The reproductive biology of the dab Limanda limanda (L.) in the North Sea: Seasonal changes in the ovary

Oogenesis in the dab is described in six easily identifiable stages: oogonium (Stage I), primary oocyte (II), primary vitellogenesis (III), secondary vitellogenesis (IV), tertiary vitellogenesis (V) and hyaline oocyte (VI). The annual reproductive cycle in female dab may be divided into four morphologically and histologically distinct periods: prespawning (October–January), spawning (February–April), postspawning (May–June) and resting (July–September) period.     

The karyotype of the dab (Limanda limanda L.)

In the dab ( Limanda limanda L.), the diploid number is 46 and the karyotype consists of 23 pairs of chromosomes, all of which are telocentric or subtelocentric, so that the total number of chromosome arms is 46 (AN-46).     

Seasonal changes in the common dab, Limanda limanda (L.) in Isle of Man Waters

Otolith accretion was greatest in summer when opaque bands were laid down and slowest in winter when hyaline rings were formed. Food intake was minimal in winter and maximal in summer. Liver weight was high in summer and winter and low in spring after spawning. Growth in length was higher in summer than winter. Gonad weight was greatest in winter for both sexes. Spawning occurred from February to April. After spawning males recovered earlier than females and testes ripened earlier than the ovaries. From winter to summer, the sea temperature increased from 7.5–13.0°C; the day length from 7–17h; and the salinity varied from 34.07−34.36%.     

Seasonal changes in frequency of diseases in dab,Limanda limanda,from the southern North Sea

Infestation rates of flatfishLimanda limanda from the southern North Sea (German Bight) were investigated in 1981 and 1982. At three stations, including a dumping area of acid-iron wastes northwest of Helgoland, dabs were examined for the occurrence of lymphocystis and epidermal papillomas. Marked seasonal changes of infestation rates were observed with maxima of diseased fish in March and May (epidermal papillomas 2 % in 1981, 4.1 % in 1982; lymphocystis 7.5 % in 1981, 17.2 % in 1982). Infestation minima were found from June to October. Differences in incidence rates were observed between stations in winter and spring, but not in summer. Both diseases turned out to be size-dependent. Seasonality of the two diseases is discussed in relation to spawning cycle and water quality, including pollution.     

Some observations on the seasonal distribution of dab,Limanda limanda, in the southern North Sea

The seasonal changes in the population structure of dab were observed in intervals of four to six weeks from January 1993 to November 1994 at a fixed station northwest of Helgoland. The sex distribution was characterized by a reduced number of males during autumn and winter. In the autumn of both years, an immigration of large females was evident. During long periods in spring and summer, the population structure remained relatively constant. The observations indicate that dab in the Helgoland region carry out a seasonal migration between coastal waters and the sampling area. Seasonal changes of the population structure occurred at a relatively slow rate.     

Assessment of some innate immune responses in dab (Limanda limanda L.) from the North Sea as part of an integrated biological effects monitoring

The marine flatfish dab (Limanda limanda), which lives in direct contact with contaminated sediments, is frequently used as a sentinel species in international monitoring programmes on the biological effects of contaminants. In this study, immune responses were recorded as indicators of sublethal chronic effects of contaminants, in addition to measurement of the induction of mono-oxygenase ethoxyresorufin O-deethylase (EROD) in liver cells, the inhibition of acetylcholin esterase (AChE) in muscle and a quantification of grossly visible diseases and parasites. In total, 336 dab were analysed from five sampling areas in the North Sea, including the German Bight, the Dogger Bank, the Firth of Forth, and two locations close to oil and gas platforms (Ekofisk and Danfield). When considering plasma lysozyme levels, pinocytosis and respiratory burst activity of head kidney leucocytes, a clear gradient could be observed with decreased levels in individuals collected from the Firth of Forth and locations near the oil or gas platforms compared with dab from the Dogger Bank or the German Bight. Individuals with induced EROD activity displayed reduced lysozyme and respiratory burst activities. Lysozyme levels were also reduced in dab with lymphocystis or with nematodes. The data obtained indicate that the assessment of innate immune parameters in a monitoring programme provides supplementary information about immunomodulatory effects associated with the exposure of fish to contaminants. In particular, concentrations of plasma lysozyme, which can be analysed in an easy and inexpensive assay, are considered to be an appropriate parameter for use in a battery of other bioindicators. Communicated by H. von Westernhagen and A. Diamant     

