Marine Teleost Fish Research Articles

Effect of taurine supplementation on hepatic metabolism and alleviation of cadmium toxicity and bioaccumulation in a marine teleost, red sea bream, Pagrus major.

This study was performed to unravel the mechanism of the beneficial action of taurine on marine teleost fish, red sea bream (Pagrus major), by analyzing the hepatic metabolism. Moreover, the ameliorative effects of the nutrient against cadmium toxicity and bioaccumulation were further evaluated. The fish were fed a diet containing 0% (TAU0%), 0.5% (TAU0.5%), or 5.0% (TAU5.0%) taurine for 40-55days (d) and subjected to cadmium acute toxicity and bioaccumulation tests. Taurine deficiency in feed severely affected growth and the hepatic metabolic profiles of the fish, including a remarkable increase in myo-inositol, aspartate, and ß-alanine in the TAU0% group, which indicates a complementary physiological response to taurine deficiency. For the acute toxicity test, fish were fed the test diets for 55d and were then exposed to different dose of cadmium ranging from 0 to 5.6mg/L for 96h. Fish fed taurine had a higher tolerance to cadmium than those not fed taurine. For the bioaccumulation test, fish were fed the test diets for 40d and then were chronically exposed to 0.2mg/L of cadmium for 28d followed by depuration for 21d. Cadmium concentrations in the liver and muscle of fish fed TAU5.0% were significantly lower than those of fish fed TAU0% for the first 7d of exposure and the first 7d of elimination. Our findings suggest a possible mechanism for the beneficial role played by taurine and that the inclusion of taurine in fish aquaculture feed may reduce cadmium contamination of fish intended for human consumption.

Genetic identification of anisakid nematodes isolated from largehead hairtail (Trichiurus japonicus) in Korea

The nematode species belonging to genus Anisakis occur at their third larval stage in numerous marine teleost fish species worldwide and known to cause accidental human infection through the ingestion of raw or undercooked fish or squids. They may also draw the attention of consumers because of the visual impact of both alive and dead worms. Therefore, the information on their geographical distribution and clear species identification is important for epidemiological survey and further prevention of human infection. For identification of anisakid nematodes species isolated from largehead hairtail (Trichiurus japonicus), polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of internal transcribed spacers of ribosomal DNA were conducted. Mitochondrial cytochrome c oxidase subunit 2 gene was also sequenced, and phylogenetic analysis was conducted. From the largehead hairtail (n = 9), 1259 nematodes were isolated in total. Most of the nematodes were found encapsulated throughout the viscera (56.2 %, 708/1259) or moving freely in the body cavity (41.5 %, 523/1259), and only 0.3 % (4/1259) was found in the muscles. By PCR-RFLP, three different nematode species were identified. Anisakis pegreffii was the most dominantly found (98.7 %, 1243/1259) from the largehead hairtail, occupying 98.7 % (699/708) of the nematodes in the mesenteries and 98.1 % (513/523) in the body cavity. Hybrid genotype (Anisakis simplex × A. pegreffii) occupied 0.5 %, and Hysterothylacium sp. occupied 0.2 % of the nematodes isolated in this study. The largehead hairtail may not significantly contribute accidental human infection of anisakid nematode third stage larvae because most of the nematodes were found from the viscera or body cavity, which are not consumed raw. But, a high prevalence of anisakid nematode larvae in the largehead hairtail is still in concern because they may raise food safety problems to consumers. Immediate evisceration or freezing of fish after catch will be necessary before consumption.

Larval fishes utilize Batesian mimicry as a survival strategy in the plankton

Marine teleost fishes often experience over 99% mortality in the early life stages (eggs and larvae), yet larval survival is essential to population sustainability. Marine fish larvae from a wide range of families display elaborate, delicate features that bear little resemblance to adult forms and hinder their swimming escape ability by increasing drag. Here, we systematically examine the criteria needed for Batesian mimicry to evolve as a survival strategy and present new evidence from in situ imaging technology and simulation modelling to support the hypothesis that many larval morphological features (particularly long, delicate fin rays) and behaviors evolved at least in part through Batesian mimicry of less palatable or noxious gelatinous zooplankton. Many of these or- ganisms (e.g. hydromedusae, cteno phores, and siphon - o phores) are much more abundant than previously recognized. The high predation mortality during the larval phase provides strong potential for selection in favor of maintaining complex and metabolically costly features that mimic gelatinous zooplankton, provided that larger fishes, as selective visual predators, can occasionally be fooled. We conclude that recent ad- vances in our understanding of mimicry combined with information obtained from plankton imaging supports the hypothesis that Batesian mimicry is a widespread survival strategy for larval fishes, which could have broad implications for fish population dy- namics. However, further research is needed in the areas of predator cognition and larval fish behavior in the presence of different predators and models to elu- cidate the circumstan ces in which the larval fish mim- icry hypothesis may apply.

