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Many Antarctic notothenioid species endemic to the Seasonal Pack-ice Zone have converged on adult blood serum freezing points that are several tenths of a degree above the freezing point of seawater. While these fishes share high adult serum freezing points, the development of their freeze avoidance during ontogeny has not been studied. We investigated this in wild caught juveniles of one such species, Chaenocephalus aceratus (family Channichthyidae), using blood serum antifreeze activity as a proxy for their freeze avoidance. Juvenile serum antifreeze activity was significantly below that of adults through the oldest year 2+ specimens collected. This increased at an estimated rate of 0.368 × 10 ± 0.405 × 10°C day which, if sustained, would leave C. aceratus below their adult serum antifreeze activity levels of 0.57 ± 0.08°C until 4.2 years after hatching. Underlying the 2.7-fold increase in their serum antifreeze activity from late year 0+ juveniles to adults was an even greater 10.4-fold increase in the concentration of their serum antifreeze glycopeptides, which increased proportionally across all of their serum AFGP size isoforms. With insufficient antifreeze activity to avoid freezing in the ice-laden surface waters, both adult and juvenile C. aceratus are most likely restricted to the year round ice-free waters where a metastable supercooled state can be maintained. [ABSTRACT FROM AUTHOR]

Abstract: Nitric oxide (NO)-mediated angiogenesis may play a role in establishing dense retinal vasculatures of Antarctic hemoglobinless icefishes (suborder: Notothenioidei). We hypothesized that loss of hemoglobin (Hb) leads to elevation in [NO] due to decreased degradation of the compound when the NO-scavenger Hb is absent, thereby inducing vascular growth. We found that total mass of NO metabolites, nitrite plus nitrate (NO x ), in plasma is greater in icefishes than in red-blooded notothenioids [e.g. C. aceratus (Hb−), 22.7±2.9μM; N. coriiceps (Hb+), 14.7±1.7μM], suggesting a higher NO load in hemoglobinless animals. High NO levels do not appear to be a result of greater NO synthesis; we consistently measured lower activities of the enzyme catalyzing NO production, nitric oxide synthase, in tissues of icefishes than in Hb-expressing notothenioids [e.g. 96±10 and 216±39 pmol(min g wet wt)−1 in brain tissue of C. aceratus (Hb−) and G. gibberifrons (Hb+), respectively]. Levels of mRNA for hypoxia-induced (HIF-1α and PHD2) and angiogenic genes (VEGF) were similar in red- and white-blooded species, indicating that vascular maintenance in adult animals does not require differences in angiogenic tone. This does not preclude a cause-and-effect relationship between absence of Hb and NO-mediated angiogenesis during earlier ontogenetic stages of icefishes. [Copyright &y& Elsevier]

Abstract: The Trematominae are a particularly interesting subfamily within the antarctic suborder Notothenioidei (Teleostei). The 14 closely related species occupy a large range of ecological of niches, extremely useful for evolutionary and biogeography studies in the Antarctic Ocean. But some Trematomus species can be difficult to identify by using morphological criteria, specially young stages and damaged specimens. Molecular identification would therefore be highly useful, however the suitability of the cytochrome oxidase I gene in a barcoding approach needs to be assessed. We evaluated species delineation within the genus Trematomus comparing morphological identification, nuclear markers (the rhodopsin retrogene and a new nuclear marker pkd1: polycystic kidney disease 1) and COI. We show that Trematomus vicarius is not distinguishable from Trematomus bernacchii with the molecular markers used, and neither is Trematomus loennbergii from Trematomus lepidorhinus. We suggest that until this is investigated further, studies including these species list them as T. loennbergii/T. lepidorhinus group, and keep voucher samples and specimens. Generally, COI gives a congruent result with the rhodopsin retrogene, and except for the previously cited species pairs, COI barcoding is efficient for identification in this group. Moreover pkd1 might not be suitable for a phylogenetic study at this scale for this group. [Copyright &y& Elsevier]

Abstract: Fishes of the perciform suborder Notothenioidei provide an excellent opportunity for studying the evolution and functional importance of evolutionary adaptations to temperature. To understand the unique biochemical features of high-Antarctic notothenioids, it is important to improve our knowledge of these highly cold-adapted stenotherms with new information on their sub-Antarctic relatives. This paper focuses on the oxygen-transport system of two non-Antarctic species, Eleginops maclovinus and Bovichtus diacanthus. Unlike most Antarctic notothenioids, the blood of E. maclovinus and B. diacanthus displays high hemoglobin (Hb) multiplicity. E. maclovinus, the sister group of Antarctic notothenioids, has one cathodal (Hb C) and two anodal components (Hb 1, Hb 2). B. diacanthus, one of the most northern notothenioids, has three major Hbs. The multiple Hbs may have been maintained as a response to temperature differences and fluctuations of temperate waters, much larger than in the Antarctic. Although non-Antarctic notothenioids have never developed cold adaptation, the amino-acid sequence reveals high identity with the globins of Antarctic notothenioids. Hbs of sub-Antarctic notothenioids are characterised by high oxygen affinity and Root effect. Phylogenetic analyses are consistent with the hypothesis that Bovichtidae and Eleginopidae diverged after they became established in more temperate waters north of the Antarctic Polar Front. [Copyright &y& Elsevier]

Nineteen specimens of the rare dragonfish Vomeridens infuscipinnis were evaluated for meristic counts, morphometric measurements, vomerine teeth and the supratemporal canal, anatomical and histological observations of bone, cartilage and lipid, diet, and reproductive status. Seven individuals were measured for buoyancy. All specimens had small vomerine teeth that varied in number. There was also variability in the arrangement of the supratemporal pores and canals. Vomeridens possess a body with little bone and considerable amounts of cartilage and lipid. A mean percentage buoyancy of 1.61% indicated that Vomeridens is nearly neutrally buoyant. Inferences from measurements of buoyancy and from morphological data suggest that Vomeridens lives in an epibenthic water column habitat at 400–900 m. Facilitated by its reduced body density, Vomeridens are likely to forage in the water column by hovering above the substrate. The stomach contents consisted of krill (Euphausia superba), some as large as 46–50 mm.The absolute and relative fecundity in seven female was 1576–2296 oocytes (mean 1889) and 21.3–28.9 oocytes g-1 body weight (mean 25.3), respectively. The reproductive effort in terms of egg diameter, GSI, and absolute and relative fecundity is similar to that for other bathydraconids. [ABSTRACT FROM AUTHOR]

