Induced inflammatory process in the Antarctic fish Notothenia neglecta

We studied the ability of the Antarctic fish Notothenia neglecta to conduct an induced inflammatory process at 0°C. Indian ink was injected and a cotton suture thread was implanted into muscle of different groups of fish. After 1–2 days of Indian ink injection, the ink was diffused in the perimysium and there was hemorrhage and cellular infiltrate composed mainly of macrophages A (with few and small lysosomes) and neutrophils; after 7–15 days, there were macrophages A and some macrophages B (cytoplasm clear, lamellar cytoplasmic system forming interdigitations); after 30 days, there was Indian ink in the interior of macrophages A. The suture thread process takes place in two phases: the first (up to 7 days) with predominance of macrophages A and few neutrophils, and the second (15–30 days) with predominance of macrophages B. It can be concluded that N. neglecta is responsive to irritant stimulus with inflammatory process indicating adaptation to the antarctic environment. Received: 22 July 1997 / Accepted: 27 April 1998     

Bacterial flora of newly caught Antarctic fish Notothenia neglecta

Summary The heterotrophic bacterial flora of Antarctic fish Notothenia neglecta was studied in Potter Cove (King George Island, South Shetland Islands). Quantitative and qualitative analysis of aerobic bacteria from sea water, skin, stomach and intestine were carried out. In addition, anaerobic flora of stomach and intestine was studied and compared. Pseudomonas was dominant in sea water samples but, neither skin nor digestive tract samples showed to be rich in this genus. Stomach flora showed variable results between samples. The gut flora was composed almost exclusively of Vibrio. Despite extreme environmental conditions this Antarctic fish show an intestinal indigenous microflora very similar to warmer waters fish. Several factors, that we discussed, could be determining the Vibrio dominance in the gut of marine teleosts.     

Comparison of fatty acid composition in nine organs of the sympatric Antarctic teleost fish species Notothenia coriiceps and Notothenia rossii (Perciformes: Nototheniidae)

Fatty acid (FA) composition of nine organs from two closely related Antarctic fish species, Notothenia coriiceps and Notothenia rossii, was determined through gas chromatography with flame ionization detection. A data set for each species was obtained using major FA profiles from specimens caught in the sea waters of Admiralty Bay during the summer season. The FA profiles for both species are overall similar, but organ peculiarities have been found, which could reflect metabolic specificities and feeding habits between species. With the exception of liver, the most abundant FA in organs was the n?3 polyunsaturated FA. The total n?6 polyunsaturated FAs were minor components in all evaluated organs. Palmitic acid was identified as the major saturated FA, whereas oleic acid was the most represented of the monounsaturated FA in almost all assessed organs of both species. The n? 3/n? 6 ratios of all organs were higher than 3.5. Differences in individual FA and FA metabolic profiles of some organs observed between N. coriiceps and N. rossii suggest specific requirements in the mobilization, transport, incorporation, and/or catabolism of lipids that were reinforced by differences on some FA ratios expressing the activity coefficient of enzymes implicated on the FA pathway flux.     

Gill morphometrics of two Antarctic fish species Pleuragramma antarcticum and Notothenia gibberifrons

Summary Gill dimensions of 27 juvenile and adult Pleuragramma antarcticum from the southern Weddell Sea and of 28 juvenile and adult Notothenia gibberifrons from the South Orkney and South Shetland Islands were estimated. The unit gill area (UGA) of P. antarcticum ranged from 75 to 167 mm²/g (mean=105); gill area index (GAI) and water-blood distance (WBD) were found to be 1.38 cm² and 3.3 m, respectively. The exponent d_g in the relationship total gill area to weight was found to be 0.90. It is concluded that P. antarcticum belongs to sluggish species and, although pelagic, its routine energy costs tend to be low. However, the closely packed lamellae (NL/mm=21) indicate more active behaviour in comparison with N. gibberifrons. A preliminary estimation of growth parameters (P, k) is presented. The sluggish behaviour of N. gibberifrons, as expected from its benthic mode of life, is reflected by the gill parameters: the very low UGA ranged from 39 to 118 mm²/g; GAI and d_g were found to be 0.73 cm² and 0.98, respectively. Comparing gill dimensions and general respiration characteristics of fish from antarctic, temperate and tropical waters it is concluded that antarctic fish have increased their scope for activity.     