Transcriptome analysis of dormant tomonts of the marine fish ectoparasitic ciliate Cryptocaryon irritansunder lowtemperature.

BackgroundCryptocaryon irritans, a species of obligatory ciliate ectoparasite, can infect various species of marine teleost fish. Cryptocaryon irritans that fall to the seabed or aquarium bottom in winter can form “dormant tomonts” and wake up when the temperature rises the next year. Abundant studies and analyses on the dormant tomonts were carried out at the transcriptome level, in order to investigate the molecular mechanism of C. irritans tomonts entering the dormant state under low-temperature conditions.MethodsThe paired-end sequencing strategy was used to better assemble the entire transcriptome de novo. All clean sequencing reads from each of the three libraries (Group A: untreated blank control; Group B: treated for 24 h at 12 °C; and Group C: developed for 24 h at 25 °C) were respectively mapped back to the transcriptome assembly using the bioinformatics software.ResultsIn this study, 25,695,034, 21,944,467, and 28,722,875 paired-end clean reads were obtained respectively from the three cDNA libraries of the C. irritans tomont by Illumina paired-end sequencing technology. A total of 25,925 unique transcript fragments (unigenes) were assembled, with an average length of 839 bp. Differentially expressed genes (DEGs) were scrutinized; in Group B/A pairwise comparison, 343 genes presented differential expression, including 265 up-regulated genes and 78 down-regulated genes in Group B; in Group C/A pairwise comparison, there were 567 DEGs, including 548 up-regulated genes and 19 down-regulated genes in Group C; and in Group B/C pairwise comparison, 185 genes showed differential expression, including 145 up-regulated genes and 40 down-regulated genes in Group B.ConclusionsThis is the first transcriptomic analytical study of the C. irritans tomonts under low temperature. It can be concluded that most of the genes required for its cell survival under low temperature, or for cell entry into a deeper dormancy state were discovered, and that they might be considered as candidate genes to develop the diagnostic and control measures for cryptocaryoniasis.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1550-1) contains supplementary material, which is available to authorized users.

Distributional patterns of enchodontoid fishes in the Late Cretaceous

Enchodontoidei are extinct marine teleost fishes with a long temporal range and a wide geographic distribution. We propose here to apply Parsimony Analysis of Endemicity (PAE), Track Analysis and Brooks Parsimony Analysis (BPA) to analyze the distributional patterns of these fishes during the Late Cretaceous. Matrices were built according to their respective geological age in the Late Cretaceous. The occurrence data of enchodontoids produced consistent results for the Cenomanian, Turonian, and Santonian. The generalized track found for the Cenomanian (GT 1) can be associated to oceanic currents, whereas for the Turonian the generalized tracks (GT 2, GT 3 and GT 4) were associated with eutrophication/sedimentation effects. During the Santonian, the third Oceanic Anoxic Event (OAE 3) promoted vicariant events that could explain the generalized track found therein (GT 5). The BPA recovered the area formed by Middle East and Europe, which is congruent with GT 1 and GT 5.

Sulfate transporters involved in sulfate secretion in the kidney are localized in the renal proximal tubule II of the elephant fish (Callorhinchus milii).