Antarctic fishes survive freezing through the secretion of antifreeze glycoproteins (AFGPs), which bind to ice crystals to inhibit their growth. This mode of action implies that ice crystals must be present internally for AFGPs to function. The entry and internal accumulation of ice is likely to be lethal, however, so how do fishes survive in its presence? We propose a novel function for the interaction between internal ice and AFGPs, namely the promotion of ice uptake by splenic phagocytes. We show here that i) external mucus of Antarctic notothenioids contains AFGPs and thus has a potential protective role against ice entry, ii) AFGPs are distributed widely through the extracellular space ensuring that they are likely to come into immediate contact with ice that penetrates their protective barriers, and iii) using AFGP-coated nanoparticles as a proxy for AFGP adsorbed onto ice, we suggest that internal ice crystals are removed from the circulation through phagocytosis, primarily in the spleen. We argue that intracellular sequestration in the spleen minimizes the risks associated with circulating ice and enables the fish to store the ice until it can be dealt with at a later date, possibly by melting during a seasonal warming event. [ABSTRACT FROM AUTHOR]

Abstract: Notothenioid teleosts underwent major modifications of their genome to adapt to the cooling of the Antarctic environment. In order to identify specific features of the Antarctic teleost immunoglobulin, transcripts encoding the constant region of the IgM heavy chain from 13 Antarctic and non-Antarctic notothenioid species were sequenced. The primary mRNA splicing for the membrane form was found to be atypical in the majority of Antarctic species, because it led to exclusion of two entire constant exons, and to inclusion of 39-nucleotide exons encoding an unusually long Extracellular Membrane-Proximal Domain (EMPD). Genomic DNA analysis revealed that each 39-nucleotide exon fell within a long sequence that was the reverse complement of an upstream region. Deduced amino acid sequence analysis lead to the identification of cysteine encoding codons in the 39-nucleotide exons, but not in the respective sequence counterpart, suggesting that these residues might play an important role in the folding of the EMPD. [Copyright &y& Elsevier]

Since haemoglobins of all animal species have the same haem group, differences in their properties, including oxygen affinity, electrophoretic mobility and pH sensitivity, must result from the interaction of the prosthetic group with specific amino-acid residues in the primary structure. For this reason, fish globins have been the subject of extensive studies in recent years, not only for their structural characteristics, but also because they offer the possibility to investigate the evolutionary history of these ancient molecules in marine and freshwater species living in a great variety of environmental conditions. This review summarizes the current knowledge on the structure, function and phylogeny of haemoglobins of notothenioid fishes. On the basis of crystallographic analysis, the evolution of the Root effect is analysed. Adaptation of the oxygen transport system in notothenioids seems to be based on evolutionary changes, involving levels of biological organization higher than the structure of haemoglobin. These include changes in the rate of haemoglobin synthesis or in regulation by allosteric effectors, which affect the amount of oxygen transported in blood. These factors are thought to be more important for short-term response to environmental challenges than previously believed. [ABSTRACT FROM AUTHOR]

Abstract: Survival of some polar fishes is associated with high levels of circulating antifreeze glycoproteins (AFGPs). AFGP prevent ice growth giving rise to thermal hysteresis. The inhibiting action of AFGPs implies that polar fish contain ice to which AFGPs adsorb. Cryopelagic Pagothenia borchgrevinki, inhabiting the ice-laden waters of McMurdo Sound, Antarctica, were assayed for ice and ice was found on skin, gills, in the intestine, and in the spleen. Two methods used to assess the number of ice crystals in spleens gave comparable results (12.1+/−1.9 and 22+/−3.8 per spleen). Attempts were made to measure the rate of uptake of ice by P. borchgrevinki held in cages immediately beneath the sub-ice platelet layer in McMurdo Sound; uptake was sporadic. Introduction of ice into fish by spray freezing a small patch of the integument resulted in detection of splenic ice after 1h, illustrating that a mechanism exists for ice transport from the periphery to the spleen. Splenic ice did not seem to be eliminated from fish held in ice-free water at −1.6°C for approximately two months. The relatively small number of splenic ice crystals and the slow rate of ice uptake suggest efficient ice barriers exist in P. borchgrevinki. [Copyright &y& Elsevier]

Pogonophryne stewarti, new species, is described from collections made at 1700 m on the continental slope of the Indian Ocean Sector of the Southern Ocean, off Wilkes Land, Antarctica. The new species is assigned to the unspotted P. albipinna species group and is related to P. immaculata. It is distinguished from all other species of the P. albipinna species group by the following combination of characters: a long (about 15-20% SL) mental barbel without a terminal expansion, a shallow head and slender trunk with only slight taper to caudal peduncle, 8-13 middle lateral-line pores, and divergence in the mitochondrial ND2 gene. We discuss the composition and validity of the P. albipinna group, which, up to this point, has been based on three holotype specimens. We provide meristic and morphometric data for seven recently collected adult specimens of P. immaculata. Bayesian analyses of mitochondrial gene sequences for 11 of the now 19 recognized species of Pogonophryne, sampled from all five of the proposed species groups, resulted in reciprocal monophyly of the monotypic species groups. The P. albipinna and P. mentella species groups were each monophyletic and resolved as sister lineages. We provide a revised key for the four species of Pogonophryne classified in the P. albipinna group. [ABSTRACT FROM AUTHOR]

The Trematominae, a subfamily of the Nototheniidae, are typical of the high-Antarctic shelf waters. Within the Trematominae examples of phenotypic plasticity and possible cryptic speciation have been observed. Morphological identification of adult stages can be problematic in cases of high phenotypic plasticity or cryptic speciation. Additionally, postlarval and juvenile stages often have traits still under development and which lack distinction. A microsatellite DNA multiplex of six markers has been developed for Trematomus newnesi (Van Houdt et al. ). This multiplex was tested on five additional trematomid taxa: Pagothenia borchgrevinki, Trematomus bernacchii, Trematomus eulepidotus, Trematomus hansoni and Trematomus scotti. We used these six microsatellite loci to assess the genetic differentiation among species and the resolution power of these loci for individual-based assignment methods. The six species could be well discriminated by conventional methods such as principal component analysis and distance-based methods, and individual Bayesian assignment methods. This marker set can be used for a number of purposes, including the identification of eggs and larval and adult stages. It is also useful for the investigation of recent phylogenetic patterns, as well as the detection of cryptic speciation, which has been suggested for T. bernacchii and T. newnesi but never confirmed with high polymorphic genetic markers. [ABSTRACT FROM AUTHOR]

