscles, mantle lobes, and gills and labial palps. The percent reductions of the coliphage were different, occurring in the following order: hemolymph, foot and muscles, gills and labial palps, mantle lobes, and digestive tract. Our results clearly demonstrate that E. coli and the coliphage are differentially eliminated from the digestive tract. The two microorganisms are eliminated at similar rates from the remaining tissues. Our results also clearly show that the most significant coliphage retention after depuration for 48 h is in the digestive tract. Thus, conventional depuration practices are inappropriate for efficient virus elimination from mussels.     

鲻和鲮鳃丝的扫描电镜比较观察

对鲻（Mugil cephalus）和鲮（Cirrhina molitorella）的鳃丝表面结构进行了扫描电镜比较观察,结果表明,鲻鳃丝杆状部比鲮粗．鲻鳃小片高度比鲮低;两者鳃丝表面分泌孔洞口径和密度不同;鲻和鲮细胞外被不同,鲻细胞外被稀疏,鲮的则致密复杂;鳃小片细胞和鳃丝表皮细胞的表面形态存在差异,文章还描述了鳃丝表皮形态特异的细胞。     

Pharmacological induction of larval settlement and metamorphosis in the blue mussel Mytilus edulis L

The blue mussel Mytilus edulis L. is an important aquaculture and fouling species in northern seas. Although the general role of chemical cues for settlement of larvae of the blue mussel has been proposed, few studies have focused on induction of settlement and metamorphosis by pharmacological agents. In this study, the induction of larval settlement of the blue mussel by pharmacological compounds was investigated through a series of laboratory experiments with an aim of identifying artificial cues for laboratory bioassay systems in fouling and antifouling research. Gamma-aminobutiric acid (GABA), dihydroxyphenyl L-alanine (DOPA), isobutyl methylxanthine (IBMX) and acetylcholine chloride (ACH) at 10(-7)-10(-2) M as well as KCl at 10-40 mM K+ in excess of the level in normal seawater were tested for their inductive effect on larval settlement. In filtered seawater (FSW) < 9% of the larvae settled after 48 h. Elevated K+ and GABA levels had no effect on larval settlement and metamorphosis. DOPA at 10(-5) M and IBMX at 10(-6)-10(-4) M induced 41-83% larval settlement and ACH at 10(-7)-10(-5) M induced < 40% larval settlement. While the highest settlement rates were observed after 48 h exposure to the chemical, most of the larvae settled within 24 h. Compounds at concentrations of 10(-3)-10(-2) M were either toxic to larvae or retarded the growth of the post-larvae shell. Juveniles resulting from induction by lower concentrations of chemicals had a very high survival rate, completed metamorphosis and grew as well as the juveniles that metamorphosed spontaneously. IBMX at 10(-6)-10(-4) M and L-DOPA at 10(-5) M are effective agents for induction of settlement and metamorphosis for future studies using juvenile M. edulis.     

Gill Na(+)-K(+)-2Cl(-) cotransporter abundance and location in Atlantic salmon: effects of seawater and smolting

Na(+)-K(+)-2Cl(-) cotransporter abundance and location was examined in the gills of Atlantic salmon (Salmo salar) during seawater acclimation and smolting. Western blots revealed three bands centered at 285, 160, and 120 kDa. The Na(+)-K(+)-2Cl(-) cotransporter was colocalized with Na(+)-K(+)-ATPase to chloride cells on both the primary filament and secondary lamellae. Parr acclimated to 30 parts per thousand seawater had increased gill Na(+)-K(+)-2Cl(-) cotransporter abundance, large and numerous Na(+)-K(+)-2Cl(-) cotransporter immunoreactive chloride cells on the primary filament, and reduced numbers on the secondary lamellae. Gill Na(+)-K(+)-2Cl(-) cotransporter levels were low in presmolts (February) and increased 3.3-fold in smolts (May), coincident with elevated seawater tolerance. Cotransporter levels decreased below presmolt values in postsmolts in freshwater (June). The size and number of immunoreactive chloride cells on the primary filament increased threefold during smolting and decreased in postsmolts. Gill Na(+)-K(+)-ATPase activity and Na(+)-K(+)-2Cl(-) cotransporter abundance increased in parallel during both seawater acclimation and smolting. These data indicate a direct role of the Na(+)-K(+)-2Cl(-) cotransporter in salt secretion by gill chloride cells of teleost fish.     

