Measurement of cytochrome P4501A induction in dab (Limanda limanda) and other teleosts with species-specific cDNA probes: isolation and characterisation of dab cDNA and its use in expression studies with β-naphthoflavone-treated fish

Induction of cytochrome P4501A (CYP1A) in fish is an important biomarker in marine monitoring programmes but a number of factors complicate interpretation of data based on catalytic activity. To provide additional analytical tools, we have cloned and sequenced entire (dab) and partial cDNAs (flounder, turbot, sand eel) from several fish species. A detailed analysis comparing the new sequences to those on the database (13 sequences) is presented and identifies an invariant, teleost-specific sequence (195-IVVSVANVICGMCFGRRYDH-214) which might be the basis for production of a species cross-reactive antibody. Northern and slot blots of fish RNA (sand eel, plaice, turbot, flounder and dab) showed extensive cross-species hybridisation with each of the cDNAs (sand eel, plaice, turbot, flounder and dab). The exception was turbot RNA, which only gave adequate hybridisation when the turbot probe was used. Attempts to normalise the hybridisation data to GAPDH mRNA were not satisfactory since there were significant species differences in expression of this gene and expression was suppressed (20–40%) by β-naphthoflavone treatment. The CYP1A probes indicated induction levels relative to untreated dab of: plaice (five-fold); turbot (12-fold); flounder (12-fold); and dab (10-fold). The study demonstrates the relative ease with which species-specific molecular probes can be generated and used.     

Occurrence of viral haemorrhagic septicaemia virus (VHSV) in wild marine fish species in the coastal regions of Norway

This study was aimed at determining the occurrence of viral haemorrhagic septicaemia virus (VHSV) in selected stocks of wild fish species in the coastal regions of Norway. Six cruises were undertaken covering areas which included the coastal regions of northern Norway, the Norwegian Sea, the Barents Sea, and the Skagerrak area down to the Danish border. Collected, pooled samples of internal organs (kidney, spleen and heart) from 8395 fish were examined for the presence of VHSV by inoculation on BF-2 cells. Identification of virus was performed by a standard ELISA procedure with monoclonal antibody IP5B11, which is specific for the VHSV nucleocapsid protein (N-protein). VHSV was isolated from blue whiting and Norway pout in Skagerrak. No positive samples were detected in the northern coastal regions of Norway, the Norwegian Sea, or the Barents Sea. The findings indicate a very low occurrence of VHSV in the coastal regions of Norway. Geographically, the only positive samples were obtained from fish collected in areas where VHSV has previously been found in different species of fish.     

Distribution of viral haemorrhagic septicaemia virus in wild fish species of the North Sea, north east Atlantic Ocean and Irish Sea.

A surveillance programme was initiated on the occurrence and distribution of viral haemorrhagic septicaemia virus (VHSV) in wild marine fish. Six research cruises were undertaken in an 18 mo period during 1997 and 1998, covering the North Sea, the Atlantic waters off the north and west coasts of Scotland and the Irish Sea. A total of 19,293 fish were sampled from 23 different species including cod, haddock, Norway pout, herring and sprat. Individual fish lengths were recorded and the fish were checked for lesions, haemorrhaging and other signs of disease. Pools of organ samples were taken for virus assay. The majority of fish sampled did not display clinical signs indicative of viral haemorrhagic septicaemia. A small number of cod were found with skin lesions and haddock with skin haemorrhaging. Of the 2081 organ and skin sample pools collected, 21 tested positive for VHSV by tissue culture and enzyme-linked immunosorbent assay. Seventeen of the isolates originated from Norway pout Trisopterus esmarkii, one from cod Gadus morhua (skin lesion), one from herring Clupea harengus, one from whiting Merlangius merlangus, and one from a previously unreported host species, poor cod Trisopterus minutus.     

Experimental horizontal transmission of viral hemorrhagic septicemia virus (VHSV) in Japanese flounder Paralichthys olivaceus

Infection by viral hemorrhagic septicemia virus (VHSV) has recently occurred among wild and farmed Japanese flounder Paralichthys olivaceus in Japan. In the present study, horizontal transmission of VHSV among Japanese flounder was experimentally demonstrated by immersion challenge. Exposure to a flounder isolate (Obama25) of VHSV revealed a dose-response, with higher mortality (81 and 70%) at the 2 higher exposure levels (6.0 and 4.0 log10 TCID50 ml(-1)). In a second experiment, high titers of VHSV were expressed from moribund and dead flounder based on virus detection in holding-tank waters 2 to 3 d prior to death of the fish and 1 d after death. The virus could not be detected in tank waters 2 d after death. Finally, a third cohabitation experiment in small tanks demonstrated horizontal transmission of VHSV from experimentally infected to uninfected fish.     