Development of 30 microsatellite markers for dab (Limanda limanda L.): a key UK marine biomonitoring species

Dab (Limanda limanda) are the principal target fish species in offshore biomonitoring programmes in the UK; however, detailed knowledge of genetic structure and connectivity among sampling locations is unavailable. Here, the isolation and characterization of 30 polymorphic microsatellite loci for dab is described. The number of alleles per locus ranged from 2 to 42, with observed heterozygosities ranging from 0.089 to 1. These loci will enable high resolution of genetic population structure and dynamics of dab around the British Isles.     

The feeding strategy of dab Limanda limanda in the southern North Sea: linking stomach contents to prey availability in the environment

The feeding strategy of dab Limanda limanda was investigated within a predefined research area in the southern North Sea for 2 years. The aims were: 1) to establish the relationship between prey availability and dab abundance, distribution, body condition and prey selection, and 2) to determine the underlying functional processes of prey choice in dab. Temporal and spatial trends of prey availability in relation to dab abundances, distribution, condition and prey selection were analysed. Temporal trends in dab condition, percentage of empty stomachs and numbers of ingested prey were apparent. In particular in the winter period 2000–2001 dab were in poor condition and prey items were only rarely found in stomachs. Similarly prey availability decreased in the environment while the abundance of dab increased. Thus, temporal changes in the nutritive state were partly thought to be attributed to density‐dependent effects. The stomach composition of dab during the two consecutive summers was mainly dominated by Pariambus typicus and Ophiura albida while in the winter period only O. albida made a significant contribution to the stomach content. Analysis of prey characteristics indicated that only prey densities in the environment significantly influenced prey choice in dab, while other characteristics such as the position of the prey in the habitat, its palatability or mobility did not have significant effects. The feeding strategy of dab thus was highly opportunistic, although trends were apparent that suggested that buried living fauna was less likely to be ingested, which was also observed in previous studies.     

Maximum daily ration and the pattern of food consumption in haddock, Melanogrammus aeglefinus (L.), and dab, Limanda limanda (L.)

Food intake was measured under conditions of continuous food availability for haddock, Melanogrammus aeglefinus (L.), and dab, Limanda limanda (L.). Haddock consumed approximately 75% of all food during the day, with a peak of feeding at dusk and an inferred peak at dawn; peaks of feeding were associated with larger meals. By contrast, dabs were mainly night-time feeders with daily consumption rates that were approximately 40% lower than those shown by haddock. Comparison of haddock maximum daily ration with published estimates of consumption in the wild indicate that wild haddock feed at between 15 and 22% of their maximum capacity. Using published data on gastric evacuation, the relationship between meal size and stomach residuum was investigated. There was no detectable relationship between these two variables.     

Gastric emptying in Limanda limanda (L.) and the return of appetite

Limanda limanda (35–225 g) fed during daytime when trained to use demand feeders under natural photoperiods. Under continuous illumination different feeding rhythms developed which were detected by periodogram analysis. Interfeeding periods (P hours) varied with body weight (W grams) and temperature (T° C) over the range 6.5–15° C as: In P=4.4 – 0.167 (In W) – 0.05 T. Daily food intake (D grams increased with both temperature and size (In D = 0.099 T + 0.579 In W – 3.49). Relative daily intake (R as % body weight, b.w) increased with temperature but decreased with size (In R = 0.099T–0.41 In W + 1.05). These changes in daily intake are primarily caused by changes in feeding frequency; the amount eaten at a meal is relatively constant save at low temperatures (6.5° C) when appetite is suppressed. Dilution of the food with kaolin (from 4.8 to 2.2 kcal/g) did not cause a compensatory increase in meal size or frequency. Gastric emptying time (GET hours) measured by X-radiography changed with body weight, temperature and relative meal size (M % b.w.) as In GET = 0.68 In M + 0.39 In W – 0.035T + 1.46. With the exception of the lowest temperature (6.5° C) interfeeding periods closely followed GET (P = 1.09 GET – 2.6) supporting the proposal that stomach fullness is a major factor controlling appetite in the dab.     

