Anterior Gills Research Articles

Comparative studies on the gill rakers of some marine fishes with different feeding habits

The present study aimed to compare between gill rakers of some marine fishes with different feeding habits. A total of 78 specimens in 13 species belonged to 6 fish families: Sparidae (Sparus aurata, Diplodus noct, Rhapdosargus haffara and Boops boops); Family: Mugilidae (Mugil cephalus, Mugil capito and Liza aurata); Family: Siganidae (Siganus rivulatus and Siganus luridus); Family: Synodontidae (Saurida undosquamis and Synodus saurus); Family: Clupeidae (Herklotsichthys quadrimaculatus) and Family: Carangidae (Caranx sexfasciatus) were collected by irregular visits from land fish market in different localities of Egyptian Mediterranean Sea and Suez Gulf; during the period from March, 2014 to November, 2014. Results showed that, the first gill arch formed of one piece, consisting of two limbs (upper and lower limbs). The gill arch carried two rows of gill rakers on its concave border and two rows of gill filaments on its convex one. The gill rakers in the anterior row (oral row) are longer, more in number and more developed than that of the posterior row (aboral row). The gill arch in family Sparidae is bow-like shape. The anterior gill rakers on the first gill arch of most sparid species are short, conical in shape adapted to carnivorous feeding. The gill arch of fish species in Mugilidae is mostly crescent-shaped and rarely bow-shaped. The anterior gill rakers on the first gill arch are long and great in number adapted to detritus feeding. The gill arch in family Synodontidae is V shaped. The anterior gill rakers on the first gill arch appeared as clusters of small tooth patches adapted to piscivores feeding. The gill arch in family Siganidae is hook-like shape. The anterior gill rakers on the first gill arch were needle spine in shape with secondary projections adapted to herbivorous feeding. The gill arch in fish species of Clupeidae is V like shape. Gill rakers are well developed and arranged in only one row. They are numerous and elongated adapted to seizing food items in the plankton feeding. The gill arch in fish species of Carangidae is bow-like shape. Gill rakers are moderate in length and number adapted to carnivorous feeding. According to the different feeding habits, the results showed that, the highest average length of the first gill arch (mm) is recorded in carnivore and piscivore fish. The maximum averages number and the length of the anterior gill rakers are recorded in detritivore fish and plankitivore fish. The highest percentages of the anterior gill raker length /gill arch length ratio (%) are recorded in plankitivore fish and detritivore fish. Results showed that, the maximum inter raker space (µm) and the inter raker space/gill arch length (%) is recorded in carnivore fish. The maximum breadth at the raker base (µm) in the anterior row of rakers is recorded in piscivore fish. The highest breadth at the raker base/gill arch length (%) in the anterior row of rakers is recorded in carnivore, piscivore and planktivore fish. The highest posterior gill raker length (µm) and the posterior gill raker length /gill arch length ratio (%) are recorded in detritivore fish. In conclusion: The morphological characters of the gill rakers in the first gill arch were differ in species studied according to different feeding habits. So that, can be used in determine the fish family and feeding habits.

Molecular characterization of a cDNA encoding Na+/K+/2Cl− cotransporter in the gill of mud crab (Scylla paramamosain) during the molt cycle: Implication of its function in osmoregulation

Although iono-regulatory processes are critical for survival of crustaceans during the molt cycle, the mechanisms involved are still not clear. The Na+/K+/2Cl− cotransporter (NKCC), a SLC12A family protein that transports Na+, K+ and 2Cl− into cells, is essential for cell ionic and osmotic regulation. To better understand the role of NKCC in the molt osmoregulation, we cloned and characterized a NKCC gene from the mud crab, Scylla paramamosain (designated as SpNKCC). The predicted SpNKCC protein is well conserved, and phylogenetic analysis revealed that this protein was clustered with crustacean NKCC. Expression of SpNKCC was detected in all the tissues examined but was highest in the posterior gills. Transmission electron microscopy revealed that posterior gills had a thick type of epithelium for ion regulation while the anterior gills possessed a thin phenotype related to gas exchange. During the molting cycle, hemolymph osmolality and ion concentrations (Na+ and Cl−) increased significantly over the postmolt period, remained stable in the intermolt and premolt stages and then decreased at ecdysis. Meanwhile, the expression of SpNKCC mRNA was significantly elevated (26.7 to 338.8-fold) at the ion re-establishing stages (postmolt) as compared with baseline molt level. This pattern was consistent with the coordinated regulation of Na+/K+-ATPase α-subunit (NKA α), carbonic anhydrase cytoplasmic (CAc) isoform and Na+/H+ exchanger (NHE) genes in the posterior gills. These data suggest that SpNKCC may be important in mediating branchial ion uptake during the molt cycle, especially at the postmolt stages.

