The lysozyme of the plaice Pleuronectes platessa L.

Lysozyme from the serum of the plaice, Pleuronectes platessa L., has been purified 78-fold with chitin coated cellulose.

• 1.2. Further purification on CM-cellulose yielded a single band on acrylamide electrophoresis, exhibiting lysozyme activity.
• 2.3. The quantitative amino acid composition of plaice serum lysozyme is reported.
• 3.4. The mol. wt is identical with hen egg white lysozyme.
• 4.5. A method is described for identifying fractions with lysozyme activity in polyacrylamide gels.

     

Determining abundance, distribution, and mortality of 0-group plaice (Pleuronectes platessa L.) in the Wadden Sea

Errors in estimates of year class strength and mortality among 0-group plaice can result from migrations. In order to reliably determine the number present, tidal flats and channels in a large and clearly delimited Wadden Sea area (715 km²) were sampled simultaneously using different fishing methods (by-catch, beam trawls, pushnets, and fyke nets). The numbers of 0-group plaice calculated for each depth stratum reflected changes in the distribution patterns during the complete diurnal cycle and during the course of the season and proved to be most reliable for estimating the total number in the whole area. In contrast to this, the traditional way of calculating population size by multiplying the arithmetic mean of simultaneous catches resulted in very different numbers which would entail an overestimation of mortality.     

A trypanosome from the plaice, Pleuronectes platessa L.

The results of a 12 month study of Trypanosoma platessae Lebailly (1904), a haemoflagellate parasitising the plaice, Pleuronectes platessa L., are presented. The morphological features of the trypanosome are described, with experimental investigations of the host specificity of T, platessae . The flagellate occurred at low parasitaemias in wild plaice with a seasonal variation in the numbers of infected hosts, the percentage of parasitised fish reaching a maximum of 20 % in March and declining in the summer months.
Attempts to establish T. platessae in the laboratory, by means of passage of infected blood, gave inconsistent results. Elevated levels of β-globulins were detected in the sera of T. platessae infected plaice, and it is suggested that this may be associated with antibody secretion to the parasite.     

The estimation of natural mortality in Irish Sea plaice, Pleuronectes platessa L., using tagging methods

This paper addresses the question as to whether natural mortality ( M ) can be estimated reliably in a heavily exploited fish population. Several methods, including a new opitimization technique based on Pope's cohort model, were used to estimate M from a large series of tagging experiments carried out on the Irish Sea plaice off North Wales and Ireland during the early 1960s, and off North Wales during 1979 and 1980. The assumptions underlying the new method were: (i) that the product of initial survival after tagging and the reporting rate of recaptures ( SB ) was constant for all experiments, and (ii) that the number of recoveries were log-normally distributed. Simulations showed that the estimator was robust to errors in input data. The SB values were low (0.37) but were more precise than the estimates of natural mortality for both sexes. The annual M values were 0.17 and 0.11 with standard errors 0.06 and 0.08 for males and females, respectively. The estimate of M for mature males was low, indicating a low M for older fish. The less precise estimate of M for females because of the inadequate dataset, and the higher M values obtained by applying traditional methods to the same dataset, indicated that a value of 0.2 is more appropriate for both sexes. Suggestions are made for designing and analysing tagging experiments to estimate M .     

Aspects of leucocyte migration in the plaice, Pleuronectes platessa L.

Histological studies on plaice tissues revealed mononuclear phagocytes emigrating through capillary endothelium in response to bloodborne foreign erythrocytes, with a >50% increase in circulating leucocytes within 2 h. Despite this rapid influx of leucocytes, cellular accumulation during peritoneal inflammatory responses was significantly slower than in mammals. In vitro studies with chemotaxis chambers revealed that random leucocyte migration was enhanced in the presence of endogenous chemostimulatory substances (inflammatory exudate fluid and endotoxin-activated plaice serum) although directional migration was not demonstrated. It is suggested that, compared with mammals, the slower accumulation of teleostean leucocytes at sites of injury is not due to a locomotory defect but possibly results from lower levels of endogenous chemoattractants being generated.     