Most vertebrates, including cartilaginous fishes, maintain their plasma SO4 (2-) concentration ([SO4 (2-)]) within a narrow range of 0.2-1 mM. As seawater has a [SO4 (2-)] about 40 times higher than that of the plasma, SO4 (2-) excretion is the major role of kidneys in marine teleost fishes. It has been suggested that cartilaginous fishes also excrete excess SO4 (2-) via the kidney. However, little is known about the underlying mechanisms for SO4 (2-) transport in cartilaginous fish, largely due to the extraordinarily elaborate four-loop configuration of the nephron, which consists of at least 10 morphologically distinguishable segments. In the present study, we determined cDNA sequences from the kidney of holocephalan elephant fish (Callorhinchus milii) that encoded solute carrier family 26 member 1 (Slc26a1) and member 6 (Slc26a6), which are SO4 (2-) transporters that are expressed in mammalian and teleost kidneys. Elephant fish Slc26a1 (cmSlc26a1) and cmSlc26a6 mRNAs were coexpressed in the proximal II (PII) segment of the nephron, which comprises the second loop in the sinus zone. Functional analyses using Xenopus oocytes and the results of immunohistochemistry revealed that cmSlc26a1 is a basolaterally located electroneutral SO4 (2-) transporter, while cmSlc26a6 is an apically located, electrogenic Cl(-)/SO4 (2-) exchanger. In addition, we found that both cmSlc26a1 and cmSlc26a6 were abundantly expressed in the kidney of embryos; SO4 (2-) was concentrated in a bladder-like structure of elephant fish embryos. Our results demonstrated that the PII segment of the nephron contributes to the secretion of excess SO4 (2-) by the kidney of elephant fish. Possible mechanisms for SO4 (2-) secretion in the PII segment are discussed.

The Rab5A gene of marine fish, large yellow croaker (Larimichthys crocea), and its response to the infection of Cryptocaryon irritans.

Rab GTPases, members of the Ras superfamily, encode monomeric G-proteins. Rab proteins regulate key steps in membrane traffic transport and endocytic pathway of host immune responses. Rab5A is involved in immune regulation, particularly in T cell migration and macrophage endocytosis in higher vertebrates. However, little is known of the molecular structure of Rab5A gene in marine teleost fish species and its expression profile during the parasite infection. In this study, the full-length cDNA sequence and genomic structure of Rab5A gene of the large yellow croaker (Larimichthys crocea) (LycRab5A), one of the most economical marine fishes, were identified and characterized. The LycRab5A protein, containing the ATPase/GTPase binding motifs and the effector molecules binding motifs, was highly homologous to that of other animals. The expression plasmid containing LycRab5A cDNA fused with GST was engineered and transformed into Escherichia coli to produce recombinant protein GST-LycRab5A, which was purified to prepare a polyclonal antibody specifically against LycRab5A. Subcellular localization revealed that LycRab5A expressed in the membrane and cytoplasm. Based on real-time PCR and Western blot analysis, we found that both mRNA and protein of LycRab5A were expressed in all tissues we examined; especially it was highly expressed in blood and gill. Interestingly, both mRNA and protein of LycRab5A were substantially up-regulated when parasitic ciliate protozoan (Cryptocaryon irritans) was infected. The expression of LycRab5A was reached to the maximal level at 24h after infection. The line of evidence suggested that LycRab5A might play an important role in large yellow croaker defense against parasite infection. Moreover, on the basis of protein interaction, it was found that the LycRab5A interacted with myosin light chain (designated as LycMLC), a crucial protein in the process of phagocytosis. This discovery might contribute better understanding to the molecular events involved in fish immune responses.

Adjustments of molecular key components of branchial ion and pH regulation in Atlantic cod (Gadus morhua) in response to ocean acidification and warming

Marine teleost fish sustain compensation of extracellular pH after exposure to hypercapnia by means of efficient ion and acid-base regulation. Elevated rates of ion and acid-base regulation under hypercapnia may be stimulated further by elevated temperature. Here, we characterized the regulation of transepithelial ion transporters (NKCC1, NBC1, SLC26A6, NHE1 and 2) and ATPases (Na(+)/K(+) ATPase and V-type H(+) ATPase) in gills of Atlantic cod (Gadus morhua) after 4 weeks of exposure to ambient and future PCO2 levels (550 μatm, 1200 μatm, 2200 μatm) at optimum (10 °C) and summer maximum temperature (18 °C), respectively. Gene expression of most branchial ion transporters revealed temperature- and dose-dependent responses to elevated PCO2. Transcriptional regulation resulted in stable protein expression at 10 °C, whereas expression of most transport proteins increased at medium PCO2 and 18 °C. mRNA and protein expression of distinct ion transport proteins were closely co-regulated, substantiating cellular functional relationships. Na(+)/K(+) ATPase capacities were PCO2 independent, but increased with acclimation temperature, whereas H(+) ATPase capacities were thermally compensated but decreased at medium PCO2 and 10 °C. When functional capacities of branchial ATPases were compared with mitochondrial F1Fo ATP-synthase strong correlations of F1Fo ATP-synthase and ATPase capacities generally indicate close coordination of branchial aerobic ATP demand and supply. Our data indicate physiological plasticity in the gills of cod to adjust to a warming, acidifying ocean within limits. In light of the interacting and non-linear, dose-dependent effects of both climate factors the role of these mechanisms in shaping resilience under climate change remains to be explored.