Salinity-induced changes in gene expression from anterior and posterior gills of Callinectes sapidus (Crustacea: Portunidae) with implications for crustacean ecological genomics

Decapods represent one of the most ecologically diverse taxonomic groups within crustaceans, making them ideal to study physiological processes like osmoregulation. However, prior studies have failed to consider the entire transcriptomic response of the gill – the primary organ responsible for ion transport – to changing salinity. Moreover, the molecular genetic differences between non-osmoregulatory and osmoregulatory gill types, as well as the hormonal basis of osmoregulation, remain underexplored. Here, we identified and characterized differentially expressed genes (DEGs) via RNA-Seq in anterior (non-osmoregulatory) and posterior (osmoregulatory) gills during high to low salinity transfer in the blue crab Callinectes sapidus, a well-studied model for crustacean osmoregulation. Overall, we confirmed previous expression patterns for individual ion transport genes and identified novel ones with salinity-mediated expression. Notable, novel DEGs among salinities and gill types for C. sapidus included anterior gills having higher expression of structural genes such as actin and cuticle proteins while posterior gills exhibit elevated expression of ion transport and energy-related genes, with the latter likely linked to ion transport. Potential targets among recovered DEGs for hormonal regulation of ion transport between salinities and gill types included neuropeptide Y and a KCTD16-like protein. Using publically available sequence data, constituents for a “core” gill transcriptome among decapods are presented, comprising genes involved in ion transport and energy conversion and consistent with salinity transfer experiments. Lastly, rarefication analyses lead us to recommend a modest number of sequence reads (~10–15M), but with increased biological replication, be utilized in future DEG analyses of crustaceans.

Mechanisms of acid–base regulation in seawater-acclimated green crabs (Carcinus maenas)

The present study investigated acid–base regulatory mechanisms in seawater-acclimated green crabs (Carcinus maenas (L., 1758)). In full-strength seawater, green crabs are osmoconformers so that the majority of the observed responses were attributed to ion fluxes based on acid–base compensatory responses alone. Similar to observations in brackish-water-acclimated C. maenas, seawater-acclimated green crabs exposed to hypercapnia rapidly accumulated HCO3−in their hemolymph, compensating for the respiratory acidosis caused by excess hemolymph pCO2. A full recovery from the decreased hemolymph pH after 48 h, however, was not observed. Gill perfusion experiments on anterior gill No. 5 indicated the involvement of all investigated genes (i.e., bicarbonate transporters, V-(H+)-ATPase, Na+/K+-ATPase, K+-channels, Na+/H+-exchanger, and carbonic anhydrase) in the excretion of acid–base equivalents. The most significant effects were observed when targeting a potentially cytoplasmic and (or) basolaterally localized V-(H+)-ATPase, as well as potentially basolaterally localized bicarbonate transporter (likely a Na+/HCO3−-cotransporter). In both cases, H+accumulated in the hemolymph and CO2excretion across the gill epithelium was significantly reduced or even reversed when blocking bicarbonate transporters. Based on the findings in this study, a working model for acid–base regulatory mechanisms and their link to ammonia excretion in the gill epithelium of C. maenas has been developed.