Species assemblages of pelagic fish embryos in the southern North Sea between 1984 and 2000

The occurrence and abundance of the pelagic eggs of southern North Sea spring-spawning fish were analysed between 1984 and 2000. Species number varied between six (1986) and 14 (1999) and was positively correlated with sea surface temperature. With one exception, dab eggs were always the most abundant and usually highly dominant. Ranking of species depended on temperature, but no significant differences in ranking between years was discernible. Although with the increase in temperature in the 1990s a change in species assemblage was evident [species belonging to the boreal-Mediterranean (Lusitanian) group became more apparent in the species assemblage] this did not lead to an increased species diversity (Shannon Index) or a change in other community parameters. It appears that the recent developments regarding spawning stock biomass of commercial North Sea fish is reflected in the declining egg abundance of the respective commercial and larger species (i.e. cod, flounder, plaice) and an increase in abundance of the eggs of small species (i.e. long rough dab, rockling) over the years. Received in revised form: 5 November 2001 Electronic Publication     

Screening for Viral Hemorrhagic Septicemia Virus in Marine Fish along the Norwegian Coastal Line

Viral hemorrhagic septicemia virus (VHSV) infects a wide range of marine fish species. To study the occurrence of VHSV in wild marine fish populations in Norwegian coastal waters and fjord systems a total of 1927 fish from 39 different species were sampled through 5 research cruises conducted in 2009 to 2011. In total, VHSV was detected by rRT-PCR in twelve samples originating from Atlantic herring (Clupea harengus), haddock (Melanogrammus aeglefinus), whiting (Merlangius merlangus) and silvery pout (Gadiculus argenteus). All fish tested positive in gills while four herring and one silvery pout also tested positive in internal organs. Successful virus isolation in cell culture was only obtained from one pooled Atlantic herring sample which shows that today''s PCR methodology have a much higher sensitivity than cell culture for detection of VHSV. Sequencing revealed that the positive samples belonged to VHSV genotype Ib and phylogenetic analysis shows that the isolate from Atlantic herring and silvery pout are closely related. All positive fish were sampled in the same area in the northern county of Finnmark. This is the first detection of VHSV in Atlantic herring this far north, and to our knowledge the first detection of VHSV in silvery pout. However, low prevalence of VHSV genotype Ib in Atlantic herring and other wild marine fish are well known in other parts of Europe. Earlier there have been a few reports of disease outbreaks in farmed rainbow trout with VHSV of genotype Ib, and our results show that there is a possibility of transfer of VHSV from wild to farmed fish along the Norwegian coast line. The impact of VHSV on wild fish is not well documented.     

Red List of macrofaunal benthic invertebrates of the Wadden Sea

Parasites were observed in medium- and small-sized fish taken from the discards of a commercial shrimper during seven different cruises in the tidal channels of the North Frisian Wadden Sea (Süderaue, North Sea) from April to September 1991. In total, 442 fish comprising four species (Sprattus sprattus, Hyperoplus lanceolatus, Ammodytes tobianus, Pomatoschistus minutus) were investigated. The parasite fauna consisted of 22 species. The parasite community structure of the 4 hosts was compared. The diet of the hosts seemed to be the main factor determining the structure of the parasite community. Other factors could not be assessed. Eight species of parasites occurred as larval stages. This indicated that fish were intermediate or paratenic hosts in their life cycle. The nematodeHysterothylacium sp. (Anisakidae) and the digeneanCryptocotyle lingua (Heterophyidea) were the dominant parasites, reaching their highest prevalence and density in sprat and sand eel. Sprat and sand eel play a very important role in parasite transmission to predacious fish and seabirds.     