Gills of the medaka (Oryzias latipes): A scanning electron microscopy study

The general morphology and surface ultrastructure of the gills of adult and larvae medaka (Oryzias latipes) were studied in freshwater and seawater using scanning electron microscopy. The gills of all examined fish were structurally similar to those of other teleosts and consisted of four pairs of arches supporting (i) filaments bearing lamellae and (ii) rakers containing taste buds. Three cell types, specifically pavement cells, mitochondria‐rich cells (MRCs), and mucous cells, constituted the surface layer of the gill epithelium. Several distinctive characteristics of medaka gills were noted, including the presence of regularly distributed outgrowth on the lamellae, enlarged filament tips, the absence of microridges in most pavement cells in the filament and lamellae and the presence of MRCs in the arch at the filament base. A rapid mode of development was recorded in the gills of larval fish. At hatching, the larvae already had four arches with rudimentary filaments, rakers, and taste buds. The rudimentary lamellae appeared within 2 days after hatching. These results suggest the early involvement of larval gills in respiratory and osmoregulation activities. The responses of the macrostructures and microstructures of gills to seawater acclimation were similar in larvae and adult fish and included modification of the apical surface of MRCs, confirming the importance of these cells in osmoregulation. The potential roles of these peculiarities of the macrostructures and microstructures of medaka gills in the major functions of this organ, such as respiration and osmoregulation, are discussed.     

The novel correlation of carbonic anhydrase II and anion exchanger 1 in gills of the spotted green pufferfish, Tetraodon nigrovirids

A novel relationship between branchial carbonic anhydrase II (CAII) and anion exchanger 1 (AE1) was investigated in the euryhaline spotted green pufferfish (Tetraodon nigroviridis). The immunoblots revealed that AE1 was only detected in the membrane fraction of gills while CAII can be probed both in the membrane and cytosol fractions of gills. CAII protein abundance in the membrane fraction is salinity dependent. Immunological detection of the membrane fraction CAII protein in gills showed 3.9-fold higher in the hyposmotic (freshwater) group than the hyperosmotic (seawater;35 per thousand) group. In contrast, there was no change in the protein level of cytosolic CAII between seawater and freshwater groups. The whole-mount immunocytochemical staining demonstrated that both AE1 and CAII were colocalized to the Na(+)/K(+)-ATPase-immunoreactive cells in gill epithelium of the pufferfish. The interaction between CAII and AE1 was further identified by co-immunoprecipitation because AE1 was detected in the immunoprecipitates of CAII and vice versa. Our results showed that in pufferfish gills CAII was not only expressed in the cytosol to produce the substrate for AE1 transport during Cl(-) influx but also associated with the plasma membrane via AE1. Obviously, it is essential for the physiological function of AE1 to interact with CAII in the membrane of gill Na(+)/K(+)-ATPase-immunoreactive cells. To our knowledge, this is the first study to demonstrate the interaction of branchial CAII and AE1 in fish. The novel correlation proposed a new model of Cl(-)/HCO(3) (-) transport in gills of the teleosts.     