Making sense of nickel accumulation and sub-lethal toxic effects in saline waters: Fate and effects of nickel in the green crab, Carcinus maenas

In freshwater, invertebrates nickel (Ni) is considered an ionoregulatory toxicant, but its mechanism of toxicity in marine settings, and how this varies with salinity, is poorly understood. This study investigated Ni accumulation and physiological mechanisms of sub-lethal Ni toxicity in the euryhaline green crab Carcinus maenas. Male crabs were exposed to 8.2μg/L (the US EPA chronic criterion concentration for salt waters) of waterborne Ni (radiolabelled with 63Ni) at three different salinities, 20%, 60% and 100% SW for 24h. Whole body Ni accumulation in 20% SW was 3–5 fold greater than in 60% or 100% SW, and >80% of accumulated Ni was in the carapace at all salinities. Ni also accumulated in posterior gill 8, which showed a higher accumulation in 20% SW than in other salinities, a pattern also seen at higher exposure concentrations of Ni (500 and 3000μg/L). Gill perfusion experiments revealed that Ni was taken up by both anterior and posterior gills, but in 20% SW the posterior gill 8, which performs ionoregulatory functions, accumulated more Ni than the anterior gill 5, which primarily has a respiratory function. The sub-lethal consequences of Ni exposure were investigated by placing crabs in Ni concentrations of 8.2, 500, and 3000μg/L at 20, 60 or 100% SW for 24h. In 20% SW, haemolymph Ca levels were significantly decreased by exposure to Ni concentrations of 8.2μg/L or higher, whereas Na concentrations were depressed only at 3000μg/L. Na+/K+-ATPase activity was inhibited at both 500 and 3000μg/L in gill 8, but only in 20% SW. Haemolymph K, Mg, and osmolality were unaffected throughout, though all varied with salinity in the expected fashion. These data suggest that Ni impacts ionoregulatory function in the green crab, in a gill- and salinity-dependent manner.

Serotonin effects in the crab Neohelice granulata: Possible involvement of two types of receptors in peripheral tissues

In crustaceans, serotonin (5-HT) controls various physiological processes, such as hormonal secretion, color changes, reproduction, and metabolism. Since 5-HT injections cause hyperglycemia, this study was designed to further investigate this action of 5-HT in the crab Neohelice granulate, fed with a high-carbohydrate (HC) or a high-protein (HP) diet. The effects of pre-treatment with mammalian 5-HT receptor antagonists, cyproheptadine and methiothepin, were also investigated. A series of in vivo experiments with 3H-5-HT was carried out in order to investigate the presence of putative receptors in peripheral tissues. Since gills were the tissue with the highest labeling in in vivo experiments, in vitro studies with isolated anterior and posterior gills were also conducted. Cyproheptadine blocked the hyperglycemic effect of 5-HT in HP-fed crabs. Methiothepin reduced glycogen levels in the anterior gills of HP crabs and partially blocked the 5-HT-like posture. The injection of 3H-5-HT identified specific binding sites in all the tissues studied and revealed that the binding can be influenced by the type of diet administered to the crabs. Incubation of the anterior and posterior gills with 3H-5-HT and 5-HT confirmed the specificity of the binding sites. Both antagonists inhibited 3H-5-HT binding. In conclusion, this study highlights the importance of serotonin in the control of glucose homeostasis in crustaceans and provides evidences of at least two types of 5-HT binding sites in peripheral tissues. Further studies are necessary to identify the structure of these receptors and their signaling pathways.

Osmoregulation and salinity-induced oxidative stress: is oxidative adaptation determined by gill function?

Osmoregulating decapods such as the Mediterranean green crab Carcinus aestuarii possess two groups of spatially segregated gills: anterior gills serve mainly respiratory purposes, while posterior gills contain osmoregulatory structures. The co-existence of similar tissues serving different functions allows the study of differential adaptation, in terms of free radical metabolism, upon salinity change. Crabs were immersed for 2weeks in seawater (SW, 37 ppt), diluted SW (dSW, 10 ppt) and concentrated SW (cSW, 45 ppt). Exposure to dSW was the most challenging condition, elevating respiration rates of whole animals and free radical formation in hemolymph (assessed fluorometrically using C-H2DFFDA). Further analyses considered anterior and posterior gills separately, and the results showed that posterior gills are the main tissues fueling osmoregulatory-related processes because their respiration rates in dSW were 3.2-fold higher than those of anterior gills, and this was accompanied by an increase in mitochondrial density (citrate synthase activity) and increased levels of reactive oxygen species (ROS) formation (1.4-fold greater, measured through electron paramagnetic resonance). Paradoxically, these posterior gills showed undisturbed caspase 3/7 activity, used here as a marker for apoptosis. This may only be due to the high antioxidant protection that posterior gills benefit from [superoxide dismutase (SOD) in posterior gills was over 6 times higher than in anterior gills]. In conclusion, osmoregulating posterior gills are better adapted to dSW exposure than respiratory anterior gills because they are capable of controlling the deleterious effects of the ROS production resulting from this salinity-induced stress.