Meiofauna as food source for small-sized demersal fish in the southern North Sea

Meiofauna play an essential role in the diet of small and juvenile fish. However, it is less well documented which meiofaunal prey groups in the sediment are eaten by fish. Trophic relationships between five demersal fish species (solenette, goby, scaldfish, dab <20 cm and plaice <20 cm) and meiofaunal prey were investigated by means of comparing sediment samples and fish stomach contents collected seasonally between January 2009 and January 2010 in the German Bight. In all seasons, meiofauna in the sediment was numerically dominated by nematodes, whereas harpacticoids dominated in terms of occurrence and biomass. Between autumn and spring, the harpacticoid community was characterized by Pseudobradya minor and Halectinosoma canaliculatum, and in summer by Longipedia coronata. Meiofaunal prey dominated the diets of solenette and gobies in all seasons, occurred only seasonally in the diet of scaldfish and dab, and was completely absent in the diet of plaice. For all fish species (excluding plaice) and in each season, harpacticoids were the most important meiofauna prey group in terms of occurrence, abundance and biomass. High values of Ivlev’s index of selectivity for Pseudobradya spp. in winter and Longipedia spp. in summer provided evidence that predation on harpacticoids was species-selective, even though both harpacticoids co-occurred in high densities in the sediments. Most surficial feeding strategies of the studied fish species and emergent behaviours of Pseudobradya spp. and Longipedia spp. might have caused this prey selection. With increasing fish sizes, harpacticoid prey densities decreased in the fish stomachs, indicating a diet change towards larger benthic prey during the ontogeny of all fish species investigated.     

Comparative study of growth of flatfishes on the west coast of Brittany

Lusitanian and boreal flatfish species live in the Bay of Douarnenez (Brittany, France); ten are abundant enough to be the subject of this growth investigation. In all species, females attain greater ultimate size than males, and oldest specimens are frequently female. Species are segre-gated to their longevity: those rapidly reaching their asymptotic length have the shortest life spans and also the highest values of the Von Bertalanffy parameter K .
In the bay, species of the boreal fish fauna (plaice, dab) find suitable environmental conditions for rapid growth; fish of the Lusitanian fauna (sand soles) grow slower than those living in southern areas.
In the Atlantic Ocean, the growth of the sole is better than in the North Sea and the Mediterranean.     

The host specificity of Lepeophtheirus pectoralis (Müller, 1776) (Copepoda: Caligidae)

The host specificity of Lepeophtheirus pectoralis (Müller) was examined experimentally by exposing different fish species to infection by artificially reared copepodid larvae. Copepodids which were hatched from eggs of adults parasitic on plaice ( platessae copepodids) preferred plaice to all other fishes tested, whereas copepodids which were hatched from eggs of adults parasitic on flounder ( flesi copepodids) preferred flounder to all other fish species. These behavioural differences suggest that two strains of L. pectoralis exist, one ( platessae ) adapted to plaice as its host and the other ( flesi ) to flounder. Comparison of an experimentally derived order of host preference with a table of occurrence obtained from the literature, suggests that a third strain of L. pectoralis , adapted to dab as its host, might also occur.
The process of infection by L. pectoralis copepodids is also described. It comprises a host location phase, during which the copepodid enters the habitat of its flatfish hosts and locates a host individual, and an attachment phase. The host location phase appears to be governed by changes in the activity of the copepodid and by its positively rheotactic response to water currents produced by the host. The attachment phase is probably based on the response of the copepodid to chemical factors produced by the host.     

Allozyme variation and genetic inter-relationships between seven flatfish species (Pleuronectiformes)

A total of 11 enzymes coded by 14 loci were assayed for each of 17 populations (from the north east Atlantic and Mediterranean) of seven flatfish species, including representatives of three of the four European families of the order Pleuronectiformes. Diagnostic alleles were observed for each species and there were fixed differences between the species at many loci. Thus all species were genetically distinct, although there were some common alleles. H_obs was higher than the average of a range of fish species and was also higher than that of vertebrate species as a whole. It seems that flatfish as a group may show higher levels of genetic variation than other fish. Values of genetic identity for all pairwise comparisons fell comfortably within the ranges expected. The data support the grouping of plaice and flounder into a single genus, Pleuronectes , but it is concluded that the retention of the dab in a separate genus, Limanda , is justified. A high level of genetic divergence was found between Dover and thickback soles. Genetic divergence data support the hypothesis that Pleuronectes flesus luscus in the Aegean Sea is a distinct subspecies of Pleuronectes flesus (flounder). The data show a clear separation of the Mediterranean Dover sole population from those in the Atlantic. Low genetic divergence was observed between the Aegean Sea and Atlantic brill populations. It is speculated that about 5 Mya the families Pleuronectidae and later Soleidae evolved from the ancestral Scophthalmidae.     