Bone-Eating Worms Spread: Insights into Shallow-Water Osedax (Annelida, Siboglinidae) from Antarctic, Subantarctic, and Mediterranean Waters

Osedax, commonly known as bone-eating worms, are unusual marine annelids belonging to Siboglinidae and represent a remarkable example of evolutionary adaptation to a specialized habitat, namely sunken vertebrate bones. Usually, females of these animals live anchored inside bone owing to a ramified root system from an ovisac, and obtain nutrition via symbiosis with Oceanospirillales gamma-proteobacteria. Since their discovery, 26 Osedax operational taxonomic units (OTUs) have been reported from a wide bathymetric range in the Pacific, the North Atlantic, and the Southern Ocean. Using experimentally deployed and naturally occurring bones we report here the presence of Osedax deceptionensis at very shallow-waters in Deception Island (type locality; Antarctica) and at moderate depths near South Georgia Island (Subantarctic). We present molecular evidence in a new phylogenetic analysis based on five concatenated genes (28S rDNA, Histone H3, 18S rDNA, 16S rDNA, and cytochrome c oxidase I–COI–), using Maximum Likelihood and Bayesian inference, supporting the placement of O. deceptionensis as a separate lineage (Clade VI) although its position still remains uncertain. This phylogenetic analysis includes a new unnamed species (O. ‘mediterranea’) recently discovered in the shallow-water Mediterranean Sea belonging to Osedax Clade I. A timeframe of the diversification of Osedax inferred using a Bayesian framework further suggests that Osedax diverged from other siboglinids during the Middle Cretaceous (ca. 108 Ma) and also indicates that the most recent common ancestor of Osedax extant lineages dates to the Late Cretaceous (ca. 74.8 Ma) concomitantly with large marine reptiles and teleost fishes. We also provide a phylogenetic framework that assigns newly-sequenced Osedax endosymbionts of O. deceptionensis and O. ‘mediterranea’ to ribospecies Rs1. Molecular analysis for O. deceptionensis also includes a COI-based haplotype network indicating that individuals from Deception Island and the South Georgia Island (ca. 1,600 km apart) are clearly the same species, confirming the well-developed dispersal capabilities reported in other congeneric taxa. In addition, we include a complete description of living features and morphological characters (including scanning and transmission electron microscopy) of O. deceptionensis, a species originally described from a single mature female, and compare it to information available for other congeneric OTUs.

Heavy metals produce toxicity, oxidative stress and apoptosis in the marine teleost fish SAF-1 cell line

The use of cell lines to test the toxicity of aquatic pollutants is a valuable alternative to fish bioassays. In this study, fibroblast SAF-1 cells from the marine gilthead seabream (Sparus aurata L.) were exposed for 24 h to the heavy metals Cd, Hg, MeHg (Methylmercury), As or Pb and the resulting cytotoxicity was assessed. Neutral red (NR), MTT-tetrazolio (MTT), crystal violet (CV) and lactate dehydrogenase (LDH) viability tests showed that SAF-1 cells exposed to the above heavy metals produced a dose-dependent reduction in the number of viable cells. Methylmercury showed the highest toxicity (EC50 = 0.01 mM) followed by As, Cd, Hg and Pb. NR was the most sensitive method followed by MTT, CV and LDH. SAF-1 cells incubated with each of the heavy metals also exhibited an increase in the production of reactive oxygen species and apoptosis cell death. Moreover, the corresponding gene expression profiles pointed to the induction of the metallothionein protective system, cellular and oxidative stress and apoptosis after heavy metal exposure for 24 h. This report describes and compares tools for evaluating the potential effects of marine contamination using the SAF-1 cell line.

New insights into the factors mediating the onset of puberty in sea bass

In populations of 1-year-old male European sea bass (Dicentrarchus labrax), only large males are able to acquire for the first time a functional competence of their reproductive axis; in other words, to attain puberty. To examine the causes and mechanisms involved in the onset of puberty in this species, a size sorting sampling was carried out to obtain two experimental groups of small and large male fish exhibiting different growth rates. As expected, only large fish reached full spermiogenesis (stage V of testicular development) by the end of the experiment. Our study suggests that fish size is a permissive condition to ensure full effectiveness of the hormonal (Gnrh1, gonadotropins and sexual steroids) actions. Thus, though small fish had endocrine profiles similar to those of large fish, their amplitude was much lower, and was most likely the reason why functional competence of the reproductive axis was not achieved. Moreover, this work provides evidence of the involvement of kisspeptin and Gnrh1 systems in the onset of puberty in a marine teleost fish. It also indicates that very likely kisspeptin and Gnrh1 may regulate gonadotropins and sex steroids at specific stages of testicular development.