Characterization of angiotensin I-converting enzyme from anterior gills of the mangrove crab Ucides cordatus

Angiotensin I-converting enzyme (ACE) is a well-known metallopeptidase that is found in vertebrates, invertebrates and bacteria. We isolated from the anterior gill of the crab Ucides cordatus an isoform of ACE, here named crab-ACE, which presented catalytic properties closely resembling to those of mammalian ACE. The enzyme was purified on Sepharose-lisinopril affinity chromatography to apparent homogeneity and a band of about 72kDa could be visualized after silver staining and Western blotting. Assays performed with fluorescence resonance energy transfer (FRET) selective ACE substrates Abz-FRK(Dnp)P-OH, Abz-SDK(Dnp)P-OH and Abz-LFK(Dnp)-OH, allowed us to verify that crab-ACE has hydrolytic profile very similar to that of the ACE C-domain. In addition, we observed that crab-ACE can hydrolyze the ACE substrates, angiotensin I and bradykinin. The enzyme was strongly inhibited by the specific ACE inhibitor lisinopril (Ki of 1.26nM). However, in contrast to other ACE isoforms, crab-ACE presented a very particular optimum pH, being the substrate Abz-FRK(Dnp)-P-OH hydrolyzed efficiently at pH 9.5. Other interesting characteristic of crab-ACE was that the maximum hydrolytic activity was reached at around 45°C. The description of an ACE isoform in Ucides cordatus is challenging and may contribute to a better understanding of the biochemical function of this enzyme in invertebrates.

Effects of salinity on the accumulation of hemocyte aggregates and bacteria in the gills of Callinectes sapidus, the Atlantic blue crab, injected with Vibrio campbellii

In addition to respiration and ion regulation, crustacean gills accumulate and eliminate injected particles, along with hemocyte aggregates that form in response to those particles. Here we report that the dose of Vibrio campbellii previously shown to induce a decrease in respiration and hemolymph flow across the gill in the Atlantic blue crab, Callinectes sapidus, also triggered the formation of aggregates containing four or more hemocytes in the gills, compared with saline-injected controls. More bacteria were trapped and rendered non-culturable per unit weight by anterior respiratory gills than posterior gills specialized for ion regulation. Further, more bacteria accumulated in the anterior gills of animals held at 30ppt than those at 10ppt. Thus, the role of the gills in immune defense comes at an energetic cost to this and likely to other crustaceans; this cost is influenced by acclimation salinity and the position and specialized function of individual gills.

Review of the Indo-West Pacific ophidiid genera Sirembo and Spottobrotula (Ophidiiformes, Ophidiidae), with description of three new species

Currently, three of the seven described species of Sirembo Bleeker, 1858 and both species of Spottobrotula Cohen & Nielsen, 1978 are considered valid. The present revision is based on 73 specimens of Sirembo known from the Red Sea to Japan and southwards to off northwestern Australia and northern New South Wales and 19 specimens of Spottobrotula known from the Persian Gulf to the Mozambique Channel and from the Andaman Sea. The main characters separating the two genera are the number of long rakers on the anterior gill arch and the form of the sagittal otolith. Based on this distinction Spottobrotula amaculata Cohen & Nielsen, 1982 is transferred to Sirembo. While newly caught specimens can easily be identified by their colour pattern, preserved material requires identification based on meristic and morphometric characters. Three new species are herein described, Sirembo wami, Spottobrotula mossambica and Spottobrotula persica, while Sirembo jerdoni is reported for the first time from Vietnam. These and all other species of the two genera can be identified by combining number of long gill rakers, number of pseudobranchial filaments, number of pectoral fin rays, number and size of scales between dorsal fin origin and lateral line, and prepelvic and pelvic fin length in per cent of SL. The distinction from other genera is discussed.http://zoobank.org/urn:lsid:zoobank.org:pub:5C523099-D826-495E-B880-98B99B1D1714

