No abstract

The highly active squid which inhabit the pelagic zones of continental seas are characterized by high energy requirements and have been termed invertebrate athletes. In this paper, the physiological and biochemical background of muscular performance in squid from different environments is reviewed and the physiological and environmental factors limiting performance levels are addressed. Onc important factor is the highly concentrated haemocyanin which, in ommastrephid squid, and by virtue of their extreme pH dependence, helps to load oxygen efficiently in the gills and unload it fully into the tissues. Squid regulate their extracellular pH more efficiently than intracellular pH, so protecting the haemocyanin from fatal pH changes. However, a large proportion of the oxygen requirement in the mantle muscle must still be provided by oxygen uptake through the skin. Anaerobic mechanisms become involved beyond critical swimming speeds once oxygen supply to mitochondria becomes limiting. Onset of anaerobiosis also characterizes the limits of long-term tolerance to progressive hypoxia at a critical p~and to high, above-critical temperatures. In general, anaerobic energy production reflects an inability to meet oxygen demand and indicates transition to a time-limited situation. The development of energy-saving locomotion strategies therefore shifts critical thresholds and extends tolerance periods in species exposed to environmental extremes, typically in coastal areas. There, negatively buoyant squid make greater use of the fin for economical swimming, which is also advantageous because of the complexity of the environment. In Lolliguncula brevis, the rate at which anaerobic resources arc used above the critical swimming velocity is minimized by oscillating between periods of high and low pressure jets, thereby extending the period during which the animal can dive into hypoxic or warm water. However, only jet propulsion can economically attain the high velocities necessary in the open sea. Accordingly, the highest performance levels are seen in squid inhabiting the open sea, and they are only made possible by the uniform environmental parameters. Some squid may be able to operate at their functional and environmental limits, revealing a trade-off between oxygen availability, temperature, performance level and, possibly, body size.

Activities of the antioxidative enzymes superoxide dismutase (SOD), catalase, glutathione peroxidase (GPX) and glutathione reductase (GR) were measured in the cephalopods Sepia officinalis and Lolliguncula bre6is. Maximal enzyme activities were higher in gill tissue than in the mantle musculature of both species. Activities were generally lower in tissues of L. bre6is than in S. officinalis. Comparison with other ectothermic animals showed both cephalopod species to have a low enzymatic antioxidative status despite their high metabolic rate. Furthermore, changes in antioxidative enzyme activities were measured in the cuttlefish S. officinalis with increasing age. The concentrations of malondialdehyde (MDA) and lipofuscin were determined as indicators of lipid peroxidation. Investigated animals were between 1.5 months and over 12 months old. Changes of antioxidative enzyme activities with age were not uniform. SOD and GPX activities increased with age, while catalase activity declined. In contrast, GR activity remained almost unchanged in all age groups. The low level of antioxidative defense might allow for the significant age-induced rise in MDA levels in gills and mantle musculature and for the increase in lipofuscin levels in mantle and brain tissue. It might thereby contribute to increased oxidative damage and a short life span in these cephalopods.

The functional relevance of oxygen transport by hemocyanin of the Antarctic octopod Megaleledone senoi and of the eurythermal cuttlefish Sepia officinalis was analyzed by continuous and simultaneous recordings of changes in pH and hemocyanin oxygen saturation in whole blood at various temperatures. These data were compared to literature data on other temperate and cold-water cephalopods (octopods and giant squid). In S. officinalis, the oxygen affinity of hemocyanin changed at deltaP50/degrees C = 0.12 kPa (pH 7.4) with increasing temperatures; this is similar to observations in temperate octopods. In M. senoi, thermal sensitivity was much smaller (<0.01 kPa, pH 7.2). Furthermore, M. senoi hemocyanin displayed one of the highest levels of oxygen affinity (P50 < 1 kPa, pH 7.6, 0 degrees C) found so far in cephalopods and a rather low cooperativity (n50 = 1.4 at 0 degrees C). The pH sensitivity of oxygen binding (delta log P50/delta pH) increased with increasing temperature in both the cuttlefish and the Antarctic octopod. At low PO2 (1.0 kPa) and pH (7.2), the presence of a large venous oxygen reserve (43% saturation) insensitive to pH reflects reduced pH sensitivity and high oxygen affinity in M. senoi hemocyanin at 0 degrees C. In S. officinalis, this reserve was 19% at pH 7.4, 20 degrees C, and 1.7 kPa O2, a level still higher than in squid. These findings suggest that the lower metabolic rate of octopods and cuttlefish compared to squid is reflected in less pH-dependent oxygen transport. Results of the hemocyanin analysis for the Antarctic octopod were similar to those reported for Vampyroteuthis--an extremely high oxygen affinity supporting a very low metabolic rate. In contrast to findings in cold-adapted giant squid, the minimized thermal sensitivity of oxygen transport in Antarctic octopods will reduce metabolic scope and thereby contribute to their stenothermality.