Expression of intercellular lipid transport and cholesterol metabolism genes in eggs and early larvae stages of turbot, Scophthalmus maximus, a marine aquaculture species

Growth and energy transfer are critically dependent on effective transport of lipid molecules between tissues and cellular compartments. This process is specific in egg and eleutheroembryos, when energetic and structural lipids, located at the yolk sac, need to be mobilized in order to be incorporated in the new forming embryo, or to produce energy. Here, we describe the transcriptional profile of 11 genes that codify for proteins involved in intercellular lipid transport and cholesterol metabolism during the early development of a marine teleost fish (Scophthalmus maximus), from notochord formation to the period beyond mouth opening. The mRNA expression pattern of genes (apoA1, apoB100, apoE, cetp, mtp, pltp, lipC, lpl, hmgcr1, soat1, lcat) is described and related to previously published lipid levels in larvae and PPARs—peroxisome proliferator-activated receptors—mRNA levels from the same experiment (Cunha et al. in Mar Genomics 10:17–25, 2013). Our findings show that the transcription of genes responsible for apolipoproteins production starts soon before hatching and that activities decline along the development. In contrast, genes responsible for cholesterol synthesis have a low transcription level early in the development and their activity increases later. Apolipoproteins and other genes related to reverse cholesterol transport are possibly under the control of Pparα2, while the expression of extracellular lipid transfer proteins and enzymes involved in cholesterol synthesis is possibly under the simultaneous control of Pparα1 and Pparγ. Generally, the observed transcription of genes involved in lipid transport is in accordance with the lipid composition of the larvae and transcription of master regulators of lipid metabolism such as the nuclear receptors—PPARs.

Structural and functional analysis of ocular regions of five marine teleost fishes (Hippocampus hippocampus, Sardina pilchardus, Gobius niger, Mullus barbatus &Solea solea)

Five marine teleost fishes inhabiting different marine depths namely Hippocampus hippocampus (Linnaeus, 1758), Sardina pilchardus (Walbaum, 1792), Gobius niger (Linnaeus, 1758), Mullus barbatus barbatus (Linnaeus, 1758) and Solea solea (Linnaeus, 1758) were used in the present study. Their retinae and lenses were subjected for histological, scanning electron microscopy, SDS-PAGE and isoenzyme electrophoresis of alkaline phosphatase, malate and glucose-6-phosphate dehydrogenase. The present findings showed variant histological structures with characteristic photoreceptors mainly of either rods for H. hippocampus, M. barbatus and S. solea or cones in S. pilchardus. Mixed photoreceptors are identified in G. niger. The fishes exhibited diversity in protein band expression coincides with change of pattern orientation in lens fibers arrangement and histological structures of retina. Isoenzyme electrophoresis of estimated isoenzyme showed differences between lens and retina of fishes especially H. hippocampus.It can be concluded that the retina and lens of the studied teleost fishes showed apparent varying structure reflecting the isoenzyme characteristic for preserving functional characteristic of vision according to the marine habitat depths.

Bioconcentration and Elimination of the Dithiocarbamate Fungicide Polycarbamate in Marine Teleost Fish and Polychaete.

We evaluated the bioconcentration and elimination of polycarbamate, a popular antifoulant classified as a dithiocarbamate fungicide, in a marine teleost fish, mummichog (Fundulus heteroclitus), and a polychaete (Perinereis nuntia). Following polycarbamate exposure and depuration, the calculated bioconcentration factor (BCF) and elimination rate constant (k2) were 3.1 and 0.17 day(-1) in fish and 1.5-7.8 and 0.13-0.18 day(-1) in the polychaete, indicating that the fungicide has low bioconcentration potential in both organisms. Given the BCF of 3.1, the permissible environmental concentration level of polycarbamate in water was calculated to be 6.1 µg L(-1) to satisfy the threshold pesticide residue levels in fish permitted by law, which is far below the permissible levels in Hiroshima Bay, Japan. Therefore, we consider that the current polycarbamate contamination level in terms of bioconcentration is not likely to be an alarming issue in coastal environments.

