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Prior understanding of Harpagifer antarcticus reproductive biology was based exclusively on macroscopic observations of its gonadal maturation cycle. Our present study with H. antarcticus specimens collected in summer of 2019 at Paradise Bay (PB), Western Antarctic Peninsula (WAP), provides a) additional reproductive parameters, and b) for first time, a histological analysis of the gonads, clarifying understanding of the spawning period of the species in the area. In females the gonado-somatic index was 5.3–19.6%, leading clutch oocytes was 0.94–1.95 mm, and total fecundity ranged between 408 and 1723 oocytes/female (X ± SD = 794 ± 328, n = 22). The histology allowed the identification of gravid females with oocytes in late vitellogenesis with evidence of ongoing hydration processes and spent females with post-ovulatory follicles. The presence of spermatozoa in mature testes indicated that the males were in developing and ripe condition. Our reproductive data for H. antarcticus at PB, validated with histology, indicate that the species was in spawning during summer, in agreement with a previous report of a late spring–summer spawning for the species at the same site. Literature data indicate that at the South Orkney Islands, H. antarcticus spawns three months earlier (May–June), probably associated with differential environmental factors between this area and the WAP. Our findings support the hypothesis that the life cycle characteristics of H. antarcticus—e.g., low fecundity and short pelagic phase—promote the isolation of populations as reflected in differences in the reproductive timing.

Antarctic coasts are highly vulnerable environments where temperature have remained very constant along millions of years. These unique environmental conditions have generated a large number of stenoic species that could be highly sensitive to future scenarios of climate change. We investigated the separate and interactive effects of increasing seawater temperature and decreasing salinity on the physiological performance of the notothenioid fish, Harpagifer antarcticus. Adult individuals were exposed to an orthogonal combination of five temperatures (2, 5, 8, 11, 14 °C) and three salinities (23, 28, 33 psu) for a 10-day period. A drastic increment in mortality was observed with seawater warming; the pattern in response to lower salinity was less clear. No fish died at the two lowest temperatures (2 and 5 °C); however, mortality increased significantly at the two highest temperatures across the salinity treatments (33.3% at 11 °C; 93.3% at 14 °C). No data were obtained at 14 °C that could be included in the physiological analyses. Ingestion and absorption rates were significantly affected by temperature and salinity, but not by the interaction of the two. Finally, we observed a negative effect of temperature but not of salinity or the interaction of both on the scope for growth of H. antarcticus. These results suggest that this species could cope with a moderate temperature increase (5 °C) in the Antarctic. However, the higher metabolic rates observed at 8 and 11 °C are associated with conditions beyond the natural thermal window of this species, representing a disadvantage in the face of climate change. Therefore, and even in the hypothetical case that H. antarcticus were able to disperse to sub-Antarctic areas such as the Magellan Region, current and projected scenarios of seawater temperatures might be unsuitable for the development of effective populations of this species. The results confirm the stenothermal nature of H. antarcticus, considering its high vulnerability to environmental changes and its limited ability to cope with the more severe global warming models projected for the Antarctic and Magellan regions for the end of the century (mainly temperature).

Over the last decades, climate change has intensified. Temperatures have increased and seawater has become “fresher” in Antarctica, affecting fish such as Harpagifer antarcticus. Thus, this study aimed to evaluate changes in the osmoregulatory response of the Antarctic notothenioid fish Harpagifer antarcticus and evaluate how it will cope with the future climate change and environmental conditions in the Antarctic, and in the hypothetical case that its geographical distribution will be extended to the Magellanes region. The present study was undertaken to determine the interaction between temperature and salinity tolerance (2 °C and 33 psu as the control group, the experimental groups were 5, 8, and 11 °C and 28 and 23 psu) and their effect on the osmoregulatory status of H. antarcticus. We evaluated changes in gill-kidney-intestine NKA activity, gene expression of NKAα, NKCC, CFTR, Aquaporins 1 and 8 in the same tissues, muscle water percentage, and plasma osmolality to evaluate osmoregulatory responses. Plasma osmolality decreased with high temperature, also the gill-kidney-intestine NKA activity, gene expression of NKA α, NKCC, CFTR, Aquaporins 1, and 8 were modified by temperature and salinity. We demonstrated that H. antarcticus can not live in the Magallanes region, due to its incapacity to put up with temperatures over 5 °C and with over 8 °C being catastrophic.