Effects of bottom trawling on fish foraging and feeding

The effects of bottom trawling on benthic invertebrates include reductions of biomass, diversity and body size. These changes may negatively affect prey availability for demersal fishes, potentially leading to reduced food intake, body condition and yield of fishes in chronically trawled areas. Here, the effect of trawling on the prey availability and diet of two commercially important flatfish species, plaice (Pleuronectes platessa) and dab (Limanda limanda), was investigated over a trawling intensity gradient in the Irish Sea. Previous work in this area has shown that trawling negatively affects the condition of plaice but not of dab. This study showed that reductions in local prey availability did not result in reduced feeding of fish. As trawling frequency increased, both fish and prey biomass declined, such that the ratio of fish to prey remained unchanged. Consequently, even at frequently trawled sites with low prey biomass, both plaice and dab maintained constant levels of stomach fullness and gut energy contents. However, dietary shifts in plaice towards energy-poor prey items were evident when prey species were analysed individually. This, together with a potential decrease in foraging efficiency due to low prey densities, was seen as the most plausible cause for the reduced body condition observed. Understanding the relationship between trawling, benthic impacts, fish foraging and resultant body condition is an important step in designing successful mitigation measures for future management strategies in bottom trawl fisheries.     

The round goby (Neogobius melanostomus) in the Gulf of Gdańsk – a species introduction into the Baltic Sea

In recent years, information concerning the awareness of organisms accidentally introduced into the Baltic Sea has substantially improved. Non-indigenous Estuarine and Marine Organisms (NEMO's) are hazardous for the Baltic ecosystem. Currently, about one hundred species are identified as accidentally or intentionally introduced into the Baltic Sea. Ballast waters and escape from aquaculture are the most important invasion vectors. During the last decade, an invasion of the round goby (Neogobius melanostomus) – a Ponto-Caspian fish species has been observed in the Gulf of Gdańsk. The first record of this fish in the Baltic Sea is from 1990. Early detection of the invader enabled the study of population growth and changes in the area. The first years of invasion were characterized by low numbers of individuals and a limited distribution. Later, the round goby gradually colonized all shallow waters in the western part of the Gulf of Gdańsk. Initially the fish inhabited stony and rocky habitats, but later it also occupied sandy bottoms. The round goby is now the dominating fish species in most of the shallow waters of the Gulf of Gdańsk. Two main factors account for the successful invasion of this fish in the region: the state of ecosystem at the time of the invasion and the biological features of N. melanostomus. In the late 1980s, the shallow waters of the Gulf of Gdańsk were almost devoid of piscivorous fishes. Concurrently, bivalves (a preferred prey of the round goby) have increased. Important is also parental care of laid eggs and reproductive strategy. Population growth potential enables the colonization of nearby regions. The first round gobies in the Vistula Lagoon were collected in 1999 and colonization of other Baltic Sea areas is anticipated.     

0-group flatfish growth conditions on a nursery ground (Bay of Canche, Eastern English Channel)

The settlement and growth conditions of three 0-group flatfish species, sole (Solea solea L.), plaice (Pleuronectes platessa L.) and dab (Limanda limanda L.) in the Bay of Canche, French coast of the Eastern English Channel, were investigated during summer 1997. 0-group dab were the more abundant of the three species (75.4%) followed by plaice (21%) and sole (3.6%). Analyses of fish size distribution indicated that settlement of plaice occurred before sole and dab. Growth of sole was faster than that of plaice, which in turn was faster than that of dab (0.68, 0.43 and 0.33 mm d^?1 respectively). Sole differed from plaice and dab by their larger mean length attained at the end of their first summer stay in the nursery ground. Growth performances of 0-group sole and plaice were analysed by comparing estimated growth in the field with predicted maximum growth according to temperature-growth rate models from experimental studies of growth with unlimited food supply. The data of this study suggest that plaice growth may have been limited during late summer, whereas sole growth does appear to have been determined only by water temperature.     

The feeding ecology of Pleuronectes platessa L., Limanda limanda (L.) and Scophthalmus rhombus (L.) in Carmarthen Bay, South Wales, U.K.