Studies on the skin of plaice (Pleuronectes platessa L.)

Histological and ultrastructural examinations were carried out on the integuments of larval plaice from hatching to 60 days. The skin of the newly hatched larva consisted of a delicate epidermis overlying a fluid filled dermal space. As the fish matured this became thicker, the fibrous dermis developed and the mitochondrion rich 'chloride cells' degenerated. Around the 42nd day the eosinophil granule cell appeared in the basal layers of the epidermis and by the sixtieth day the epidermis could be considered fully developed.     

Studies on the skin of plaice (Pleuronectes platessa L.)

Eighty-nine O-group plaice from a natural population were exposed at 15°C to heavy infection by Cryptocotyle lingua cercariae. Subsequently 45 fish were retained at 15°C, whilst 44 were held at 5°C. Both groups were sampled by killing individual fish at intervals of 6,18,42 h and daily thereafter up to 710 h. Entire fish were fixed immediately in formol saline, transversely sectioned and stained by H & E, PAS, PAS-diastase, JSDB 109, Picro-Mallory, Masson's trichrome, Gram-Weigert and Alcian blue. Histopathological observations showed: (a) epidermal lesions associated with encysted metacercariae in adjacent tissues; (b) myofibrillar necrosis associated with bacteria possibly introduced by the parasite; and (c) a reactive swelling of the intermuscular septa. The progressive development of the parasite cyst and host capsule is described. Development of both was markedly inhibited at the lower temperature, but the inflammatory response at either temperature was slight. This may be evidence of a long-standing host-parasite relationship which has evolved to an advanced state of adaptation on the part of the parasite and tolerance on the part of the host.     

Studies on the skin of plaice (Pleuronectes platessa L.)

A histological, histochemical and ultrastructural examination of the skin of wild and cultured plaice was carried out, using fish from each year class from 0+ to 4+. The skin was shown to be similar in general structure to that of other teleosts but a previously undescribed cell, designated the Eosinophilic Granular Cell, a dendritic secretory cell found throughout the basal layers of the epidermis, is described. It was fixed only by formalin or dichromate, and contained numerous acidophilic granules. Melanin-bearing macrophages were observed migrating through the epithelium, but no DOPA or tyrosinase positive cells were observed by the methods used. Mast cells were very common in the dermis but were only demonstrable by special techniques. The melanophore and guano-phore systems are described and although no melanophores or melanocytes were found in the unpigmented areas of partially pigmented hatchery-reared fish, the integrity of the guanophore system was complete in such fish.     

Seasonal changes in thyroidal status in the plaice, Pleuronectes platessa L.

A chemically specific analysis for plasma thyroid hormones has been used to disclose a seasonal bimodality in the concentrations of thyroxine (T₄) and triiodothyronine (T₃) in the plaice; maxima in both hormone levels occur in winter and in summer. The ratio of T₄ concentrations to T₃ varies seasonally being at a minimum in summer. A highly significant correlation of T₄ plasma levels with the landings per unit fishing effort for the same area has been observed.     

Monoaminergic nerves in the skin of plaice Pleuronectes platessa (L.)

1. The fluorescent histochemical technique of Falck and Hillarp was applied to plaice skin. The presence of monoaminergic nerve terminals, containing predominantly stores of adrenaline, forming a plexus in and around the melanophore layer was demonstrated. 2. Such stores were enriched by noradrenaline in the presence of monoamine oxidase inhibitor, unaffected by spinal section, depleted by spinal nerve section or ligatures and abolished by reserpine. 3. The observations support the view that teleost sympathetic melanophore aggregating nerves are truly adrenergic.     

A specific ringer solution for the plaice (Pleuronectes platessa L.)

Following analysis of the serum of the blood of the plaice ( Pleuronectes platessa L.), a Ringer solution specific for this fish has been devised and tested. It has proved a satisfactory physiological saline for use in electrophysiological experiments and in the preparative procedures for electron microscopy.     

Metabolic and immunological effects of endotoxin in the plaice, Pleuronectes platessa L.