Increased levels of tissue monoaminergic neurotransmitters, as well as circulating catecholamines, appear to play a role in the regulation of the physiological responses of teleost fish. Harpagifer antarcticus is a stenothermic, Antarctic notothenioid fish. The aim of this study was to determine the effect of increased seawater temperature and decreased salinity on the levels of 5-HT, 5-HIAA, DA, and Noradrenaline in the brain, stomach, and gut of H. antarcticus. Wild-gathered fish were acclimatized to habitat conditions (2 °C, 33 PSU) prior to placement in aquaria with 4 temperatures (2, 5, 8 and 11 °C) and 3 salinities (23, 28 and 33 PSU) for 10 days. Fish exposed to 11 °C had higher levels of the brain neurotransmitters than those at 2 °C. Concomitant exposure to low salinity exacerbated the effect of exposure to 11 °C. At lower temperatures, concomitant alterations in salinity induced differential effects on brain neurotransmitters. When fish were exposed to 28 PSU, 5-HIAA, DA, and Noradrenaline levels at 5 and 8 °C presented no significant differences with those at 2 °C. In contrast, only 5HT and 5-HIAA levels in fish at 33 PSU were elevated at 5 and 8 °C respectively. Fish at 28 and 33 PSU had lower Gut 5HT levels at the 3 elevated temperatures, meanwhile fish at 23 PSU showed a biphasic effect when exposed to elevated temperatures. 5-HIAA levels decreased at 5 and 8 °C at 33 PSU. Stomach 5HT levels also showed a differential response at the 3 salinity levels when exposed to increased temperatures. At 11 °C, 5HT levels were markedly higher than those at 2 °C for fish at 33 PSU, moderately elevated for fish at 28 PSU, and lower for fish at 23 PSU, meanwhile 5-HIAA levels only increased with temperature at 33 PSU. These findings indicate that rapid exposure to alterations in temperate with or without concomitant changes in salinity is associated with differential responses in tissue monoaminergic neurotransmitter levels. The relatively high changes in neurotransmitter levels in fish exposed to moderate salinity and high temperature changes may indicate the physiological plasticity of H. antarcticus to possible changes in ocean temperature and salinity.

Physiological adaptation to increased environmental temperatures has been studied experimentally in a number of fish species, with the up-regulation of several genes identified as being associated with the process, such as the warm-acclimated protein (wap-65). This article describes the cloning and characterisation of the wap65-2 gene from the Antarctic plunderfish (Harpagifer antarcticus). The transcriptional expression of this gene in response to elevated seawater temperatures over a time course series is presented. Initially there is strong down-regulation of this gene to a maximum of 40-fold within 4 h, followed by recovery to almost control levels within 48 h, indicating that this gene does not play a role in the potential temperature adaptation of H. antarcticus.

In teleosts, the proliferation of myogenic progenitor cells is required for muscle growth and nuclear turnover. We measured the cell cycle and S-phase duration of myogenic cells in the fast myotomal muscle of two closely related Harpagifer species by cumulative S-phase labelling with 5-bromo-2'-deoxyuridine (BrdU). Harpagifer antarcticus is a stenothermal species from the Antarctic peninsula (experiencing temperatures of -2 degrees C to +1 degrees C) and Harpagifer bispinis is a eurythermal species from the Beagle Channel, Tierra del Fuego (living at +4 degrees C in winter and up to 11 degrees C in summer). Specific growth rates in the adult stages studied were not significantly different from zero. Myogenic progenitor cells were identified using an antibody against c-met. Seventy-five percent of the c-met(+ve) cells were in a proliferative state in both species. Cell cycle time was 150 h at 5 degrees C and 81.3 h at 10 degrees C in H. bispinis (Q(10)=3.4). Cell cycle duration was 35% shorter in H. antarcticus at 0 degrees C (111 h) than in H. bispinis at 5 degrees C. The predicted cell cycle time for H. bispinis at 0 degrees C (based on the Q(10) relationship) was 277 h, which was more than double that measured for the Antarctic species at this temperature. The results obtained are compatible with an evolutionary adjustment of cell cycle time for function at low temperature in the Antarctic species.

The diet of the South Georgia shag Phalacrocorax georgianus at Laurie Island, South Orkney Islands, was studied by the analysis of 420 pellets (regurgitated casts) collected from the 1995 to 1999 breeding seasons. Demersal-benthic fish were by far the main prey, followed by molluscs (mainly octopods and bivalves) and polychaetes. Harpagifer antarcticus (in 1995, 1998, 1999) and Lepidonotothen nudifrons (1996, 1997) were the most frequent prey, followed by Gobionotothen gibberifrons or Trematomus newnesi. Harpagifer antarcticus, followed by L. nudifrons or T. newnesi, was the most numerous prey in all of the seasons and predominated in mass in 1995 and 1996, followed by Notothenia coriiceps and L. nudifrons. In 1997 and 1999, G. gibberifrons and N. coriiceps were the species that most contributed to the diet whereas N. coriiceps and H. antarcticus contributed most in 1998. Our results differ from those reported for the South Orkney Islands in previous studies. These differences could be due to the use of different diet-analysis methods and to shags-related and/or fisheries-related changes in the food availability around the colonies. These alternative explanations are analysed and discussed. Fil: Casaux, Ricardo Jorge. Ministerio de Relaciones Exteriores, Comercio Interno y Culto. Dirección Nacional del Antártico. Instituto Antártico Argentino; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Ramon, Analia Ines. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Ministerio de Relaciones Exteriores, Comercio Interno y Culto. Dirección Nacional del Antártico. Instituto Antártico Argentino; Argentina