Life in the unthinking depths: energetic constraints on encephalization in marine fishes.

Several hypotheses have been proposed to explain the limitation of brain size in vertebrates. Here, we test three hypotheses of brain size evolution using marine teleost fishes: the direct metabolic constraints hypothesis (DMCH), the expensive tissue hypothesis and the temperature-dependent hypothesis. Our analyses indicate that there is a robust positive correlation between encephalization and basal metabolic rate (BMR) that spans the full range of depths occupied by teleosts from the epipelagic (< 200 m), mesopelagic (200-1000 m) and bathypelagic (> 4000 m). Our results disentangle the effects of temperature and metabolic rate on teleost brain size evolution, supporting the DMCH. Our results agree with previous findings that teleost brain size decreases with depth; however, we also recover a negative correlation between trophic level and encephalization within the mesopelagic zone, a result that runs counter to the expectations of the expensive tissue hypothesis. We hypothesize that mesopelagic fishes at lower trophic levels may be investing more in neural tissue related to the detection of small prey items in a low-light environment. We recommend that comparative encephalization studies control for BMR in addition to controlling for body size and phylogeny.

Eye movements are coordinated with pectoral fin beats during locomotion in a marine teleost fish

Animals must simultaneously engage multiple functional systems in order to navigate, feed and survive in complex environments. Nearly all vertebrates perform rapid gaze-shifting eye movements called saccades, but we know little about the behaviour of saccades during rhythmic locomotion. This study examined how saccades are coordinated with locomotor movements in a pectoral-fin-propelled teleost fish, Cymatogaster aggregata, the shiner surfperch. Individual fish were filmed swimming in a flow tank at 10 cm s(-1), and timing data were analysed using circular statistics. The results reveal that C. aggregata generates saccades non-uniformly throughout the pectoral fin cycle. Saccades primarily occur during fin abduction, when a large amount of thrust is produced, and rarely occur during the thrust-free refractory phase. Because vision is known to be impaired during saccades, we hypothesize that C. aggregata synchronizes saccades with periods of high acceleration in order to stabilize retinal images during low-acceleration phases, which are nearly saccade-free.

Ultrastructure and phylogeny of Glugea nagelia sp. n. (Microsporidia: Glugeidae), infecting the intestinal wall of the yellowfin hind, Cephalopholis hemistiktos (Actinopterygii: Serranidae), from the Red Sea.

A new microsporidian species of the genus Glugea Thélohan, 1891 parasitising the marine teleost fish Cephalopholis hemistiktos Rüppell, collected from the Red Sea in Saudi Arabia, is described on the basis of microscopic and molecular procedures. Spherical and whitish xenoma were observed adhering to the intestinal wall. The numerous spores contained within these xenoma, were ovoid to pyriform and measured 4.3-6.0 µm (5.1 µm) in length and 1.8-2.9 µm (2.2 µm) in width. The spore's wall was composed of two thick layers, which were thinner in the area contacting the anchoring disk. The latter appeared at the spore's anterior pole, in an eccentric position to the longitudinal axis. A lamellar polaroplast surrounded the uncoiled portion of the polar filament projected to the basal region of the spore, giving rise to 26-29 turns with winding from the base to the anterior zone of the spore. The posterior vacuole, located at the spore's posterior pole, and surrounded by the polar filament coils, was irregular and composed of light material. Molecular analysis of the rRNA genes, including the ITS region, was performed using maximum parsimony, neighbour-joining and maximum likelihood methods. The ultrastructural features observed, combined with the phylogenetic data analysed, suggest this parasite to be a new species of the genus Glugea. This is the first species of this genus to be reported from Saudi Arabia and is herein named Glugea nagelia sp. n.

Ultrastructure and phylogeny of the parasite Henneguya carolina sp. nov. (Myxozoa), from the marine fish Trachinotus carolinus in Brazil.