Uptake and Retention of Microplastics by the Shore Crab Carcinus maenas

Microplastics, plastics particles <5 mm in length, are a widespread pollutant of the marine environment. Oral ingestion of microplastics has been reported for a wide range of marine biota, but uptake into the body by other routes has received less attention. Here, we test the hypothesis that the shore crab (Carcinus maenas) can take up microplastics through inspiration across the gills as well as ingestion of pre-exposed food (common mussel Mytilus edulis). We used fluorescently labeled polystyrene microspheres (8-10 μm) to show that ingested microspheres were retained within the body tissues of the crabs for up to 14 days following ingestion and up to 21 days following inspiration across the gill, with uptake significantly higher into the posterior versus anterior gills. Multiphoton imaging suggested that most microspheres were retained in the foregut after dietary exposure due to adherence to the hairlike setae and were found on the external surface of gills following aqueous exposure. Results were used to construct a simple conceptual model of particle flow for the gills and the gut. These results identify ventilation as a route of uptake of microplastics into a common marine nonfilter feeding species.

Characterization of copper transport in gill cells of a mangrove crab Ucides cordatus

The branchial epithelium of crustaceans is exposed to the environment and is the first site affected by metal pollution. The aim of this work was to characterize copper (Cu) transport using a fluorescent dye, Phen Green, in gill cells of a hypo-hyper-regulator mangrove crab Ucides cordatus. The results showed that added extracellular CuCl2 (0, 0.025, 0.150, 0.275, 0.550 and 1.110μM) showed typical Michaelis–Menten transport for Cu in anterior and posterior gill cells (Vmax for anterior and posterior gills: 0.41±0.12 and 1.76±0.27 intracellular Cu in μM×22.104cells−1×300s−1 respectively and Km values: 0.44±0.04 and 0.32±0.13μM, respectively). Intracellular Cu was significantly higher for posterior gill cells compared to anterior gill cells, suggesting differential accumulation for each gill type. Extracellular Ca at 20mM decreased cellular Cu transport for both anterior and posterior gill cells. Nifedipine and verapamil, calcium channel inhibitors from plasma membrane, decreased Cu transport and affected Km for both gills. These results could be due to a competition between Cu and Ca. Amiloride, a Na/Ca exchanger inhibitor, as well as bafilomycin, a proton pump inhibitor, caused a decrease of intracellular Cu compared to control. Ouabain and KB-R 7943, acting on Na homeostasis, similarly decreased intracellular Cu in both gill cells. Besides that, gill cells exposed to ATP and Cu simultaneously, showed an increase in intracellular copper, which was inhibited by vanadate, an inhibitor of P-type ATPase. These results suggest either the presence of a Cu-ATPase in crab gill cells, responsible for Cu influx, or the effect of a change in electrochemical membrane potential that could also drive Cu to the gill cell interior. Caffeine increased intracellular Cu, suggesting that intracellular Ca could be affecting Cu uptake. Overall the results show that copper uptake in gill cells of crabs is regulated by intracellular Ca, Ca channels and by Na exchangers. This is the first report of Cu transport characterization in whole gill cells of crabs.

Phenotypic flexibility in response to environmental salinity in the euryhaline crab Neohelice granulata from the mudflat and the saltmarsh of a SW coastal lagoon

This study constitutes a first attempt to investigate intraspecific differences in osmoregulatory capacity and digestive and metabolic responses at the biochemical level in relation to hyper- and hypo-regulation in a single species of estuarine crab inhabiting contrasting habitats within a same intertidal area. We compared hemolymph osmolality, key digestive enzymes, glycemia and energy reserves in Neohelice granulata (Dana in Proc Acad Nat Sci Philadelphia 5:247–254, 1851) from the mudflat and saltmarsh of Mar Chiquita coastal lagoon (37°32′/37°45′S-57°19′/57°26′W) under a wide range of salinities (6–60 psu). Individuals from both sites exhibited high and similar osmoregulatory capacity, but while in individuals from mudflat low and high salinities affected lipase activity in hepatopancreas and triglycerides in muscle, in crabs from saltmarsh, high salinities affected glycogen in anterior gills. Low salinity differentially affected free glucose in anterior gills. The results suggest the occurrence of intraspecific distinct digestive and metabolic adjustments in relation to osmoregulatory responses and habitat.