State III respiration rates were measured in mitochondria isolated from hearts of Antarctic notothenioid fishes that differ in the expression of hemoglobin (Hb) and myoglobin (Mb). Respiration rates were measured at temperatures between 2 and 40°C in Gobionotothen gibberifrons (+Hb/+Mb), Chaenocephalus aceratus (–Hb/–Mb) and Chionodraco rastrospinosus (–Hb/+Mb). Blood osmolarity was measured in all three species and physiological buffers prepared for isolating mitochondria and measuring respiration rates. Respiration rates were higher in mitochondria from G. gibberifrons compared to those from C. aceratus at 2°C, but were similar among all species at temperatures between 10 and 26°C. Respiration rates were significantly lower in icefishes at 35 and 40°C compared to G. gibberifrons. The respiratory control ratio of isolated mitochondria was lower in C. aceratus compared to G. gibberifrons at all temperatures below 35°C. At 35 and 40°C, mitochondria were uncoupled in all species. The Arrhenius break temperature of state III respiration was similar among all three species (30.5 ± 0.9°C) and higher than values previously reported for Antarctic notothenioids, likely due to the higher osmolarity of buffers used in this study. These results suggest that differences in mitochondrial structure, correlated with the expression of oxygen-binding proteins, minimally impact mitochondrial function. [ABSTRACT FROM AUTHOR]

The fish fauna of the Southern Ocean is dominated by species of the perciform suborder Notothenioidei, which constitute 46% of fish species and 90% of biomass. Notothenioids have undergone rapid morphological and ecological diversification and developed physiological adaptations to a cold, highly oxygenated environment. Microbes inhabiting animal intestines include those that perform essential nutritional functions, but notothenioid gut microbial communities have not been investigated using cultivation-independent approaches. We analyzed bacterial 16S rRNA gene sequences obtained from the intestinal tract of Notothenia coriiceps and Chaenocephalus aceratus, which differ in their pelagic distribution and feeding strategies. Both samples showed dominance of Gammaproteobacteria (mostly Vibrionaceae), as has been reported for temperate teleost species. Both samples showed low diversity relative to that reported for other fish microbiota studies, with C. aceratus containing fewer OTUs than N. coriiceps. Despite the small sample size of this preliminary study, our findings suggest that Antarctic notothenioids carry a gut microbiota similar in composition to that of temperate fish, but exhibiting lower species-level diversity. The omnivorous N. coriiceps individual exhibited greater diversity than the exclusively carnivorous C. aceratus individual, which may indicate that increasing herbivory in fish leads to gut microbe diversification, as found in mammals. Lastly, we detected members of taxa containing known microbial pathogens, which have not been previously reported in Antarctic notothenioid fish. [ABSTRACT FROM AUTHOR]

The diversification of the teleost suborder Notothenioidei (Perciformes) in Antarctic waters provides one of the most striking examples of a marine adaptive radiation. Along with a number of adaptations to the cold environment, such as the evolution of antifreeze glycoproteins, notothenioids diversified into eight families and at least 130 species. Here, we investigate the genetic population structure of the humped rockcod ( Gobionotothen gibberifrons), a benthic notothenioid fish. Six populations were sampled at different locations around the Scotia Sea, comprising a large part of the species’ distribution range ( N = 165). Our analyses based on mitochondrial DNA sequence data (352 bp) and eight microsatellite markers reveal a lack of genetic structuring over large geographic distances (ΦST ≤ 0.058, FST ≤ 0.005, P values nonsignificant). In order to test whether this was due to passive larval dispersal, we used GPS-tracked drifter trajectories, which approximate movement of passive surface particles with ocean currents. The drifter data indicate that the Antarctic Circumpolar Current (ACC) connects the sampling locations in one direction only (west–east), and that passive transport is possible within the 4-month larval period of G. gibberifrons. Indeed, when applying the isolation-with-migration model in IMA, strong unidirectional west-east migration rates are detected in the humped rockcod. This leads us to conclude that, in G. gibberifrons, genetic differentiation is prevented by gene flow via larval dispersal with the ACC. [ABSTRACT FROM AUTHOR]

The article discusses the potential for new model organisms to emerge within the research realm of developmental biology. Clinical research has traditionally been conducted on a few specific species, including the mouse (Mus musculus), the fruitfly (Drosophila melanogaster), and the zebrafish (Danio rerio). There are inherent difficulties in straying from these established models, including the need for new genetic and genomic tools. Nonetheless, other organisms such as Notothenioidei (ice fish) are being considered as model organisms. Among this group of fish are the three-spined stickleback (Gasterosteus aculeatus) and the Mexican cave fish (Astyanax mexicanus).

Parental care and nest guarding behaviors have been described for all major clades of Antarctic notothenioid fishes. In possible association with this reproductive behavior, there is no evidence of significant fish egg predation in notothenioids so far. Here, we report a recently ingested fish egg mass in the stomach of a large specimen of the voracious bullhead notothen Notothenia coriiceps caught in Paradise Bay, West Antarctic Peninsula, in mid-January 2019. Using molecular markers, the preyed eggs were identified as being emerald notothens Trematomus bernacchii , which enabled us to discuss aspects of the emerald notothen reproductive ecology. Our estimation of 4443 ingested eggs of 3.94 mm in average diameter, coupled with comparable literature data, suggested that this bullhead notothen ate the entire spawn. Based on an estimated age of the T. bernacchii embryos at 30 to 45 days post-fertilization, the spawning event likely occurred in December. Although it is known from a close location to our study site that emerald notothens use sponge cavities as spawning substrates with females guarding nests, we also recovered a small fragment of algae in the stomach content of the bullhead notothen, suggesting that the egg mass could have also been laid on algae. Our findings suggests that in this case of predator-prey interaction between two sympatric notothenioid species, there was a lack of effectiveness in egg protection, revealing a possible limit of egg guarding in the Antarctic ichthyofauna. [Display omitted] • First report on the predation of fish eggs clutch in Antarctic notothenioids. • Molecular techniques solve challenging prey species identification in diet studies. • Parental egg guarding strategy in notothenioid fishes shows potential limitations. • Inshore sites serve as spawning grounds for Antarctic notothenioids. [ABSTRACT FROM AUTHOR]