Karyotypes of two Antarctic fishes,Notothenia gibberifrons andNotothenia coriiceps neglecta

The chromosomes of two species of Antarctic fishes,Notothenia (Gobionotothen) gibberifrons andNotothenia (Notothenia) coriiceps neglecta, were prepared by the air-drying method at the Polish Antarctic Station “Henryk Arctowski” during the austral summer 1984–1985. ForN. (G.) gibberifrons the diploid number is2n = 46 consisting of 2 metacentric (m) pairs, 1 submetacentric (sm) pair and 20 telocentric (t) or subtelocentric (st) pairs. ForN. (N.) coriiceps neglecta the diploid number is 2n = 22 consisting of 9 m pairs, 1 sm pair and 1st pair. Some aspects of karyological evolution of these fishes are discussed.     

Stability and conformational dynamics of metallothioneins from the antarctic fish Notothenia coriiceps and mouse

The structural properties and the conformational dynamics of antarctic fish Notothenia coriiceps and mouse metallothioneins were studied by Fourier-transform infrared and fluorescence spectroscopy. Infrared data revealed that the secondary structure of the two metallothioneins is similar to that of other metallothioneins, most of which lack periodical secondary structure elements such as alpha-helices and beta-sheets. However, the infrared spectra of the N. coriiceps metallothionein indicated the presence of a band, which for its typical position in the spectrum and for its sensitivity to temperature was assigned to alpha-helices whose content resulted in 5% of the total secondary structure of the protein. The short alpha-helix found in N. coriiceps metallothionein showed an onset of denaturation at 30 degrees C and a T(m) at 48 degrees C. The data suggest that in N. coriiceps metallothionein a particular cysteine is involved in the alpha-helix and in the metal-thiolate complex. Moreover, infrared spectra revealed that both proteins investigated possess a structure largely accessible to the solvent. The time-resolved fluorescence data show that N. coriiceps metallothionein possesses a more flexible structure than mouse metallothionein. The spectroscopic data are discussed in terms of the biological function of the metallothioneins.     

Hemoglobin from the Antarctic fish Notothenia coriiceps neglecta. 1. Purification and characterisation

Antarctic fishes live at a constant temperature of -1.8 degrees C, in an oxygen-rich environment. In comparison with fishes that live in temperate or tropical waters, their blood contains less erythrocytes and hemoglobin. A study was initiated on the structure and function of Antarctic fish hemoglobin. The erythrocytes of the Antarctic benthic teleost Notothenia coriiceps neglecta, of the family Nototheniidae, have been shown to contain two hemoglobins, accounting for about 90% and 5% of the total content. These hemoglobins have been isolated, and obtained in crystalline form. They are tetramers and contain two pairs of globin chains. The globin chains of each hemoglobin have been purified and characterised. The two hemoglobins appear to have one of the two globin chains in common. The Root and Bohr effects have been investigated in erythrocytes, 'stripped' hemolysates and pure hemoglobins, indicating that the functional properties are finely regulated by pH and allosteric effectors.     

The population dynamics of Notothenia rossii from South Georgia (Antarctica)

Summary The population dynamics of nearshore juvenile Notothenia rossii were studied at South Georgia during 1978 and 1979. Mortality of the population was calculated from length-class abundance of fish sampled by trammel nets, and the population size estimated using a mark and recapture technique. Instantaneous natural mortality in the nearshore population was high (M=0.78) although other author's estimate for the mortality in the offshore population was only 0.35. Therefore it is likely that no single estimate of mortality will apply to fish of all ages. The population density of juvenile N. rossii was about 11 g m^-2 nearshore. The size of the offshore population was estimated by calculating survival from the nearshore age classes. The data suggest that the population around South Georgia has been reduced by overfishing to about one tenth of its previous unexploited level. Yield could have been increased by fishing less intensively, and the population maintained at or near its initial level if recruitment remained constant.     