Differential acid–base regulation in various gills of the green crab Carcinus maenas: Effects of elevated environmental pCO2

Euryhaline decapod crustaceans possess an efficient regulation apparatus located in the gill epithelia, providing a high adaptation potential to varying environmental abiotic conditions. Even though many studies focussed on the osmoregulatory capacity of the gills, acid–base regulatory mechanisms have obtained much less attention. In the present study, underlying principles and effects of elevated pCO2 on acid–base regulatory patterns were investigated in the green crab Carcinus maenas acclimated to diluted seawater. In gill perfusion experiments, all investigated gills 4–9 were observed to up-regulate the pH of the hemolymph by 0.1–0.2 units. Anterior gills, especially gill 4, were identified to be most efficient in the equivalent proton excretion rate. Ammonia excretion rates mirrored this pattern among gills, indicating a linkage between both processes. In specimen exposed to elevated pCO2 levels for at least 7days, mimicking a future ocean scenario as predicted until the year 2300, hemolymph K+ and ammonia concentrations were significantly elevated, and an increased ammonia excretion rate was observed. A detailed quantitative gene expression analysis revealed that upon elevated pCO2 exposure, mRNA levels of transcripts hypothesized to be involved in ammonia and acid–base regulation (Rhesus-like protein, membrane-bound carbonic anhydrase, Na+/K+-ATPase) were affected predominantly in the non-osmoregulating anterior gills.

Copper effects on key metabolic enzymes and mitochondrial membrane potential in gills of the estuarine crab Neohelice granulata at different salinities

The estuarine crab Neohelice granulata was exposed (96h) to a sublethal copper concentration under two different physiological conditions (hyperosmoregulating crabs: 2ppt salinity, 1mg Cu/L; isosmotic crabs: 30ppt salinity, 5mg Cu/L). After exposure, gills (anterior and posterior) were dissected and activities of enzymes involved in glycolysis (hexokinase, phosphofructokinase, pyruvate kinase, lactate dehydrogenase), Krebs cycle (citrate synthase), and mitochondrial electron transport chain (cytochrome c oxidase) were analyzed. Membrane potential of mitochondria isolated from anterior and posterior gill cells was also evaluated. In anterior gills of crabs acclimated to 2ppt salinity, copper exposure inhibited hexokinase, phosphofructokinase, pyruvate kinase, and citrate synthase activity, increased lactate dehydrogenase activity, and reduced the mitochondrial membrane potential. In posterior gills, copper inhibited hexokinase and pyruvate kinase activity, and increased citrate synthase activity. In anterior gills of crabs acclimated to 30ppt salinity, copper exposure inhibited phosphofructokinase and citrate synthase activity, and increased hexokinase activity. In posterior gills, copper inhibited phosphofructokinase and pyruvate kinase activity, and increased hexokinase and lactate dehydrogenase activity. Copper did not affect cytochrome c oxidase activity in either anterior or posterior gills of crabs acclimated to 2 and 30ppt salinity. These findings indicate that exposure to a sublethal copper concentration affects the activity of enzymes involved in glycolysis and Krebs cycle, especially in anterior (respiratory) gills of hyperosmoregulating crabs. Changes observed indicate a switch from aerobic to anaerobic metabolism, characterizing a situation of functional hypoxia. In this case, reduced mitochondrial membrane potential would suggest a decrease in ATP production. Although gills of isosmotic crabs were also affected by copper exposure, changes observed suggest no impact in the overall tissue ATP production. Also, findings suggest that copper exposure would stimulate the pentose phosphate pathway to support the antioxidant system requirements. Although N. granulata is very tolerant to copper, acute exposure to this metal can disrupt the energy balance by affecting biochemical systems involved in carbohydrate metabolism.