Abstract: This mini-review makes a survey and a summary of some major issues concerning the chromosomal organisation of ribosomal genes in fish genomes, by using Notothenioidei as the model. The increasing body of information, published during the last two decades on the chromosomal mapping of the two ribosomal genes classes (45S rDNA and 5S rDNA) in notothenioids, makes it possible to recognise the main evolutionary trends across the phylogeny of the group. As one of the major features, the rDNA clusters are organised in a single chromosomal locus in most of the species. This locus is located at different positions along the chromosomes in the basal groups (non-Antarctic Clade), whereas it maintains a strongly conserved location in the cold-adapted species (Antarctic Clade). Important structural changes, leading to the co-localisation of the two ribosomal gene classes, occurred early in the notothenioid phylogeny, perhaps in the common ancestor of the Eleginopidae and Nototheniidae. The cytogenetic evidences indicate that an increased amount of ribosomal genes, organised in two large chromosomal loci, is present in the giant Antarctic fish Dissostichus mawsoni. This gain in rRNA genes is an important genomic change, having possible implications for the fitness of this notothenioid fish that combines large size, pelagic lifestyle and cold-adaptation. [Copyright &y& Elsevier]

The article presents a discussion on the intriguing phenomena called "Icefish Paradox," which has been a curiosity for biochemists, regarding the task of neuroglobin in Antarctic hemoglobin-less icefish. The Southern Ocean area of Antarctica has made organisms like fishes to resort to adaptations to counter the cold. One such adaptation and modification is seen among the Antarctic fish suborder Notothenioidei, which displays modification of like decreased amounts and multiplicity of hemoglobins.

A primary objective of the ICEFISH 2004 cruise was to collect and study notothenioid fishes from remote localities in the Atlantic sector of the Southern Ocean. Nearly 1 month was devoted to bottom trawling for fishes on the shelf and upper slope (to 1,000 m) areas around Shag Rocks, South Georgia, South Sandwich Islands and Bouvetøya. The focus was on the latter two locations, because their faunas are more poorly known. Eight species were collected at Shag Rocks with Patagonotothen guntheri most abundant; 17 at South Georgia with Lepidonotothen nudifrons, L. larseni and Gobionotothen gibberifrons most abundant; 13 at the South Sandwich Islands with L. larseni, L. nudifrons and G. gibberifrons most abundant; and 11 at Bouvetøya with L. larseni, Macrourus holotrachys and L. squamifrons most abundant. Ten new locality records were established: Shag Rocks (1), South Georgia (1), South Sandwich Islands (5), South Sandwich Trench at 5,350 m (1) and Bouvetøya (2). Total known demersal fish diversity on the shelf and upper slope at Shag Rocks/South Georgia, South Sandwich Islands and Bouvetøya is 42, 31 and 17 species, respectively. To examine population structure in the four most abundant notothenioids at Bouvetøya ( L. larseni, L. squamifrons, Notothenia coriiceps and Chaenocephalus aceratus), we examined the ND2 portion of mitochondrial DNA. Chaenocephalus aceratus, N. coriiceps and L. larseni exhibited no significant genetic differentiation in comparison with samples from localities in the Scotia Sea and the Antarctic Peninsula. However, L. squamifrons showed significant genetic differentiation between the South Shetlands and Bouvetøya populations ( F ST = 0.189, P = 0.015). Thus, these data combined with previous studies of two other notothenioids suggest that five of the six notothenioid species at Bouvetøya are not genetically differentiated from other localities in the Atlantic sector of the Southern Ocean. The location of Bouvetøya within the Antarctic Circumpolar Current and the long (1–2 years) pelagic stages of the notothenioids at Bouvetøya may be at least partly responsible for this genetic homogeneity. [ABSTRACT FROM AUTHOR]

We evaluated the condition factor (K), an index of weight per unit length, in a sample of 49 761 longline-caught specimens of Antarctic toothfish, Dissostichus mawsoni, from the Ross Sea. Our sample consisted of specimens from northern (60-70°S) and southern (70-78°S) regions. Fifty percent of the northern sample and 4.9% of the southern sample were large individuals with a low condition factor (Kah ≤ 1.0182) and displaying an "axe handle (ah)" morphology with a notably thinner trunk. In the northern sample 55.4% of the males and 43.2% of the females were less than Kah. In the southern region 5.3% of the males and 4.5% of the females were less than Kah. The axe handle morphology is attributable to the metabolic loss of muscular and subcutaneous lipid stores, and probably proteins, from white muscle. Our discussion considers energy metabolism as related to migration, feeding and reproduction. We suggest that migration, a life history trait of the phyletically basal notothenioid clades Pseudaphritis and Eleginops, persists in D. mawsoni. The spawning migration from the southern shelf to ridges and seamounts in the north may reflect fidelity to historic spawning grounds. As evidenced by the axe handle specimens neutral buoyancy of large D. mawsoni may be ephemeral, fluctuating over the course of adult life. [ABSTRACT FROM AUTHOR]

A new species of notothenioid fish, Pogonophryne bellingshausenensis n. sp., is described from the Bellingshausen Sea, Antarctica. The new species belongs to the dorsally-spotted “mentella” group of the genus and is characterized by having a short (about 13% SL) mental barbel with a short (about 16% of barbel length), narrow (barely wider than the stalk), and relatively inconspicuous terminal expansion composed of short, irregular, fingerlike processes. Compared to most other dorsally-spotted species of Pogonophryne (“barsukovi”, “marmorata”, and “mentella” groups), P. bellingshausenensis has a relatively wide (about 7% SL) interorbital region. An unspotted patch on the median dorsal surface of the head, posterior to the posttemporal ridges and anterior to the first dorsal fin, has not been observed previously in any dorsally-spotted species. The holotype was collected at 1,947 m, one of the deepest records for any species of Pogonophryne. A revised key to the ten species of the “mentella” group of Pogonophryne is also provided. [ABSTRACT FROM AUTHOR]