Fibre types in the locomotory muscles of an antarctic teleost,Notothenia rossii

Summary The metabolic and structural differentiation of locomotory muscles of Notothenia rossii has been investigated. In this species sustained locomotion is achieved by sculling with enlarged pectoral fins (labriform locomotion), whilst the segmental myotomal muscle is reserved for burst activity. Red, white and subepidermal fibres can be distinguished in the trunk by histochemical and ultrastructural criteria. The main pectoral muscle (m. adductor profundus) consists entirely of red fibres. These three main fibres types show differences in histochemical staining profiles, capillarization, myofibril shape and packing, and lipid and mitochondrial content. The fractional volume of mitochondria amounts to 38% for pectoral, 30% for red myotomal and 1.9% for white myotomal fibres. Enzyme activities of red pectoral muscle are consistent with a higher potential for aerobic glucose and fatty acid oxidation than for the red myotomal fibres. Mg²⁺ Ca²⁺ -myofibrillar ATPase activities are similar for red pectoral and myotomal muscles and approximately half of those white fibres. Specialisations of N. rossii muscles associated with labriform swimming and locomotion at Antarctic temperatures are discussed.     

Age determination of the Antarctic fishes Champsocephalus gunnari and Notothenia rossii marmorata from South Georgia

Summary Investigations were performed on otoliths from Champsocephalus gunnari and Notothenia rossii marmorata as part of an effort to study the population dynamics of Antarctic fishes. Examination of otoliths by scanning electron microscope (SEM) revealed internal microincrements which were similar to daily increments described for other fish species. No easily identifiable annuli could be discerned and fishes were aged by counting microincrements. Multiple regression analyses relating age to otolith morphometrics and fish size, provided a simpler ageing method. It was determined with these techniques that C. gunnari grew to 50 cm in 15 yrs and N. rossii marmorata grew to 50 cm in 6.5 yrs. The lack of larger-sized individuals for N. rossii marmorata may be indicative of recent overfishing with only the younger age groups remaining. It appears that length-at-age data determined on a yearly basis, could provide valuable scientific and management information for these species.     

Timescale of ovarian maturation in Notothenia coriiceps; evidence for a prolonged adolescent phase

Ovarian maturation in Notothenia coriiceps is known to include a period during which yolk deposition begins to take place. This 'adolescent' phase was thought originally to last for about 1 year. The results from a 2–year study at Signy Island indicate that this adolescent phase lasts for about 4 years. There is also some evidence to indicate that not all sexually mature fish spawn each season. Two reproductive strategies are considered, one where adverse conditions are accompanied by a failure to spawn and an alternative where fecundity is controlled by the number of small oocytes which ripen.     

Specific immunity proteomic profile of the skin mucus of Antarctic fish Chionodraco hamatus and Notothenia coriiceps

The white-blooded Antarctic icefish is the only known vertebrate lacking oxygen-transporting haemoglobins. Fish skin mucus, as the first line of defence against pathogens, can reflect fish welfare. In this study, we analysed the skin mucus proteome profiles of the two Antarctic fish species, the white-blooded Antarctic icefish, Chionodraco hamatus, and the red-blooded Antarctic fish, Notothenia coriiceps, unfolding the different proteins by liquid chromatography coupled with tandem mass spectrometry isobaric tags for relative and absolute quantitation (iTRAQ) technology. Of the 4444 totally identified proteins, 227 differentially expressed proteins (DEPs) were found in the comparison between C. hamatus and N. coriiceps, of which 121 were upregulated and 106 were downregulated in the icefish. In the Kyoto Encyclopedia of Genes and Genomes pathway annotation, we found two pathways “Legionellosis” and “Complement and coagulation cascades” were significantly enriched, among of which innate immune candidate proteins such as C3, CASP1, ASC, F3 and C9 were significantly upregulated, suggesting their important roles in C. hamatus immune system. Additionally, the DEP protein–protein interaction network analysis and “Response to stress” GO category provided candidate biomarkers for deep understanding of the distinct immune response of the two Antarctic fish underlying the cold adaptation.     