Regional patterns in prevalence of principal external diseases of dab Limanda limanda in the North Sea and adjacent areas 1992-1997

The prevalence and spatial distribution of major diseases of dab Limanda limanda in the North Sea and adjacent areas were studied in the summers 1992 to 1997. Areas covered were the North Sea, Irish Sea, northern and northeastern British Waters and the English Channel. The diseases studied were lymphocystis, epidermal hyperplasia/papilloma and skin ulceration. To standardise data, results were analysed for females >15 cm (>3 yr old). Data were subjected to median polish, and additive, extended and additive plus multiplicative models were applied to best account for effects of region and year. Annual differences in disease prevalence were low whilst differences between areas were pronounced. For lymphocystis higher prevalence was observed in the northwestern sector of the North Sea, at the northern tip of Scotland and in an area south of Iceland. Prevalence was low in the Irish Sea, the English Channel and the southern North Sea, and intermediate in the German Bight. For epidermal hyperplasia/papilloma, levels were low at Icelandic stations, in the northern Irish Sea, in the southern North Sea and the English Channel, whilst levels were high in the northwestern part of the North Sea and the German Bight. Elevated levels of skin ulceration were found on the Dogger, at 1 station in the Irish Sea (off Sellafield) and at 1 station to the south of Iceland. Lower levels were detected west of Iceland. Prevalence in all other areas was intermediate. It is concluded that a detailed analysis of available data on disease prevalence and putative causative factors is desirable and, given the good availability of data, would be a promising step forward toward elucidating possible cause and effect relationships between diseases and anthropogenic factors.     

The reproductive biology of the horse mackerel Trachurus trachurs (L.) in the North Sea and English Channel

In the horse mackerel the gametes develop asynchronously and are released in batches. The maturity cycle is characterized by a rapid post-spawning recovery period and a relatively long period of oocyte maturation. Mean gonad weight reached a peak in June and intraovarian hyaline oocytes occurred from May to August. Determination of fecundity was complicated by resorption of oocytes both before and after spawning, and by the fact that resting and developing oocytes were not clearly separated by size. An estimate of'potential fecundity'was obtained by a combination of histology and oocyte measurement. Plankton surveys showed that a major spawning area exists in the Southern Bight of the North Sea, with eggs occurring from May to late August. An estimate of annual egg production was combined with the fecundity data to give an estimate of stock size of about 500 000 tons.     

Ethoxyresorufin O-deethylase (EROD) activity in the liver of dab (Limanda limanda L.) and flounder (Platichthys flesus L.) from the German Bight. EROD expression and tissue contamination

Ethoxyresorufin O-deethylase (EROD) activity was measured in the liver of dab (Limanda limanda) and flounder (Platichthys flesus) from the German Bight (southern North Sea) and compared with muscle and liver polychlorinated biphenyl (PCB) concentrations in an attempt to relate EROD activity to PCB body burden. In none of the different datasets (species-, tissue- or matrix-dependent) was a significant (P<0.05) correlation between PCB tissue contamination and EROD activity found. Yet EROD activity was significantly correlated with polycyclic aromatic hydrocarbons (PAH) levels (phenanthrene, fluoranthene, pyrene) in muscle tissue, indicating a possible dependence of EROD expression on other ubiquitous organic contaminants, thus making it a suitable biomarker for general pollution. Received: 14 April 1999 / Received in revised form: 10 July 1999 / Accepted: 15 July 1999     

Annual changes in the nutritive state of North Sea dab

The nutritive state of dab Limanda limanda was investigated over a 2-year period at a fixed sampling site northwest of Helgoland (German Bight, North Sea), with respect to feeding habits and the accumulation of biochemical storage products. Ophiuroids formed the main weight of food organisms (50%) while polychaetes (10%), molluscs and crustaceans (<5% each) were less frequent. Feeding activity in males varied between summer and winter, while females fed more constantly. The condition factor and the hepatosomatic index showed characteristic seasonal cycles in both sexes. The glycogen content in the liver reached 40–60 mg g⁻¹ FW in summer and fell to about 10–20 mg g⁻¹ FW in late winter. Total lipids of the liver showed a distinct seasonal cycle with 400 mg g⁻¹ FW in summer and a minimum of 50–100 mg g⁻¹ FW in spring. The lipid content of the muscle ranged from 5 to 6 mg g⁻¹ FW and did not vary significantly between seasons.     