Studies on the gill ciliation of the American oyster Crassostrea virginica (Gmelin)

Latero-frontal, para-latero-frontal, and frontal ciliary tracts on the gill filaments of Crassostrea virginica (Gmelin) were studied with light microscopy and scanning electron microscopy. Latero-frontal cirri are complex structures composed of varying numbers of paired cilia. The multiple pairs of cilia which constitute a single cirrus are closely appressed for a portion of their length; they then branch laterally from the central axis in a plume-like fashion. Latero-frontal cirri of adjacent gill filaments create a filtration sieve which should be capable of retaining particles smaller than 1 μm in diameter. Para-latero-frontal cilia are short, closely spaced cilia arranged as a staggered row along the frontal side of each tract of latero-frontal cirri. Latero-frontal cirri and para-latero-frontal cilia occur on ordinary, principal, and transitional gill filaments. Frontal ciliary tracts of ordinary filaments are divided into a central, ventrally directed coarse tract, flanked on either side by a dorsally directed fine ciliary tract. The coarse tract is covered by cirri which are comprised of five to eight cilia, while the fine frontal tracts are made up of individually functioning cilia. The frontal ciliary tracts of principal and transitional filaments bear only dorsally directed fine cilia. The unique direction of effective beat of the coarse frontal cirri of ordinary filaments, in combination with the action of fine frontal cilia and the strategic location of mucus producing cells, is used to describe a possible mechanism for the sorting of filtered particles.     

Cytological effects of experimental exposure to Hg on the gill epithelium of the European flat oyster Ostrea edulis: ultrastructural and quantitative changes related to bioaccumulation

The significance of ultrastructural changes in the gill epithelial cells as a parameter of detection of Hg exposure in the flat oyster, Ostrea edulis, was tested by a 34-day exposure to 5 microg l(-1) of Hg. The concentration of Hg (38.76 microg g(-1) dry weight) in gill tissue was maximal after 25 days and then decreased. The histological pattern of gill filaments in control samples did not vary throughout the experiment, except for the volume of mucocytes after 4 days of exposure, as an adaptation to experimental conditions. This volume increased significantly and then decreased according to the accumulation of Hg in the gills. After 18 days, absorptive and ciliated cells of the gill epithelium showed blebs in microvilli membranes, discocilia and swollen mitochondrial cristae. Both cell types showed distinct cellular lysis stages after 25 days of exposure. These are the target cells of Hg and other metals and the reported hypertrophy of mucocytes increase occurs in response to pollution by Hg, which could contribute to the detoxification process.     

The morphology of gills of Haliotis tuberculata (Linnaeus, 1758)

Manganaro, M., Laurà, R., Guerrera, M.C., Lanteri, G., Zaccone, D. and Marino, F. 2011. The morphology of gills of Haliotis tuberculata (Linnaeus, 1758). —Acta Zoologica (Stockholm) 93 : 436–443. Although the morphology of abalone gills has been studied by some authors, up to date no data are available about the gills of Haliotis tuberculata. This study was carried out, by light and electron microscopy, on 10 wild adult H. tuberculata. Gills lamellae produce an undulated surface increasing the area in contact with water. At the level of skeletal rods, we observed a joint‐like structure that allows a checked movement. The left ctenidium is always decidedly larger than the right, probably because of the enormous size of the shell muscle. The cilia permit oxygenated water that leaves the afferent border and is thrust away at the tips of the lamellae by the extremely long cilia. Ciliary movement may take part in sweeping mucous secretions to capture extraneous particles and remove them from the gills. Three types of mucous cells are distributed along the epithelium of the afferent and efferent zones of the gill filament. They seem to play a role in the cleansing of gills in coordination with the muscle contraction and ciliary movement. The presence of microvilli on particular cells reflects their role associated with the absorption of substances from the environment. A haemolymphatic vessel is located in the central zone of the gill filament. The backbone of the haemolymphatic vessel is a chitino‐like structure, which gives support to the gills.     