OSMOLALITY, METALLOTHIONEIN AND NA/KATPASE ACTIVITY IN A MANGROVE CRAB, UCIDES CORDATUS, SUBMITTED TO ACUTE AND CHRONIC COPPER EXPOSURE IN THE WATER

Ucides cordatus were submitted to acute (24h and 96h) and chronic copper (Cu) exposure (15 days), containing 5mg/L CuSO4 in the water. Anterior and posterior gills, hepatopancreas (HP), and aliquots of haemolymph and urine were collected to measure osmolality, metallothionein (MT) concentration and Na/K‐ATPase activity (NKA). Haemolymph showed an increase in osmolality for crabs submitted to 24h Cu exposure, while animals submitted for 96h and 15 days to Cu demonstrated a reduction in osmolality compared to control. There was no change in urine osmolality for animals exposed to Cu for 24h and 15 days, while in the 96h group, there was a small reduction in the osmolality. MT synthesis increased in the anterior gills in crabs exposed to 24h, 96h and 15 days of Cu exposure, while posterior gills showed increased MT concentration in the 24h and 15 days groups, but not in the 96h group, suggesting that there is a regulation between MT concentration and time of exposure to water Cu. There was an increase of NKA in the anterior gills, while in the posterior gills there was reduction in the 24h group and an increase in the 96h and 15 days group compared to controls. For HP, there was an increase of activity for 96h group, while for the 15 days group the values were similar to control. Animals exposed to Cu show a relationship between time of exposure and higher MT concentration and increased NKA in both organs as well as alterations in osmolality.

The microecology of dactylogyrids (Monogenea: Dactylogyridae) on the gills of wild spotted rose snapper Lutjanus guttatus (Lutjanidae) from Mazatlan Bay, Mexico

The spatial distribution and coexistence of monogenean dactylogyrids was assessed on the gills of 63 specimens of wild spotted rose snapper, Lutjanus guttatus (Steindachner), caught in the Mazatlan Bay, Sinaloa, Mexico. Five species are reported: Euryhaliotrema perezponcei Garcia-Vargas, Fajer-Avila et Lamothe-Argumedo, 2008, Euryhaliotrematoides sp., Haliotrematoides spinatus Kritsky et Mendoza-Franco, 2009, H. plectridium Kritsky et Mendoza-Franco, 2009, and H. guttati Garcia-Vargas, Fajer-Avila et Lamothe-Argumedo, 2008. All except E. perezponcei and H. guttati represent new geographical records for the Pacific coast. The most prevalent dactylogyrid species was E. perezponcei (100%), H. plectridium and H. spinatus had > 80% prevalence, and H. guttati and Euryhaliotrematoides sp. had the lowest prevalence. The mean abundance of H. plectridium and E. perezponcei was close to 60 parasites/fish, whereas Euryhaliotrematoides sp. and H. guttati had the lowest abundance. The dactylogyrid species exhibited a tendency for attachment to gill arch 2: 25% attachment occurring on gill arch 1, 30% on 2, 27% on 3 and 18% on 4, and showed a significant preference for the central sector of the gill (42%). Haliotrematoides plectridium had a preference for attachment to gill arches 2 and 3 and the central sector. Haliotrematoides spinatus tended to settle on the gill arches 2 and 3 and had a preference for the central sector. Euryhaliotrema perezponcei tended to settle on the gill arches 1 and 2 and the anterior gill sector. Euryhaliotrematoides sp. and H. guttati did not show a preference for any gill arch or sector. The intraspecific aggregation was stronger than the interspecific aggregation, indicating that all the dactylogyrid species on spotted rose snapper were aggregated, and there was no evidence of competition among the species.

Record of a Rare Ophidiid Fish, Barathrites iris (Actinopterygii: Teleostei: Ophidiiformes), from off Southern Japan