The absolute gut evacuation rate (GER) (g day−1) of Harpagifer antarcticus increased with increasing ration mass, fish mass only influenced the absolute GER at a daily ration level of 0·3% wet fish mass (approximately a maintenance ration). The relative GER (% of meal fed day−1) was also affected differently by fish and ration mass depending on the relative ration level being fed; at rations of 0·7% wet fish mass or above the relative GER decreased with increasing fish or ration mass (in such a way that the absolute GER remained constant and unaffected by fish mass). At maintenance (0·3% wet fish mass) rations the relative GER was not affected by fish size or ration mass. Thus, there appears to be a ration threshold above which the digestion physiology alters. Mass-specific GER (% g fish−1 day−1) decreased with increasing fish mass. Within a set relative ration level (% wet fish mass) an increase in fish mass decreased the mass-specific GER. At a fixed ration mass, an increase in fish mass (i.e. a reduction in the ration expressed as % fish mass) resulted in a decrease in mass-specific GER. Gut evaluation time (GET) decreased and absorption efficiency (A) increased with increasing absolute GER. The effect of ration and fish mass on the absolute and relative GER followed the same pattern irrespective of the diet, however the A and GER (% day−1 and g day−1) were higher and the GET shorter when the fish were fed shelled krill rather than amphipods.

The nitrogenous excretion rates (total ammonia nitrogen, urea, and primary amines) of plunderfish Harpagifer antarcticus were related significantly to length and to wet mass (mass exponents of 0·94, 1·01, 1·07 and 0·93 for total ammonia nitrogen, urea, primary amines, and total nitrogen, respectively). The routine total ammonia excretion rates [22·23 & 2·0 mg N kg−1 day−1 (mean±S.E.)] of plunderfish measured in Antarctica are 10–69% lower than those of comparable non-polar species. Plunderfish are ammonotelic, but the proportion of the total nitrogenous waste attributable to each category was variable between individuals. On average (ranges in parentheses), total ammonia nitrogen, urea, and primary amines accounted for c.82 (57–97), 13 (2–28), and 5 (0·6–22)%, respectively, of the total nitrogen excreted. Polar fish differ from their non-polar relatives only in the rate, and not the nature, of their nitrogenous waste excretion processes.

1. Intracellular pH (pHi) of white (fast, glycolytic) muscle in the Antarctic teleost Harpagifer antarcticus at 1°C was 7.36±0.07 (mean±s.e., n=6), as measured by 31P NMR. 2. H. antarcticus pHi and a compilation of published data indicate that the overall relationship between pHi and temperature in glycolytic muscle is biphasic, with pHi relatively independent of temperature in the range 0–17°C and inversely proportional to temperature above 17°C. 3. The data do not support the suggestion that pHi≈pN across temperatures, nor is the data consistent with alphastat regulation of pHi.

Analysis of 139 stomach contents of the Antarctic Shag (Phalacrocorax brans-fieldensis) collected at Nelson Island, Antarctica, showed that fish were the main component in the diet, followed by octopods, gammarids, euphausiids, and polychaetes. The fish No-tothenia coriiceps predominated in frequency (56%) and in mass (49%), whereas Harpagifer antarcticus was the most important by number (58%). The sizes of N. coriiceps and H. antarcticus taken by males were significantly larger than those caught by females. This dietary difference is probably due to differential prey selection related to shags' sexual dimorphism, temporal prey availability, and/or different foraging depths/areas. Observations at nests showed that females mainly foraged in the morning, whereas males foraged in the evening. No significant differences were observed between the number of daily foraging trips made by males and females, averaging 1.7 during incubation, 2.0 in early chick-rearing, and 4.6 during the late chick-rearing period. The estimated daily food intakes were 321, 315, and 758 g for females, and 421, 582, and 1,288 g for males during incubation, early and late chick-rearing, respectively.

The feeding energetics of the Antarctic spiny plunderfish (Harpagifer antarcticus) were examined with respect to the effect of both ration size and animal size. Fish of different sizes were fed single meals at one of two ration levels (2.5% wet body mass and satiation) to determine the maximum aerobic scope that could be elicited by the specific dynamic action. The excretion rates of ammonia, urea, and fluorescamine-positive substances were also monitored. Neither fish size nor ration had any effect on the factorial aerobic scope of feeding, which suggests that cellular metabolic processes associated with feeding were satiated by relatively small meals. The factorial scope in ammonia excretion was affected by both ration and fish size, indicating that respiration and excretion respond to a meal independently. The duration of the specific dynamic action response (240-390 h) increased with fish size but not ration, whereas both the time to reach the peak oxygen consumption and the duration of the ammonia response increased with ration but not fish size. The percentage of the ingested energy that was expended following feeding (the specific dynamic action coefficient) was high at low rations (approximately 56%) but lower (roughly 10%) at satiation rations. This is because the absolute energetic cost of processing a meal was largely independent of meal size. The change in O:N ratios after feeding was very ration-dependent; at low rations, O:N ratios increased, whereas at satiation rations, the O:N ratios decreased.