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.     

Temperature and somitogenesis in embryos of the plaice (Pleuronectes platessa)

The rate of somitogenesis was observed in plaice embryos reared at 5 and 12°C. Somite formation occurred relatively later in development at 12 than at 5°C. At any given stage of development 5°C plaice embryos had significantly more somites than embryos reared at 12°C.     

Stress and water balance in the plaice Pleuronectes platessa

Summary Drinking and branchial water permeability have been measured in plaice, and the effect of handling stress on these and on water balance is reported. Stress markedly disturbs water balance, with a weight loss of 1.6% over 7 h, and a recovery period exceeding 19 h. Depression of the normal rate of ingestion accounts for about 14% of this imbalance. Stress-induced urination is not a significant factor. Branchial diffusional water permeability is more than trebled in response to handling, but a measure of accommodation is seen with repeated handling. The ratio of osmotic to diffusional branchial water permeability is about 2.5 and is not appreciably affected by stress. Oxgen uptake is enhanced in response to stress, and this is discussed in relation to the effects of stress on water balance.     

Sexual maturation in triploid male plaice (Pleuronectes platessa) and plaice x flounder (Platichthys flesus) hybrids

Triploid male plaice, Pleuronectes platessa , and plaice x flounder, Platichthys flesus , hybrids produced from cold-shocked eggs were reared to maturity at three years of age. The testes of triploid hybrids were similar in macroscopic appearance and size to those from control fish but in histological section spermatocyte and spermatid cell formation was grossly abnormal. In two fish the majority of cells appeared to be aborted spermatids and spermatocytes although there were some spermatozoa with enlarged heads and thick bent flagellae.
The testes of mature triploid plaice were similar to controls both in shape and size and in histological section. Spermatocyte and spermatid cell formation was normal, apart from an increase in cell and nuclear size, and fully motile spermatozoa were produced from several fish. Plaice eggs fertilized with semen from triploids showed a slight reduction in fertility compared with controls but developed normally to gastrulation with little mortality. Beyond gastrulation there was increasing mortality associated with abnormalities in the appearance of embryos. Few hatched and none of these survived longer than 48 h. The value of triploidy in fish farming is briefly discussed.     

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.     

Leucocytes and related cells in the plaice Pleuronectes platessa

The leucocytes and related cells of the blood of plaice were examined morphologically and their various functions assessed using a number of procedures to identify phagocytosis histochemical reactions and antibody responses. Four morphologically different types of thrombocytes were identified in addition to lymphocytes, plasma cells, monocytes, macrophages and one type of granulocyte which histochemically resembled the mammalian neutrophil. The evolution and development of the cells was also investigated and the various stages described.     

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.

     

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.     

Genetic analysis of enzyme polymorphisms in plaice (Pleuronectes platessa).

Genetic analysis was performed on five enzyme systems (G3PDH; GPI-A; GPI-B; PGM; MDH-A) in plaice (Pleuronectes platessa) collected in spawning condition from the North Sea. Conventional crosses, induced gynogenesis and induced triploidy were performed. The data conclusively demonstrated the inheritance of isozymes by co-dominant alleles at individual loci for each system. No linkage was observed but tests did not include MDH nor the possibility of linkage between G3PDH and GPI-A. Some anomalous segregation ratios were observed, particularly a deficiency of heterozygotes for GPI-A, but the data were largely in conformity with Mendelian expectations. At the PGM locus, five independent anomalous individuals were scored and interpreted as mutations with a mutation rate of 1.1 X 10(-3) per gamete. Recombination with the centromere was assessed and induced triploidy and cross-over values of 41 per cent for PGM, 19 per cent for MDH-A and 9 per cent for GPI-B were derived on the assumption of complete interference. Amongst the parent fish, genotypic and phenotypic frequencies were largely consistent with the expectations of the Hardy-Weinberg Law, and allelic frequencies were not significantly different between year of collection or location of collection ground.     

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.     

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.     

Otolith oxygen and carbon stable isotopes in wild and laboratory-reared plaice (Pleuronectes platessa)

The relationship between water temperature, growth rate, and otolith isotopic ratios was measured for juvenile plaice (Pleuronectes platessa) reared at two temperatures (11 and 17°C) and two feeding regimes (1 and 3 prey items·ml^?1). The otolith isotope ratios in individual fish ranged from ?2 to ?4 for carbon isotope ratios (δ¹³C) and from 0.2 to 1.9 for oxygen isotope ratios (δ¹⁸O). The otolith oxygen isotope ratios were significantly affected by water temperature, but not by feeding level, and there were no significant synergistic effects. The fractionation of oxygen isotopes during otolith growth was independent of individual growth rate. Carbon isotope ratios were not significantly affected by food ration or water temperature, but were related to fish growth rate. The carbon isotope ratios were negatively correlated with fish length in the colder water treatments, and tended to increase with fish length in the warm water treatments. The laboratory-determined relationship between otolith oxygen isotope ratio and water temperature was applied to individuals of five species (plaice, cod, whiting, haddock, gurnard) collected in a single trawl sample. The otolith derived temperatures often overestimated measured water temperatures. The difference between real and estimated water temperatures varied between species, and the closest fit was for field-caught plaice.     

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.     

Isolation and properties of sarcoplasmic reticulum from the fast muscles of plaice Pleuronectes platessa L.

This paper describes a method for the isolation of highly purified sarcoplasmic reticulum from plaice fast muscle. The interrelationships of pH, KCL, Ca²⁺, Mg²⁺, ADP and temperature have been investigated. Protein composition of plaice white muscle sarcoplasmic reticulum was found to be comparable to that described for rabbit fast muscle, with a major component of 100 000 daltons. Arrhenius plots of the Ca²⁺-AT Pase are linear over the range 0–30°C. Activation enthalpy (60±1.5 kJ/mol) was found to be independent of KCl concentration. The calcium concentration required to give half maximal activation of the AT Pase (K_Ca) was found to decrease with increasing temperature, from a maximum of 1.7 μm at 0°C to 0.55 at 20°C.     

Isolation of substances with vascular permeability-increasing activity from the skin of the plaice (Pleuronectes platessa L.)

A methanolic extract of plaice skin, from which lipids had been removed, was chromatographed on alumina, eluted with decreasing concentrations of ethanol. Only the 60% ethanol fraction exhibited smooth muscle activity, with bradykinin-like properties. The 20% ethanol fraction increased vascular permeability in rat skin, as measured by dye-leakage. This was not due to the degranulation of mast cells. Intradermal injection of either fraction into the plaice caused localized erythema.