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.     

The oxygen requirements of three fish species from the German Bight: cod Gadus morhua, plaice Pleuronectes platessa, and dab Limanda limanda

The survival of cod Gadus morhua , plaice Pleuronectes platessa , and dab Limanda limanda was determined in relation to ambient oxygen saturation at 8° C and 35% m salinity. Mortalit rates were observed in fish exposed to constant oxygen levels for 24h. First mortality occurred around 60 % oxygen saturation in cod and around 30% oxygen saturation in dab and plaice. Below these thresholds mortality increased linearly with decreasing oxygen levels. If cod were infested with 1 or 2 individuals of Lernaeocera branchialis (Copepoda ), their tolerance was significantly lower; under such circumstances the incipient lethal oxygen saturation was 66 %.

Zusammenfassung

Anforderungen an den Sauerstoffgehalt im Wasser für drei verschiedene Fischarten der Deutschen Bucht: Dorsch Gadus morhua, Scholle Pleuronectes platessa und Kliesche Limanda limanda
Die Überlebensraten von Dorschen ( Gadus morthua ), Schollen ( Pleuronenes platessa ) und Klieschen ( Limanda limanda ) wurde in Abhängigkeit von dem Sauerstoffgehalt während einer 24stündigen Halterung bestimmt. Die Temperatur betrug 8° C und der Salzgehalt 35%. Eine erhöhte Sterblichkeit wurde bei Dorschen im Bereich von 60% Sauerstoffsättigung beobachtet. Für Scholle und Kliesche lag dieser Wert bei ungefähr 30% Sauerstoffsättigung. Unterhalb dieser Grenzwerte nahm die Sterblichkeit proportional zu einer Abnahme des Sauersrohgehaltes zu. Bei Dorschen, die mit Lernaeocera brancialic (Copepoda ) befallen waren, war die Toleranz gegenüber niedrigen Sauerstoffsättigungen signifikant geringer, eine erhöhte Sterblichkeit wurde schon bei 66% Sauerstoffsättigung beobachtet.     

Potential value of the comet assay and DNA adduct measurement in dab (Limanda limanda) for assessment of in situ exposure to genotoxic compounds

An in situ study of the relationship between marine contamination and genotoxic effects was performed on female dab (Limanda limanda) collected from different sites in the eastern English Channel (France) known to be contaminated by polycyclic aromatic hydrocarbons (PAHs) and polychlorobiphenyls (PCBs). DNA adducts in liver and DNA strand breaks in blood cells were determined respectively by the nuclease P1-enhanced post-labelling technique and an alkaline version of the comet assay. The extent of DNA base oxidation was also assessed for three of the six sampling sites in the study, using a comet assay in combination with a specific DNA repair enzyme, formamidopyrimidine glycosylase (Fpg).With Comet data, two groups of sites that seem in accordance with the pollution level have been distinguished. The extent of DNA strand breaks was higher in adult than juvenile female dab. From a technical point of view, comet assay sensitivity was affected by high intra-individual variability that accounted for nearly 70% of total variance (the site factor represented no more than 26%). The combined use of the comet assay and Fpg showed the presence of DNA oxidised bases in environmentally exposed dab.Although qualitative differences between the sampling sites were observed in DNA adduct profiles, no significant differences were found for total DNA adduct levels. DNA adducts did not appear to be associated with PAH exposure.Histopathological studies showed hepatic steatosis in most of the animals examined. Only one pre-cancerous lesion (an early stage of hyperplasia) was detected (associated frequency of 0.8%).