We analysed cytochrome oxidase I (COl) barcodes for 35 putative fish species collected in the Scotia Sea, and compared the resultant molecular data with field-based morphological identifications, and additional sequence data obtained from GenBank and the Barcode of Life Data System (BOLD). There was high congruence between morphological and molecular classification, and COl provided effective species-level discrimination for nearly all putative species. No effect of geographic sampling was observed for COl sequence variation. For two families, including the Liparidae and Zoarcidae, for which morphological field identification was unable to resolve taxonomy, DNA barcoding revealed significant species-level divergence. However, the dataset lacked sufficient sensitivity for resolving species within the Bathydraco and Artedidraco genera. Analysis of cytochrome b for these two genera also failed to resolve taxonomic identity. The data are discussed in relation to emergent priorities for additional taxonomic studies. We emphasize the utility of DNA barcoding in providing a valuable taxonomic framework for fundamental population studies through assigning life history stages or other morphologically ambiguous samples to parental species. [ABSTRACT FROM AUTHOR]

Abstract: Notothenioids represent an adaptive radiation of teleost fishes in the frigid and ice-laden waters of the Southern Ocean surrounding Antarctica. Phylogenetic hypotheses for this clade have resulted primarily from analyses of mtDNA gene sequences, and studies utilizing nuclear gene DNA sequence data have focused on particular sub-clades of notothenioid fishes. In this study, we provide the first phylogenetic analysis of notothenioids using both mtDNA and nuclear gene sequences for a comprehensive sampling of all major lineages in the clade. Maximum parsimony and Bayesian analyses of aligned mtDNA genes, an aligned nuclear gene (S7 ribosomal protein intron 1), and combined dataset containing the mtDNA and nuclear genes resulted in phylogenies that contained the previously identified Antarctic and High Antarctic Clades. There were areas of agreement and disagreement between different datasets and methods of phylogenetic analysis, and the phylogenies resulting from the nuclear encoded S7 ribosomal protein intron 1 sequences were considerably less resolved than those inferred from mtDNA gene sequences. However, we anticipate increased resolution of the notothenioid phylogeny from future analyses that sample DNA sequences from several nuclear genes. [Copyright &y& Elsevier]

The Falkland’s mullet, Eleginops maclovinus, is the only modern representative of the Sub-Antarctic family Eleginopidae, suborder Notothenioidei. Based on specimens from the Falkland Islands/Islas Malvinas, the Magellan Straits, and the southern coast of Chile, we have established the specific karyotype by conventional cytogenetic methods and have mapped the chromosomal loci of the ribosomal genes by fluorescence in situ hybridization (FISH). With respect to the basal notothenioid family Bovichtidae and to the hypothetical basal condition of the suborder (diploid number = 48, fundamental number = 48), E. maclovinus displays a slightly derived karyotype (diploid number = 48, fundamental number = 54). In contrast to the bovichtids, the 45S and 5S ribosomal DNAs are co-localized to a single chromosome pair. Condensation of the ribosomal genes to a single locus is likely to represent an intermediate stage in the evolution of notothenioid karyology. Features unique to E. maclovinus (e.g., morphology of its large, rDNA-bearing chromosome pair) probably result from divergence during the long evolutionary isolation of the family. [ABSTRACT FROM AUTHOR]

Chaenocephalus aceratus is one of the most abundant Antarctic icefish species in the Atlantic sector and has been a by-catch species in the fishery for mackerel icefish, Champsocephalus gunnari, between the mid-1970s and mid-1980s at South Georgia, South Orkney, and South Shetland Islands. The species became the target of the fishery in particular seasons, such as at South Georgia in 1977/78. In our paper, we report results on genetic differentiation for 11 microsatellite loci in C. aceratus samples collected at the South Shetlands and Elephant Island. This study represents the first report on microsatellite variability of an icefish species. Our results support the evidence from previous studies on differences in infestation patterns of parasites that a single panmictic population of C. aceratus exists, spanning the two sampling sites separated by about 100 km. Moreover, our study indicates the presence of a significant genetic differentiation between individual year-classes pointing out the existence of dynamic processes acting at the population genetic level, according to recent results for broadly distributed marine species. Both small effective population size and immigration from unsampled differentiated stocks may be at the base of the differentiation found in C. aceratus. [ABSTRACT FROM AUTHOR]

Mitochondrial gene content shows extensive variation among eukaryotes, but is remarkably compact and static in bilateral animals. Mitochondrial genomes of bilaterians typically contain two rRNA, 22 tRNA, and 13 protein-coding genes. In this study, we report that the mitochondrial genomes of Antarctic fishes of the suborder Notothenioidei (Perciformes) lack two adjacent genes encoding NADH 6 dehydrogenase (ND6) and tRNAGlu. Loss of the ND6 gene is reported for the first time in an animal mitochondrial genome, and is considered an extremely rare evolutionary event. Dot blot and ND6 transcript detection analyses found no evidence of mitochondrial ND6 gene copies in heteroplasmy or of a functional ND6 gene copy in the nuclear genome, respectively. Hence, we concluded that ND6 function was lost in Antarctic notothenioids, and could be compensated for by functional changes in other proteins of the mitochondrial respiratory system. [ABSTRACT FROM AUTHOR]

The complete mitochondrial genome of Notothenia rossii was obtained using PacBio Sequel long-read sequencing platform. The mitogenome of N. rossii was circular form and 18,274 bp long, which consists of 13 protein-coding genes, 24 tRNAs, 2 rRNAs, and non-coding control region. Particularly, we found duplicated tRNAThr and tRNAPro in addition to the typical 22 tRNAs. The phylogenetic tree revealed that N. rossii was most closely related to N. coriiceps among species in the Nototheniidae clade within the suborder Notothenioidei. [ABSTRACT FROM AUTHOR]