To assess the feeding habits of plaice, Pleuronectes platessa L., and dab, Limanda limanda (L.), within Carmarthen Bay, three sampling techniques were used: benthic trawling and surf-zone push-netting were undertaken during the feeding season for the gut contents of plaice and dab (age groups 0-IV) and of 0-group brill, Scophthalmus rhombus , to be analysed in detail, and a benthic macro-fauna grab survey covered the trawled ground to assess the distribution of food species in relation to gut contents. Plaice consumed commonly-occurring food species whereas dab took any available food. The main items were the same for both species, but secondary food preferences differed. Variations in gut contents within species, for the same sized fish, chiefly resulted from the spatial distribution of fish and not seasonal variations in the benthic macro-fauna. The length and not the age of fish primarily determined the selection of food organisms.     

Sperm adaptation in relation to salinity in three goby species

In externally fertilizing species, the gametes of both males and females are exposed to the influences of the environment into which they are released. Sperm are sensitive to abiotic factors such as salinity, but they are also affected by biotic factors such as sperm competition. In this study, the authors compared the performance of sperm of three goby species, the painted goby, Pomatoschistus pictus, the two-spotted goby, Pomatoschistus flavescens, and the sand goby, Pomatoschistus minutus. These species differ in their distributions, with painted goby having the narrowest salinity range and sand goby the widest. Moreover, data from paternity show that the two-spotted goby experiences the least sperm competition, whereas in the sand goby sperm competition is ubiquitous. The authors took sperm samples from dissected males and exposed them to high salinity water (31 PSU) representing the North Sea and low salinity water (6 PSU) representing the brackish Baltic Sea Proper. They then used computer-assisted sperm analysis to measure the proportion of motile sperm and sperm swimming speed 10 min and 20 h after sperm activation. The authors found that sperm performance depended on salinity, but there seemed to be no relationship to the species' geographical distribution in relation to salinity range. The species differed in the proportion of motile sperm, but there was no significant decrease in sperm motility during 20 h. The sand goby was the only species with motile sperm after 72 h.     

Diseases in North Sea fishes

Prior to the studies reviewed here, only lymphocystis and skeletal deformities of a variety of fish species and certain diseases of eel were known to occur in the German Bight (North Sea). From 1977 until now, 9 externally visible lesions on North Sea fishes were observed; in addition to those mentioned before, they comprise: fin rot, ulcerations, epidermal papilloma, hyperplasia, pseudobranchial tumour, eye diseases and gill swellings. With the exception of information on changes in frequencies of vertebral deformities of herring from the 1950's to the 1970's, there are no long-term data characterizing changes in frequencies of the diseases under study. For pseudobranchial tumours of cod and epidermal papilloma of dab, information is provided on occurrence and abundance. The distribution pattern of cod afflicted with pseudobranchial tumours is strongly influenced by the migratory behaviour of the fish. Epidermal papillomas of dab were more frequently found at stations within the inner German Bight than in neighbouring areas. The Bight is used for dumping of wastes from titaniumdioxide production. Further disease hot spots are areas off the Humber estuary and the British coast. Analysis of chromium in dab from the German Bight revealed elevated concentrations in epidermal tissues of specimens from the dumping area compared with that found in dab from neighbouring localities. Particulate iron was demonstrated to occur in mucous cells of dab from the dumping area. From increased levels of heavy metals with cancerogenic potential in sensitive target tissues and from increased prevalences of diseased fish in the dumping area it is concluded that these phenomena are possibly causally linked. In the vicinity of the Humber estuary high disease rates were encountered and areas with high prevalences of dab afflicted with epidermal papilloma extended over regions shown to be transport routes for persistent pollutants such as radioactive materials. It is therefore suggested that the long-range distribution of fish diseases in the southern North Sea might reflect the long-range transport of persistent pollutants.     