紫贻贝长期暴露于亚致死剂量的林丹和阿特拉津下的组织学反应

为开发快速、敏感的生物标记物以监测海洋贝类生物中是否存在农药,我们检测了双壳动物紫贻贝长期暴露在亚致死剂量的丙体六氯环乙烷(林丹,γ-HCH)和2-氯4-乙胺基-6异丙胺基-1,3,5-三氮苯(阿特拉津)下的组织学变化。紫贻贝容易积累环境中的杀虫剂,因此,本研究旨在阐明农药的生物积累与组织病理学效应之间的关系。利用GC/MSD分析法定期对贻贝和水样中的林丹与阿特拉津含量进行测定。将贻贝在实验室中培养21天,以使其代谢适应于带有水质控制的封闭式不间断流动系统。随后,30只贝暴露于亚致死剂量的林丹(0.9 mg/L)或阿特拉津(3.583 mg/L)溶液中56天。实验期间,控制重要的参数,比如温度和盐度分别控制在18℃和34‰。在处理28天和56天后取样,检测组织学损坏及吸收的农药量。暴露的紫贻贝每克干重分别能聚集约304.8-372.0μg/g林丹和83.3-137.4μg/g阿特拉津。组织学改变高度集中在鳃的上皮和外套膜组织;上皮与相邻的组织形成分离状态。组织病理学结果显示,抗性机制的激活使紫贻贝能在亚致死压力下存活。组织病理效应范围从浸润反应到以血淋巴细胞出现间质细胞反应为特征。因此,在农药聚集部位的组织学和超微结构的改变是敏感的,并与农药的生物积累具有正相关关系,说明这些改变可能作为农药暴露的生物标记物。     

Anaerobic survival potential of four bivalves from different habitats. A comparative survey

A comparative survey of the anaerobic survival potential of four different bivalve species and the interference of associated bacteria has been carried out. Individuals from both subtidal and intertidal environments were considered by selecting the following species: Mytilus edulis (subtidal epifaunal), Spisula subtruncata (subtidal infaunal), Macoma balthica (intertidal infaunal) and Cerastoderma edule (intertidal infaunal). Anaerobiosis was simulated in the laboratory by subjecting individuals to the following conditions: nitrogen atmosphere, air atmosphere and anoxic seawater incubation. Moreover, the effect of the antibiotic CA (chloramphenicol) was investigated, either as a pre-treatment of individuals kept under normoxic conditions for a week or directly added to the anoxic incubation media. According to survival performances of the individuals, intertidal animals that use to cope with tidal fluctuations in the coastline (emersion processes) had an extraordinary greater capacity to survive aerial exposure as compared to both nitrogen gas and anoxic seawater incubations most likely due to their capacity to perform aerobiosis at certain rate from atmospheric oxygen availability. Specifically, Macoma balthica enlarged its survival potential up to 24.8 days (LT(50)) under air exposure at 12 degrees C as compared to other specific treatments used here (4.9 days). The latter pattern was also observed, although in a much lower magnitude, for the other intertidal species Cerastoderma edule that survived 3.7 and 4.6 days (LT(50)) under nitrogen atmosphere and anoxic seawater incubation, respectively as compared to 9.5 days for emersed individuals. In contrast to the subtidal species, aerial exposure of both intertidal species led to a much higher survival performances than incubation of individuals in anoxic media with the presence of antibiotic. Survival capacity of the subtidal species Mytilus edulis and Spisula subtruncata was statistically similar under air and nitrogen atmospheres and anoxic seawater incubation. Then, subtidal species have a limited ability to air breathing as a conclusion of a similar survival in atmospheric and anoxic seawater incubations. Remarkably, M. edulis represented the only exception when considering longer-term survival capacity compared to the LT(50) values. Indeed, differences in LT(90) values for M. edulis were statistically different, values decreasing significantly from 19.7-19.9 days (under both nitrogen and air atmospheres) to 16.7 days when individuals are incubated in anoxic seawater. This may be due to the adverse effects of anaerobic bacteria that spontaneously proliferate within the static seawater incubations. As well as for S. subtruncata, possible aerobic processes under aerial exposure of mussels seemed to be not significant for the enlargement of its survival potential, since results obtained for both air and nitrogen atmospheres are similar. Pre-treatment with the antibiotic chloramphenicol caused survival capacity to increase by a factor of approx. 2 (M. edulis) and 34-44% (S. subtruncata). In contrast to intertidal species, the direct addition of the antibiotic to the incubation media caused the highest survival performances in both subtidal species. Habitat differences and species-dependent variability must be considered as significant sources of variation when studying the anaerobic performance of individuals using the most common experimental anaerobic techniques to test survival potential.     