Abstract: Given the unique thermal history of the Antarctic continent, fishes of dominant suborder Notothenioidei offer a remarkable opportunity to study the physiological and biochemical characters gained and, conversely, lost during their evolutionary history and to map this information on the species phylogenetic trees. The availability of phylogenetically related notothenioid taxa living in a wide range of latitudes (in the Antarctic, sub-Antarctic and temperate regions) allows to look into the molecular bases of environmentally driven gene birth and death. This evolutionary perspective has also been supported by comparison of some features of the hemoprotein devoted to the oxygen transport in fish species living in the other polar region, the Arctic. The Arctic and Antarctic marine ichthyofaunas differ by age and isolation. Fishes of the two polar regions have undergone different regional histories which have engineered the physiological diversities, so that Antarctic fish are much more stenothermal than Arctic ones. Understanding the mechanisms of phenotypic response to cold exposure in species living at different latitudes in polar habitats offers fundamental insights into environmental adaptations. This review aims at surveying the current knowledge of molecular structure, functional features, phylogeny and adaptations of the hemoglobin of fish thriving in the Antarctic, sub-Antarctic and Arctic regions (with some excursions in the temperate latitudes). Investigating the evolutionary adaptations of hemoglobins to these environments can provide new insights into adaptation currently studied merely in temperate organisms, and can shed light into the convergent processes that evolved in response to thermal adaptations. [Copyright &y& Elsevier]

Abstract: With the notable exception of Antarctic icefishes, haemoglobin (Hb) is present in all vertebrates. In polar fish, Hb evolution has included adaptations with implications at the biochemical, physiological and molecular levels. Cold adaptation has been shown to be also linked to small changes in primary structure and post-translational modifications in proteins, including hydrophobic remodelling and increased flexibility. A wealth of knowledge is available on the oxygen-transport system of fish inhabiting Antarctic waters, but very little is known on the structure and function of Hb of non-Antarctic notothenioid fishes. The comparison of the biochemical and physiological adaptations between cold-adapted and non-cold-adapted species is a powerful tool to understand whether (and to what extent) extreme environments require specific adaptations or simply select for phenotypically different life styles. This study focuses on structure, function and molecular phylogeny of Hb in Antarctic and non-Antarctic notothenioid fishes. The rationale is to use the primary structure of Hb as tool of choice to gain insight into the pathways of the evolution history of α and β globins of notothenioids and also as a basis for reconstructing the phylogenetic relationships among Antarctic and non-Antarctic species. [Copyright &y& Elsevier]

Antarctic notothenloids, along with many other polar marine fishes, have evolved biological antifreeze proteins (AFPs) to survive in their icy environments. The larvae of Antarctic notothenioid fish hatch into the same frigid environment inhabited by the adults, suggesting that they must also be protected by sufficient AFPs, but this has never been verified. We have determined the contribution of AFPs to the freezing resistance of the larvae of three species: Gyinnodraco acuticeps, Pagothenia borchgrevinki and Pleuragramma antarcticum. Of the three, only P. borchgrevinki larvae are protected by high, adult levels of AFPs. Hatchling G. acuticeps and P. antarcticuin have drastically inadequate AFP concentrations to avoid freezing at the ambient seawater temperature (-.1.91°C). We raised G. acuticeps larvae and measured the AFP levels in their blood for ∼5 months post hatching. Larval serum freezing point was -1.34±0.04°C at the time of hatch; it began to decrease only after 30 days post hatch (d.p.h.), and finally reached the adult value (-2.61±0.03°C) by 147 d.p.h. Additionally, AFP concentrations in their intestinal fluids were very low at hatching, and did not increase with age throughout a sampling period of 84 d.p.h. Surviving in a freezing environment without adequate AFP protection suggests that other mechanisms of larval freezing resistance exist. Accordingly, we found that G. acuticeps hatchlings survived to -3.6±0.1°C while in contact with external ice, but only survived to -1.5±0.0°C when ice was artificially introduced into their tissues. P. antarcticum larvae were similarly resistant to organismal freezing. The gills of all three species were found to be underdeveloped at the time of hatch, minimizing the risk of ice introduction through these delicate structures. Thus, an intact integument, underdeveloped gill structures and other physical barriers to ice propagation may contribute significantly to the freezing resistance and survival of these larval fishes in the icy conditions of the Southern Ocean. [ABSTRACT FROM AUTHOR]

This study analysed the total length ( LT)-frequency distribution of mackerel icefish Champsocephalus gunnari at South Georgia and Shag Rocks from nine bottom trawl surveys at South Georgia and eight at Shag Rocks between 1987 and 2002. The estimated mean LT of age-classes 1+, 2+, 3+ and 4+ years during January were, respectively, 14·7, 23·5, 29·8 and 35·1 cm at South Georgia. Age-classes 1+, 2+ and 3+ years were 18·3, 26·2 and 33·8 cm at Shag Rocks. The derived Bertalanffy growth parameters for South Georgia were: L∞ = 51·7 cm, k = 0·27 and t0 = −0·26. The mean LT of each age-class of C. gunnari at Shag Rocks was significantly larger than at South Georgia, equivalent to c. 5 months growth, although the annual growth in LT was similar. This is further evidence that C. gunnari hatched earlier at Shag Rocks. At South Georgia, the mean LT of age-classes 1+ and 3+ years were correlated, and significantly decreased between 1987 and 2002, and were smaller following warmer summers. This decrease in the size of C. gunnari may be the result of reduced food availability linked to climate warming. [ABSTRACT FROM AUTHOR]

Nesting sites of the naked dragonfish Gymnodraco acuticeps have been identified in 15-35 m water under fast ice adjacent to McMurdo Station, making it possible to examine embryonic development and early larval growth. Egg-laying (predominantly in October) is preceded by a distinctive whirling behavioural pattern driven by the male prodding the side of the female's abdomen. The eggs (3.42 ± 0.19 mm in diameter) are laid on rocks as a single adherent layer (c. 2500 per patch). Development is unusually protracted, the first cleavage occurring after about 24 hr at about -1.9°C. Hatching occurs about 10 months post-fertilization, beginning soon after the sun rises above the horizon. During this period one of the parents may act as a guard in an attempt to keep predators at bay. Upon hatching, the larvae (12.09 ± 0.36 mm long) swim towards the surface ice where they presumably seek refuge. Yolk absorption is complete in about 15 days. Larvae (grown in aquaria at a density of 0.7 larvae l-1) display an average daily growth rate of 0.42% over nine weeks. Hatching in aquaria can occur up to 100 days in advance of that seen in the field, suggesting that under natural conditions hatching may be delayed until an appropriate stimulus (such as the return of the sun) is received. [ABSTRACT FROM AUTHOR]