No abstract

Brief squid Lolliguncula brevis are regularly exposed to fluctuating oxygen levels in their shallow coastal environment. To assess hypoxia resistance, animals were exposed for two hours to ambient oxygen partial pressures (P ) between 19.3 kPa (normoxia) and 6.6 kPa (34.2% air O 2 saturation) at 20618C. In a second set of experiments, the animals were subjected to a low P of O 2 2.860.3 kPa (14.561.6% air saturation) for 15 to 60 min. Subsequently, metabolic, energy and acid-base status were analysed in the mantle tissue. Onset of anaerobic metabolism was observed between 9.4 and 7.9 kPa (48.7 and 40.9% air saturation), reflecting the critical oxygen tension for this species. The formation of octopine and acetate indicates a simultaneous onset of anaerobic metabolism in both the cytosol and the mitochondria during progressive hypoxia. Concomitantly, an intracellular acidosis developed. During exposure to oxygen partial pressures between 19.3 and 6.6 kPa, aerobic and anaerobic processes were sufficient to maintain energy status in the mantle musculature. No significant changes in ATP and phospho-L-arginine (PLA) concentrations were observed. In contrast, both ATP and PLA levels declined significantly after 15 min at an ambient P of 2.860.3 kPa. Concomitantly, the Gibb's free energy change of ATP hydrolysis fell to a O 2 21minimum value of about 244 kJ?mol , a level suggested to reflect limiting energy availability for cellular ATPases. These results indicate that hypoxia at 2.8 kPa (14.5% air saturation) rapidly takes Lolliguncula brevis to the limits of performance. However, it is probably capable of withstanding longer periods of moderate hypoxia close to 50% air saturation (9.7 kPa), enabling the squids to cope with oxygen fluctuations in their shallow estuarine environment or to dive into hypoxic waters by use of their economic jetting strategy. Nonetheless, the critical P is O 2 considered to be high compared to other hypoxia tolerant animals, an observation likely related to the high metabolic rate of these squids.

The role of lysozyme in the immune immobilisation of Treponerna pallidurn is not yet fully understood. The T. pallidurn immobilisation assay was used to demonstrate that the immobilisation and lysis of T. palliduin in vitro by antibodies (serum, IgG fraction or IgM fraction) and complement proceed in a lysozyme-independent mode. In the presence of lysozyme the rate of immobilisation increased. In contrast with its effect on Escherichia coli, the effect of lysozyme on T. pallidurn was governed exclusively by its enzymic activity rather than by the cationic protein nature of the molecule. Lysozyme, released from stimulated phagocytes, induced formation of lysozyme antibodies in 59.6% of syphilis patients as determined by lysozyme antibody ELISA. The highest frequency was found in patients with untreated secondary syphilis, whereas untreated primary syphilis was only rarely accompanied by the presence of lysozyme antibodies. Crossreactivities between lysozyme and treponemal antigens were excluded by immunoblotting. The autoantibodies did not influence the lysozyme activity. It was concluded that the formation of lysozyme antibodies is only an epiphenomenon in the host defence against treponemal infection.

The aim of the present study was to investigate the biological role of cardiolipin antibodies during Treponema pallidum infection. Inhibition of the binding of treponemal specific antibodies at the early and late stages of infection by cardiolipin antibodies was shown in the fluorescence treponemal antibody absorption (FTA-ABS) test and T.pallidurn immobilisation (TPI) test. Incubation of treponemes with cardiolipin antibodies followed by a second incubation with treponemal specific antibodies resulted in a reduction of the titres of the FTA-ABS test and the TPI test. The findings suggest that cardiolipin antibody production should be considered as a virulence mechanism of pathogenic treponemes with the purpose of evading the host defence mechanisms.