青岛近岸水域春、秋季鱼类群落结构

根据2004年春、秋季渔业资源底拖网调查数据,应用单元和多元统计方法分析了青岛近岸水域春、秋季鱼类群落结构。春、秋季调查分别渔获31、32种鱼类,平均每网渔获量分别为7.36kg/h和2.99kg/h,优势种类主要包括赤鼻棱鳀Thrissa chefuensis、大泷六线鱼Hexagrammo sotakii、方氏云鳚Enedrias fangi、黄鳍刺鰕虎鱼Acanthogobius flavimanus、细纹狮子鱼Liparis tanakai和长条蛇鲻Saurida elongata等。聚类分析表明,春、秋季青岛近岸水域鱼类群落都可划分为3个站位组,分别对应于北部、中部和南部调查水域。不同站位组的优势种组成变化较大,不同站位组间种类组成差异显著。分析了造成站位组内种类组成相似的特征种和造成不同站位组间相异的分歧种。不同站位组的特征种不同。赤鼻棱鳀、大泷六线鱼、黄鳍刺鰕虎鱼、方氏云鳚、星康吉鳗Conger myriaster、角木叶鲽Pleuronichthys cornutus和细纹狮子鱼是春季所有站位组间的分歧种;赤鼻棱鳀、白姑鱼Argyrosomus argentatus、大泷六线鱼、带鱼Trichiurus lepturus、短吻红舌鳎Cynoglossus joyneri、方氏云鳚、细条天竺鱼Apogonichthys lineatus、小黄鱼Pseudosciaena polyactis和矛尾鰕虎鱼Chaeturichthys stigmatias是秋季所有站位组间的分歧种。大多数特征种也是不同站位组间的分歧种,主要是由于这些种类的数量分布的空间变化造成的。青岛沿海鱼类群落结构存在明显的时空异质性。     

Drinking rates and Na+ effluxes in response to temperature change in two species of marine flatfish: dab,Limanda limanda and plaice,Pleuronectes platessa

The relationship between fish mass and drinking rate in two species of flatfish, dab and plaice, weighing between 1 and 150 g was investigated. Both plaice and dab showed increased drinking rates with increasing fish mass, although — when calculated on a weight-specific basis — the increase was negligible. Fish were acclimated to winter and summer temperatures of 9 and 14°C, respectively. In winter both species were acutely transferred to 5, 14 and 21°C and in summer to 5, 9, 21 and 25°C. Drinking rates, Na⁺ efflux and body ion content were measured. Dab showed lower drinking rates than plaice (e.g. the weight-specific drinking rates of summer-and winter-acclimated dab were 0.12±0.01 and 0.06±0.006 ml·h^-1·100 g^-1, respectively, compared to that of plaice which were 0.25±0.02 ml·h^-1·100 g^-1 in summer and 0.17±0.02 ml·h^-1·100g^-1 in winter). Summer dab exhibited decreased weight-specific drinking rates at 5, 9 and 25°C, while winter dab increased drinking at 21°C. Winter plaice also showed increased drinking at 21°C and a decrease at 5°C, but in contrast summer plaice did not increase drinking at either 21 or 25°C but showed a decrease at 5 and 9°C. Winter dab and plaice showed similar Na⁺ efflux rates but summer dab showed higher efflux at all temperatures except 5°C. The data indicates that (a) the osmoregulatory function of plaice is much weaker than that of dab at higher temperatures (>20°C) and (b) mass has a greater effect on drinking and Na⁺ efflux rates than temperature; although when calculated on a weight-specific basis neither drinking nor efflux showed any variation with fish mass suggesting that these functions occur at similar intensities across the entire weight range.Abbreviations ANOVA analysis of variance - ⁵¹Cr-EDTA ⁵¹chromium ethylenediaminetetra-acetic acid - SW sea water     

Oxygenation properties of haemoglobins from the flatfish plaice (Pleuronectes platessa) and flounder (Platichthys flesus)

Summary The oxygen binding properties and some haematological data of haemoglobins of plaice (Pleuronectes platessa) and flounder (Platichthys flesus) were measured and compared, with the view of discerning their adaptations to the benthic habitat.Compared to plaice, flounders posses higher haematocrit and haemoglobin levels, and their haemoglobin has a higher oxygen affinity, smaller Bohr and Root effects and appears to have a greater ATP sensitivity. At pH 7.6, the half-saturation oxygen tensions,P ₅₀, of stripped plaice and flounder haemoglobins amount to about 5.9 and 4.2 torr, and the Bohr shifts (logP ₅₀/ pH) to –0.51 and –0.34, respectively. The species differences in the oxygen binding patterns occur after stripping the haemoglobin in solution, from dissolved ions and thus appear to be inherent in the pigment molecules. The differences suggest that the haemoglobin of flounder is better adapted to hypoxic and hypercarbic environments than plaice haemoglobin, in accordance with the more inshore occurrence of the former species.The molar ratios of ATP to haemoglobin tetramers are low in plaice and flounder (about 1.6); in plaice this ratio corresponds to that where the cofactor effect is most pronounced. In contrast to previous evidence for an adaptive reduction in the temperature dependence of the haemoglobin of flounder but not of plaice from the Baltic Sea (Friedrich, 1935), the same temperature effects were found in representatives of the two species both from the Dutch North Sea and the Danish Kattegat.