Abstract: Previous research on Antarctic notothenioids has demonstrated that cells of cold-adapted Antarctic notothenioids lack a common cellular defense mechanism called the heat shock response (HSR), the induction of a family of heat shock proteins (Hsps) in response to elevated temperatures. The goal of this study was to address how widespread the loss of the HSR is within the Notothenioidei suborder and, specifically, to ask whether cold temperate non-Antarctic notothenioids possess the HSR. In general, Antarctic fish have provided an important opportunity for physiologists to examine responses to selection in the environment and to ask whether traits of the notothenioids represent cold adaptation, or whether the traits are related to history and are characteristics of the notothenioid lineage. Using in vivo metabolic labeling, results indicate that one of the two New Zealand notothenioids possess an HSR. The thornfish, Bovichtus variegatus Richardson, 1846, expressed heat shock proteins (Hsp) in response to heat stress, whereas the black cod, Notothenia angustata Hutton, 1875, did not display robust stress-inducible Hsp synthesis at the protein-level. However, further analysis using Northern blotting clearly demonstrated that mRNA for a common Hsp gene, hsp70, was present in cells of both New Zealand species following exposure to elevated temperatures. Overall, combined evidence on the HSR in notothenioid fishes from temperate New Zealand waters indicate that the loss of the HSR in Antarctic notothenioid fishes occurred after the separation of Bovichtidae from the other Antarctic notothenioid families, and that the HSR was most likely lost during evolution at cold and constant environmental temperatures. [Copyright &y& Elsevier]

Ninety-five percent of the fish species known from the Antarctic continental shelf and upper slope are acanthomorphs, i.e. spiny teleosteans. Notothenioids (suborder Notothenioidei) are acanthomorphs and so is their sister group. Unfortunately, until recently acanthomorph intra-relationships were so poorly known that it was necessary to sample all of this diversity just to search for a single sister group relationship. Using recent advances in acanthomorph molecular phylogenetics, particular properties of separate analyses and a new protocol of dataset combination, we identified a Glade that contains the sister group of notothenioids, the Percidae (perches), and a number of relatives. Among these relatives are the Serranidae (sea basses), the genera Trachinus (weeverfish), Chelidonichthys (gurnard), Scorpaena (scorpionfish), and a group composed of the Zoarcoidei (eelpouts) and the Cottoidei (sculpins) with the Gasterosteidae (sticklebacks) as their sister group. Interestingly, that Glade contains 88% of the fish species found on the Antarctic continental shelf and upper slope. The interrelationships of its components and their distribution show that the Antarctic benthic fish fauna has at least three origins. [ABSTRACT FROM AUTHOR]

The non-long terminal repeat retrotransposons Rex1 and Rex3 were identified in 13 species of Antarctic fishes from five families of the suborder Notothenioidei. Partial reverse transcriptase gene sequences were characterized for Notothenia coriiceps, Trematomus newnesi and Dissostichus mawsoni (Nototheniidae), and Gymnodraco acuticeps (Bathydraconidae). Rex1 and Rex3 both formed a notothenioid-specific monophyletic group compared to the corresponding elements from other fishes. They globally evolved under purifying selection, showing their activity during notothenioid evolution. Fluorescence in situ hybridization analysis of the chromosomal distribution of Rex1 and Rex3 was performed for several notothenioid fish species. Rex1 was generally less abundant than Rex3, which was widely scattered on the chromosomes with more intense hybridization patterns in some specific zones. Particularly, Rex3 accumulated in Chionodraco hamatus in pericentromeric areas, short arms of some pairs as well as in an intercalary band in the long arm of the Y chromosome similarly to a previously described DNA transposon. Such pattern similarities suggest the presence of autosomal and gonosomal regions of preferential accumulation for different types of repeated elements in notothenioid genomes. To the best of our knowledge, this report is the first description and analysis of retrotransposable elements in Antarctic fish genomes. [ABSTRACT FROM AUTHOR]

Hypotheses concerning the diversification of notothenioid fishes have relied extensively on estimates of divergence times using molecular clock methods. The timing of diversification of the cold adapted antifreeze glycoprotein (AFGP)-bearing Antarctic notothenioid Glade in the middle to late Miocene has been correlated with the onset of polar climatic conditions along the Antarctic Continental Shelf. Critical examination of the previous molecular clock analyses of notothenioids reveals several problems associated with heterogeneity of nucleotide substitution rates among lineages, the application of potentially inappropriate nucleotide substitution rates, and the lack of confidence intervals for divergence time estimates. In this study, the notothenioid partial gene mtDNA 125-165 rRNA (PG-rRNA) molecular clock was reanalysed using a tree-based maximum likelihood strategy that attempts to account for rate heterogeneity of nucleotide substitution rates among lineages using the penalized likelihood method, and bootstrap resampling to estimate confidence intervals of divergence time estimates. The molecular clock was calibrated using the notothenioid fossil Proeleginops grandeastmanorum. Divergence time estimates for all nodes in the PG-rRNA maximum likelihood tree were substantially older than previous estimates. In particular, the estimated age of the AFGP-bearing Antarctic notothenioid Glade predates the onset of extensive sea ice and development of polar conditions by at least 10 million years. Despite caveats involving the fossil calibration and limitations of the PG-rRNA dataset, these divergence time estimates provide initial observations for the development of a novel model of the diversification of cold adapted Antarctic notothenioid fishes. [ABSTRACT FROM AUTHOR]

The jaw musculature of notothenioid fishes is described and compared based on a total of 38 species referred to eight families of the suborder Notothenioidei. In the Notothenioidei, the adductor mandibulae, the largest and most conspicuous of the cranial muscles, is generally composed of sections A1, A2, A3 and Aw as in the generalized percoids. The morphology of the adductor mandibulae is similar in most notothenioid families except the Nototheniidae and Bathydraconidae. Notothenia, Paranotothenia and Dissostichus are clearly distinguished from the other nototheniid genera in having A3. Gymnodraco of the Bathydraconidae has a fused A1-A2 complex and the anterior element is segmented by a tendinous intersection from the A1-A2 complex. These features are unique to Gymnodraco and not seen in other bathydraconids. The Harpagiferidae and Artedidraconidae share the same apomorphy, the presence of A1β, with the Nototheniidae and have no synapomorphies with the Bathydraconidae and Channichthyidae. The character distribution, however, shows some inconsistencies with the previous classifications. Based on the limited evidence obtained in this study, the Notothenioidei can be divided into six groups and it seems reasonable to suggest a review of the current classification of the Nototheniidae. [ABSTRACT FROM AUTHOR]