In an effort to identify factors contributing to the resistance of fish to endotoxin toxicity, the metabolic effects of an intraperitoneal injection of a Boivin preparation of E. coli lipopoly-saccharide (LPS) were studied in plaice over 4 days. Significant changes were found in serum concentrations of glucose, cortisol and non-esterified fatty acids but not in total or free cholesterol. The effect of injected LPS on the phagocytic capacity of kidney neutrophils was examined because of the possible use of LPS to promote non-specific immunity. There was no enhancement of the chemiluminescent response to the phagocytosis of unopsonized bacteria by neutrophils from plaice 24 h after LPS treatment.     

Seasonal changes in serum lipid composition of the plaice, Pleuronectes platessa L.

Seasonal changes in the major lipid classes of plaice serum were examined in fish caught off the north-east coast of Scotland. Both male and female plaice were sampled monthly between June 1983 and May 1984. Total lipid, total cholesterol, free cholesterol, triglycerides, phospholipids and non-esterified fatty acids were measured using commercial kits validated against standard reference methods. There were no significant differences in lipid levels between male and female fish and, although there were real differences between months for each lipid class, there was no regular pattern over the 12-month period. All the lipid class values, however, reached a maximum in September, which could be related to the increase in feeding after spawning. The smaller peak in February could be associated with the mobilization of lipid reserves for gonadal development and to overcome the effect of starvation during spawning.     

The effect of acute X-irradiation on the development of the plaice Pleuronectes platessa L.

Plaice eggs were irradiated with 80 kV peak X-rays at 5 % of their development from fertilization to hatching. The range of doses used was approximately 30–150 rads and the doses were measured with thermoluminescent dosimeters. The percentage hatch of eggs, survival of 3-week old larvae and survival to metamorphosis were noted. The results show that at one of the most radiosensitive stages of development, the LD₅₀/metamorphosis for plaice is approximately 90 rads. There is a steep response curve where a dose of 30 rads has little effect and where doses above 150 rads kill the majority of the larvae before metamorphosis.     

Seasonal changes in serum glucose and condition of the plaice, Pleuronectes platessa L.

Monthly sampling of plaice caught off the north-east coast of Scotland between February 1983 and May 1984 revealed a seasonal increase in condition factor, hepatosomatic index and serum glucose, which appeared to be related to the period of active feeding. The spleen indices followed a similar seasonal increase, although this was not statistically significant, while the kidney indices showed little change over the 16 months. The peak for the gonadosomatic index was in February/March 1984, but among these plaice (aged 4–5 years) there were still large numbers of immature males and females and there was no increase in 1983. Apart from the hepatosomatic index for January 1984, there was no significant difference between male and female plaice.     

The ultrastructure of plaice (Pleuronectes platessa) leucocytes

The leucocytes of plaice were examined and, at a morphological level, in addition to their response to the injection of carbon particles, they could be divided into four main types. Unlike previous reports, monocytes were seen. The neutrophil granules resembled the third granule type described in mammalian neutrophils. Monocytes and thrombocytes were the only cells to take up appreciable amounts of the carbon.     

The immunohistochemical localization of lysozyme in plaice (Pleuronectes platessa L.) tissues

The location of lysozyme in fresh plaice ( Pleuronectes platessa L.) tissues was studied using an immunofluorescent technique. An antiserum to a homogeneous sample of plaice serum lysozyme was prepared in rabbits and was shown to be monospecific by immunodiffusion and inhibition studies. This preparation, used in conjunction with fluorescent labelled sheep anti-rabbit immunoglobulin, demonstrated lysozyme in plaice neutrophils, peritoneal macrophages, gill cartilage and epithelium, spleen, kidney, skin and mucus. The distribution of lysozyme in the skin and secreted mucus was affected by the method of killing the fish.     

New microsatellites primers for plaice,Pleuronectes platessa L. (Teleostei: Pleuronectidae)

Seven microsatellite loci were developed for Pleuronectes platessa, a commercially important flatfish found throughout northern European waters. The number of alleles per locus ranged from 11 to 33 and heterozygosities from 0.74 to 0.96. Cross‐reactivity was tested against six other flatfish species. Significant amplification was found in all species from one to all seven of the loci.