Microscopic and molecular procedures are used to describe a new myxosporean species, Henneguya carolina sp. nov., found infecting the intestine of the marine teleost fish Trachinotus carolinus on the southern Atlantic coast of Brazil. Spherical to ellipsoid cysts, measuring up to ~750 µm, display synchronous development. Mature myxospores are ellipsoidal with a bifurcated caudal process. Myxospore body length, width, and thickness are 12.7 ± 0.8 (12.0-13.4) µm, 8.8 ± 0.6 (7.5-9.6) µm, and 5.8 ± 0.4 (5.0-6.4) µm, respectively; 2 equal caudal processes are 16.8 ± 1.1 (15.9-18.0) µm long, and the total myxospore length is 29.4 ± 0.8 (28.4-30.4) µm. Two pyriform polar capsules measure 5.0 ± 0.5 (4.6-5.6) × 2.4 ± 0.4 (1.9-2.9) µm, and each contains a polar filament forming 3 to 4 coils. Sporoplasm is binucleated and presents a spherical vacuole surrounded by numerous globular sporoplasmosomes. Molecular analysis of the small subunit rRNA gene by maximum parsimony, neighbor joining, and maximum likelihood reveals the parasite clustering together with other myxobolids that are histozoic in marine fish of the order Perciformes, thereby strengthening the contention that the host phylogenetic relationships and aquatic environment are the strongest evolutionary signal for myxosporeans of the family Myxobolidae.

Antifreeze protein-induced superheating of ice inside Antarctic notothenioid fishes inhibits melting during summer warming.

Antifreeze proteins (AFPs) of polar marine teleost fishes are widely recognized as an evolutionary innovation of vast adaptive value in that, by adsorbing to and inhibiting the growth of internalized environmental ice crystals, they prevent death by inoculative freezing. Paradoxically, systemic accumulation of AFP-stabilized ice could also be lethal. Whether or how fishes eliminate internal ice is unknown. To investigate if ice inside high-latitude Antarctic notothenioid fishes could melt seasonally, we measured its melting point and obtained a decadal temperature record from a shallow benthic fish habitat in McMurdo Sound, Antarctica. We found that AFP-stabilized ice resists melting at temperatures above the expected equilibrium freezing/melting point (eqFMP), both in vitro and in vivo. Superheated ice was directly observed in notothenioid serum samples and in solutions of purified AFPs, and ice was found to persist inside live fishes at temperatures more than 1 °C above their eqFMP for at least 24 h, and at a lower temperature for at least several days. Field experiments confirmed that superheated ice occurs naturally inside wild fishes. Over the long-term record (1999-2012), seawater temperature surpassed the fish eqFMP in most summers, but never exceeded the highest temperature at which ice persisted inside experimental fishes. Thus, because of the effects of AFP-induced melting inhibition, summer warming may not reliably eliminate internal ice. Our results expose a potentially antagonistic pleiotropic effect of AFPs: beneficial freezing avoidance is accompanied by melting inhibition that may contribute to lifelong accumulation of detrimental internal ice crystals.

Osmoregulatory bicarbonate secretion exploits H(+)-sensitive haemoglobins to autoregulate intestinal O2 delivery in euryhaline teleosts.

Marine teleost fish secrete bicarbonate (HCO3 −) into the intestine to aid osmoregulation and limit Ca2+ uptake by carbonate precipitation. Intestinal HCO3 − secretion is associated with an equimolar transport of protons (H+) into the blood, both being proportional to environmental salinity. We hypothesized that the H+-sensitive haemoglobin (Hb) system of seawater teleosts could be exploited via the Bohr and/or Root effects (reduced Hb-O2 affinity and/or capacity with decreasing pH) to improve O2 delivery to intestinal cells during high metabolic demand associated with osmoregulation. To test this, we characterized H+ equilibria and gas exchange properties of European flounder (Platichthys flesus) haemoglobin and constructed a model incorporating these values, intestinal blood flow rates and arterial–venous acidification at three different environmental salinities (33, 60 and 90). The model suggested red blood cell pH (pHi) during passage through intestinal capillaries could be reduced by 0.14–0.33 units (depending on external salinity) which is sufficient to activate the Bohr effect (Bohr coefficient of −0.63), and perhaps even the Root effect, and enhance tissue O2 delivery by up to 42 % without changing blood flow. In vivo measurements of intestinal venous blood pH were not possible in flounder but were in seawater-acclimated rainbow trout which confirmed a blood acidification of no less than 0.2 units (equivalent to −0.12 for pHi). When using trout-specific values for the model variables, predicted values were consistent with measured in vivo values, further supporting the model. Thus this system is an elegant example of autoregulation: as the need for costly osmoregulatory processes (including HCO3 − secretion) increases at higher environmental salinity, so does the enhancement of O2 delivery to the intestine via a localized acidosis and the Bohr (and possibly Root) effect.