The radiation of notothenioid fishes (order Perciformes) in the Southern Ocean provides a model system for investigating evolution and adaptation to a low temperature environment. The Notothenioid fishes comprising eight families, 43 genera and 122 species dominate the fish fauna in Antarctica. The diversification of the clade probably began 15–20 million years ago after the formation of the Antarctic Polar Front. The radiation was, therefore, associated with climatic cooling down to the present day temperature of −1.86 °C. Origins and Evolution of the Antarctic Biota Geological Society Special Publication No. 47, Geological Society of London. pp. 253–268). The success of the group has been closely linked with the evolution of glycopeptide and peptide antifreezes, which are amongst the most abundant proteins in blood and interstitial fluid. The radiation of the clade has been associated with disaptation (evolutionary loss of function) and recovery. For example, it is thought that the icefishes (Channichyidae) lost haemoglobin through a single mutational event leading to the deletion of the entire β-globin gene and the 5′ end of the linked α-globin gene, resulting in compensatory adaptations of the cardiovascular system. Phylogenetically based statistical methods also indicate a progressive and dramatic reduction in the number of skeletal muscle fibres (FNmax) at the end of the recruitment phase of growth in basal compared to derived families. The reduction in FNmax is associated with a compensatory increase in the maximum fibre diameter, which can reach 100 μm in slow and 600 μm in fast muscle fibres. At −1 to 0 °C, the oxygen consumption of isolated mitochondria per mg mitochondrial protein shows no evidence of up-regulation relative to mitochondria from temperate and tropical Perciform fishes. The mitochondria content of slow muscle fibres in Antarctic notothenioids is towards the upper end of the range reported for teleosts with similar lifestyles, reaching 50% in Channichthyids. High mitochondrial densities facilitate ATP production and oxygen diffusion through the membrane lipid compartment of the fibre. Modelling studies suggest that adequate oxygen flux in the large diameter muscle fibres of notothenioids is possible because of the reduced metabolic demand and enhanced solubility of oxygen associated with low temperature. At the whole animal level size-corrected resting metabolic rate fits on the same temperature relationship as for Perciformes from warmer climates. It seems likely that the additional energetic costs associated with antifreeze synthesis and high mitochondrial densities are compensated for by reductions in other energy requiring processes: a hypothesis that could be tested with detailed energy budget studies. One plausible candidate is a reduction in membrane leak pathways linked to the loss of muscle fibres, which would serve to minimise the cost of maintaining ionic gradients. [Copyright &y& Elsevier]

The involvement of nitric oxide (NO) in the branchial circulation and cardiac performance of the Antarctic hemoglobinless icefish Chionodraco hamatus was investigated using isolated and perfused head and working heart preparations. In the branchial vasculature under basal (i.e. unstimulated conditions), the nitric oxide synthase (NOS) inhibitor l-NIO (l-N5-(1-iminoethyl) ornithine, 10−5 and 10−4 M), caused a marked vasoconstriction (20%), indicating a basal nitrergic vasodilator tone, while the dose–response curve of the NO donor SIN-1 (3-morpholinosydnonimine) showed a dose-dependent vasodilator effect. Acetylcholine induced a dose-dependent branchial vasoconstriction mediated by muscarinic receptors, since the effects were abolished by pre-treatment with atropine (10−4 M). Serotonin (5-HT) induced a dose-dependent branchial methysergide-sensitive vasoconstriction which was abolished by pre-treatment with l-NIO, indicating a NO-dependent mechanism. On the isolated heart, the NOS inhibitor l-NMMA (NG-monomethyl-l-arginine) 10−4 M produced a small, but significant decrease of heart rate and, as a consequence, a decrease of power output, while the NO donor sodium nitroprusside (SNP) 10−4 M elicited increases of stroke volume, stroke work and power output, suggesting an exogenous NO-dependent positive inotropism. Exposure of the bulbus arteriosus to l-NMMA was without any appreciable effect. In contrast, SNP produced a notable relaxation of the bulbus wall with a marked increase of its stiffness, as indicated by the increase of systolic and diastolic dP/dt max (23 and 104%, respectively). [Copyright &y& Elsevier]

Although Antarctic teleosts of the suborder Notothenioidei are well studied, the status of some families remains unclear because of limited taxonomic sampling and sometimes poor statistical support from molecular phylogenies. It is true for the Bathydraconidae, the sister-family of the famous haemoglobin-less icefishes, the Channichthyidae. The present study is aimed at clarifying bathydraconid phylogeny and the interrelationships of higher notothenioid families, taking nototheniids as the outgroup. For this purpose, about 300 positions in the mitochondrial control region, 750 positions in the cytochrome b, and a matrix of morphological characters were employed for separate and simultaneous phylogenetic analyses. We conclude that (1) molecular data strongly support the split of bathydraconids into three clades, here called the Bathydraconinae (Bathydraco, Prionodraco, Racovitzia), the Gymnodraconinae (Gymnodraco, Psilodraco, Acanthodraco), and the Cygnodraconinae (Cygnodraco, Gerlachea, Parachaenichthys). Interrelationships between these three and the Channichthyidae remain unclear. Molecular data support neither paraphyly nor monophyly of the bathydraconids, while morphology leads to the monophyly of the family based on the synapomorphic loss of the spinous dorsal fin; (2) The Channichthyidae, the Harpagiferidae, and the Artedidraconidae are monophyletic families; (3) the phylogeny of the haemoglobin-less channichthyids is completely resolved and congruent with the conclusions of based on anatomical characters; (4) The present molecular results as well as other molecular studies favour the hypothesis that harpagiferids are the sister-group of artedidraconids, though our morphological matrix puts harpagiferids as the sister-group of all other families on the basis of a single character. With regard to harpagiferid relationships, it is interesting to notice that, when analysed simultaneously, morphological characters are not automatically “swamped” within molecular ones: in the tree based on the simultaneous analysis of all available data, morphological characters impose their topology on molecules. [Copyright &y& Elsevier]