Characterization of Na/K-ATPase in Macrobrachium rosenbergii and the effects of changing salinity on enzymatic activity

A ouabain-sensitive Na/K-ATPase kinetic assay system based on the hydrolysis of ATP and the oxidation of NADH was adapted in order to characterize enzymatic activity in gills and examine the effects of changing salinity in Macrobrachium rosenbergii. Maximum inhibition by ouabain occurred at a concentration of 1.4 mM, and the K(m) of the reaction was 0.2 mM. In a first experiment, animals were acclimated to freshwater, 1/3 seawater, 2/3 seawater and full seawater for up to 1 week. Na/K-ATPase activity in front gills was 1. 62+/-0.19 micromol ADP/mg protein per h in freshwater, and was seen to increase slightly in 1/3 seawater (1.88+/-0.19 micromol ADP/mg protein per h) and 2/3 seawater (2.09+/-0.24 micromol ADP/mg protein per h), decreasing slightly in full seawater (1.92+/-0.43 micromol ADP/mg protein per h); however, differences were not significant. Back gills showed slightly higher levels, and a similar pattern of Na/K-ATPase activity. In a second experiment, animals were acclimated to 1/3 seawater and 2/3 seawater, and then transferred to freshwater. However, no changes in activity were seen, indicating that exposure to dilute media did not effect enzymatic activity. Whereas Na/K-ATPase is important in osmoregulatory function in marine euryhaline crustaceans, it may not play a significant role in adaptation in freshwater crustaceans that inhabit a more narrow range of salinities.     

Sodium influx in isolated gills of the freshwater mussel,Ligumia subrostrata

Summary Isolated gills of the freshwater mussel,Ligumia subrostrata, accumulate Na from a pondwater bathing medium. The rate of Na transport by the isolated gill is 13.2±1.1 mol (g dry gill·10 min)^–1 which equals or exceeds the estimated Na transport rate of intact animals. Sodium influx is saturable with aV _max of 13.6±1.2 mol (g dry gill·10 min)^–1 and an affinity (K _s) of 0.17 mM Na/l. The isolated gills survive prolonged exposure to pondwater with a constant of 890 l O₂ (g dry gill·h)^–1 over a 4 h period. Sodium transport in the isolated gills is stimulated 80% above control values by 10^–4 M serotonin, 60% by 0.5 mM cAMP and 60% by 12.5 g/ml nystatin. Sodium influx is inhibited by 0.5 mM amiloride and 1 mM lithium.     

Subcellular distribution of aminotransferases, and pyruvate branch point enzymes in gill tissue from four bivalves

Aspartate aminotransferase (AAT), alanine aminotransferase (ALAT), malic enzyme (ME), malate dehydrogenase (MDH), pyruvate kinase (PK), and phosphoenolpyruvate carboxykinase (PEPCK) activities in cytosolic and mitochondrial fractions of gill tissue from Modiolus demissus (ribbed mussel), Mytilus edulis (sea mussel), Crassostrea virginica (oyster) and Mercenaria mercenaria (quahog) were determined using enzyme assay and starch gel electrophoresis combined with subcellular fractionation. AAT showed distinct mitochondrial and cytosolic isozymes in gills of all these animals. Although ALAT showed distinct mitochondrial and cytosolic isozymes in the gills of oysters, sea mussels and quahogs, only the mitochondrial ALAT was evident in ribbed mussel gill tissue. PK and PEPCK were cytosolic in all these preparations. ME was found only in the mitochondrial fraction of ribbed mussel and quahog gill tissue whereas sea mussel gills showed distinct cytosolic and mitochondrial ME isozymes. With oyster gills, the "cytosolic ME" was electrophoretically identical to the mitochondrial ME indicating that in vivo, the ME is probably mitochondrial. MDH showed distinct cytosolic and mitochondrial isozymes in all bivalve gills tested.