[Two specimens of a rare ophidiid fish, Barathrites iris Zugmayer, 1911, were collected from the abyssal Pacific Ocean, off Kagoshima Prefecture, southern Japan. They agree with the diagnosis of Barathrites in having two pelvic-fin rays, five to six long gill rakers on the anterior gill arch, and a small head (33.5-33.8% of pre-anal-fin length) and eye (1.6-1.8% standard length, SL). Additionally, they can be separated from B. parri Nybelin, 1957, the only other known species of the genus, by their lack of basibranchial tooth patches, smaller eye (10.7-12.3% head length), and shorter pre-dorsal-fin region (17.1-18.9% SL). Previously B. iris was said to differ from B. parri in the number of branchiostegal rays (six versus seven, respectively), but one of the present specimens of B. iris (413 mm SL) has seven branchiostegal rays on the left side and six on the right. Earlier workers have suggested the occurrence of Barathrites in the Pacific Ocean, but no documented records from this ocean appear to exist. This study confirms the occurrence of Barathrites and its type species B. iris in the Pacific Ocean., Two specimens of a rare ophidiid fish, Barathrites iris Zugmayer, 1911, were collected from the abyssal Pacific Ocean, off Kagoshima Prefecture, southern Japan. They agree with the diagnosis of Barathrites in having two pelvic-fin rays, five to six long gill rakers on the anterior gill arch, and a small head (33.5–33.8% of pre-anal-fin length) and eye (1.6–1.8% standard length, SL). Additionally, they can be separated from B. parri Nybelin, 1957, the only other known species of the genus, by their lack of basibranchial tooth patches, smaller eye (10.7–12.3% head length), and shorter pre-dorsal-fin region (17.1–18.9% SL). Previously B. iris was said to differ from B. parri in the number of branchiostegal rays (six versus seven, respectively), but one of the present specimens of B. iris (413 mm SL) has seven branchiostegal rays on the left side and six on the right. Earlier workers have suggested the occurrence of Barathrites in the Pacific Ocean, but no documented records from this ocean appear to exist. This study confirms the occurrence of Barathrites and its type species B. iris in the Pacific Ocean.]

Morphological and Biochemical Variations in the Gills of 12 Aquatic Air-Breathing Anabantoid Fish

All fish species in the Anabantoidei suborder are aquatic air-breathing fish. These species have an accessory air-breathing organ, called the labyrinth organ, in the branchial cavity and can engulf air at the surface of the water to assist in gas exchange. It is therefore necessary to examine the extent of gill modification among anabantoid fish species and the potential trade-offs in their function. The experimental hypothesis that we aimed to test is whether anabantoid fishes have both morphological and functional variations in the gills among different species. We examined the gills of 12 species from three families and nine genera of Anabantoidei. Though the sizes of the fourth gill arch in three species of Trichogaster were reduced significantly, not all anabantoid species had morphological and functional variations in the gills. In these three species, the specific enzyme activity and relative protein abundance of Na(+)/K(+)-ATPase were significantly higher in the anterior gills as compared with the posterior gills and the labyrinth organ. The relative abundance of cytosolic carbonic anhydrase, an indicator of gas exchange, was found to be highest in the labyrinth organ. The phylogenetic distribution of the fourth gill's morphological differentiation suggests that these variations are lineage specific, which may imply a phylogenetic influence on gill morphology in anabantoid species.

Gill ATPase Activities and Microstructures of Juvenile Chinese Mitten Crab, &lt;i&gt;Eriocheir sinensis&lt;/i&gt;, Exposed to Ambient Mercury

Experiments were carried out to investigate the effects of ambient mercury (Hg2+) on gill Na+-K+-ATPase, Ca2+-Mg2+-ATPase activities, and gill microstructure of juvenile Chinese mitten crab, Eriocheir sinensis. Crabs were exposed to ambient 0.00, 0.01, 0.05, 0.10, 0.20 and 0.30 mg/l Hg2+, and sampled on the days of 1, 7, 14 and 30 respectively. The results showed that both ambient Hg2+ concentrations and exposure time had significant effects on both gill Na+-K+-ATPase and Ca2+-Mg2+-ATPase activities of E. sinensis, which showed the tendency to decrease with ambient mercury concentration increasing and exposure time prolonging. While for crabs exposed to 0.01 mg/l Hg2+, no significant difference was observed when compared with the control. Significant interaction between ambient Hg2+ and exposure time on these two ATPases activities was also observed with the lowest values found in crabs exposed to 0.30 mg/l Hg2+ for 30 days. The anterior gill microstructure analysis of E. sinensis exposed to different ambient Hg2+ for 30 days showed that mercury could alter the gill microstructures of E. sinensis. However, there was no significantly structure change appeared in crabs exposed to 0.01 mg/l Hg2+. Combined, these results indicated that though ambient Hg2+ could lead to significant decrease of both gill Na+-K+-ATPase and Ca2+-Mg2+-ATPase activities, and could destroy the gill microstructures, the crab of E. sinensis could acclimate to low Hg2+ contamination less than 0.01mg/l.