Stress response in the Antarctic teleosts (Notothenia neglecta Nybelin and N. rossii Richardson)

Blood chemistry and haematological parameters have been determined in the Antarctic teleosts, Notothenia neglecta Nybelin and Notothenia rossii Richardson at 2° C. Samples were taken using chronically implanted dorsal aortic cannulae following a minimum of 24–36 h recovery. Broadly similar results were obtained for the two species. In N. neglecta , routinely active specimens had high values of arterial pH (7.81) and PO ₂ (9.26 kPa), and modest haemoglobin levels (5.6 g dl⁻¹) relative to temperate species. Following 3 min strenuous activity there was a decrease in arterial pH (7.63) and a small rise in lactate from 0.41 to 0.68 mm, but no significant change in the calculated net metabolic acid load (δH_m+). PaO₂ and PaCO₂ varied inversely during exercise, and oxygen content declined by 22%. pHa and most other haematological parameters returned to routine values between 1–3 h post-exercise. The results suggest that the major effect of strenuous activity in Notothenia spp. is a respiratory, rather than a metabolic acidosis.     

Metal (Fe, Zn, Mn and Cu) levels in the Antarctic fish Notothenia coriiceps

During the 1994/1995 Antarctic summer 20 Antarctic cod (Notothenia coriiceps) of both sexes were collected at King George Island, South Shetland Islands, Antarctica, to determine the levels of Fe, Zn, Mn and Cu in muscle, liver, gonad and kidney. Metal levels were assessed using the inductively coupled plasma-atomic emission spectrometry (ICP-AES) technique. The levels of Fe, Zn, Mn and Cu observed in both sexes of N. coriiceps were significantly lower in muscle tissue than in liver, kidney and gonad; the livers of males and gonads of females had the highest levels. The differences between the sexes were significant for Zn and Mn in gonads and for Cu in liver. Despite the modest size-range of the specimens, the Cu levels in liver, muscle and kidney were significantly inversely correlated with female mass (P < 0.05) as was gonad Cu level and male body length. Fe mainly presented the highest concentrations and Cu and Mn the lowest ones. Mean muscle metal levels are similar to those of hake, cod, herring and mackerel; however, the level of Fe was slightly higher. Accumulation may occur for Fe, Zn and Mn in liver, kidney and gonad, which have higher levels than those of most prey. Received: 21 October 1997 / Accepted: 1 February 1998     

Ultrastructural and biochemical changes in the muscle tissue of Notothenia nudifrons L. during prolonged cold preservation

During initial stages freezing and conservation of Notothenia nudifrons L. muscle tissue are accompanied with nonspecific changes in the intracellular membranous system of fibers, in their lipid and protein composition. Certain connections between development of destructive changes in mitochondrial membranes, sarcoplasmic reticulum, T-system and intensity of accumulation of free fatty acids and products of peroxide oxydation of tissue lipids are revealed. Manifestation of hydrolitic enzymes activity coinsides, in time, with formation of autophage vacuoles. Transformation of the latter into residual bodies is in accord with a decreased enzymatic activity. A supposition is made concerning the mechanism of the muscle tissue destruction in the process of a prolonged conservation in cold.     

Solution structure of MT_nc,a novel metallothionein from the Antarctic fish Notothenia coriiceps

The structure of [113Cd(7)]-metallothionein (MT_nc) of the Antarctic fish Notothenia coriiceps, the first three-dimensional structure of a fish metallothionein, was determined by homonuclear 1H NMR experiments and heteronuclear [1H, 113Cd]-correlation spectroscopy. MT_nc is composed of an N-terminal beta domain with 9 cysteines and 3 metal ions and a carboxy-terminal alpha-domain with 11 cysteines and 4 metal ions. The position of the ninth Cys of the alpha domain of MT_nc is different from the corresponding Cys of mammalian MTs. As a result, the last CXCC motif in the mammalian MT sequence becomes CXXXCC in the fish MT. This difference leads to a structural change of the alpha domain and, in turn, to a different charge distribution with respect to that observed in mammalian metallothioneins.