[Identification and cytochemical study of an extranuclear basic protein in the spermatozoa of decapod crustaceans]. / Mise en evidence et etude cytochimique d'une proteine basique extranucleaire dans les spermatozoides des crustaces decapodes.

Extranuclear basic proteins have been detected in the capsule of the spermatozoa of three species of decapod crustaceans (Nephrops norvegicus L., Macrura; Eupagurus bernhardus L., Anomura; Carcinus maenas Penn., Brachyura). Their properties have been studied by cytochemical methods. Their position inside the capsule of the spermatozoon has been specified with the aid of the electron microscope. Present in a constant fashion in the three species cited, their relative importance is very variable. In contrast to the refringent cone of the spermatozoon of Ascaris, which contains an acid protein, ascaradine, the capsule of the spermatozoon of the three decapod crustaceans studied contains basic proteins which we propose to designate by the general term "decapodine".

Paramyosin in invertebrate muscles. I. Identification and localization.

By sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunodiffusion, we identified paramyosin in two smooth invertebrate "catch" muscles (Mytilus anterior byssus retractor and Mercenaria opaque adductor) and five invertebrate striated muscles (Limulus telson levator, Homarus claw muscle, Balanus scutal depressor, Lethocerus air tube retractor, and Aequipecten striated adductor). We show that (a) the paramyosins in all of these muscles have the same chain weights and (b) they are immunologically similar. We stained all of these muscles with specific antibody to Limulus paramyosin using the indirect fluorescent antibody technique. Paramyosin was localized to the A bands of the glycerinated striated muscles, and diffus fluorescence was seen throughout the glycerinated fibers of the smooth catch muscles. The presence of paramyosin in Homarus claw muscle, Balanus scutal depressor, and Lethocerus air tube retractor is shown here for the first time. Of the muscles in this study, Limulus telson levator is the only one for which the antiparamyosin staining pattern has been previously reported.

Paramyosin in invertebrate muscles. II. Content in relation to structure and function.

By quantitative sodium dodecyl sulfate-polyacrylamide gel electrophoresis, paramyosin:myosin heavy chain molecular ratios were calculated for three molluscan muscles:Aequipecten striated adductor, Mercenaria opaque adductor, and Mytilus anterior byssus retractor; and four arthropodan muscles:Limulus telson, Homarus slow claw. Balanus scutal depressor, and Lethocerus air tube retractor. These ratios correlate positively with both thick filament dimensions and maximum active tension development in these tissues. The role of paramyosin in these muscles is discussed with respect to the following characteristics: force development, "catch," and extreme reversible changes in length.

Determination of the presence of ceramide aminoethylphosphonate and ceramide N-methylaminoethylphosphonate in marine animals by fast atom bombardment mass spectrometry.

Phosphonosphingolipids from 15 kinds of shellfish were analyzed by fast atom bombardment mass spectrometry to determine the contents of ceramide aminoethylphosphonate (CAEPn) and ceramide N-methylaminoethylphosphonate (CMAEPn). Two pairs of ions, at m/z 126 and 140 in the positive ion mode and at m/z 124 and 138 in the negative ion mode, were used to distinguish between aminoethylphosphonic acid and N-methylaminoethylphosphonic acid in CAEPn and CMAEPn. Interestingly, mollusca in the early stage of evolution have both CAEPn and CMAEPn, while most in the middle stage have only CMAEPn and those in the highest stage have only CAEPn.

Solvent water for electrolytes in the muscle fiber of the giant barnacle.

Seven experiments are described which permit estimation of the "solvent water" or the "osmotically active water" of the dissected fiber from the giant barnacle, Balanus nubilus. Each of the first four experiments includes the measurement of a free ion activity in the myoplasm by means of a Na(+), K(+), or Cl(-) ion-specific microelectrode. The fifth experiment makes use of a membrane potential vs. [K](o) curve. The last two experiments measured fiber water and fiber volume as bath osmolarity was changed. The seven independent estimations of solvent water ranged from 0.64 to 0.72 of fiber water with a mean of 0.68. Since the extracellular space of single fibers was about 7% of fiber water, it was concluded that 25% of analyzable water was not acting as solvent for the osmotically active solutes in the myoplasm.

[Characterization of histones and chromatin of the hepatopancreas in Palaemon serratus (Crustacea Natantia)]. / Caractérisation des histones et de la chromatine de l'hépatopancréas chez Palaemon serratus (Crustacea Natantia).

The chromatin of shrimp hepatopancreas has been extracted from isolated nuclei and characterized. Nuclei were prepared in the presence of Cu++ and phenyl methyl sulfonyl fluoride in order to inhibit the nuclease and protease activities throughout the different purification steps. The purified nuclei are heterogenous in size and show a density of 1,367 g/ml determined on saccharose - glucose gradients. After washing in 0,14 M NaCl and then in 10(-2) M Tris-HCL, pH = 7,6, the nuclei were disrupted in water. The solubilized chromatin was precipitated in 0,15 M.NaCl. This chromatin is characterized by a high level of RNA (RNA/DNA = 0,38) and of non histone proteins (NHP/DNA = 0,6). The denaturation curve showed only one Tm at 69 degrees in 2.10(-4) M.EDTA. When the chromatin was extracted in the presence of staphylococcal nuclease, the Tm reached 80 degrees C. The kinetics of the digestion by the staphylococcal nuclease have been studied and show that 10 per cent of hydrolysis occurs within the first minute. The repeat length of DNA as determined with the polymers of higher order is 189 +/- 5 base pairs. The existence of nucleosomes was confirmed by electron microscopy. The superstructure of chromatin was not completely destroyed after solubilisation with a Potter. The histones were studied by gel electrophoresis after differential staining. The most important feature consists in the presence of two H1, two H2A and two H4. The acetylation levels of the histones were followed after injection of 14C-acetate in vivo. The subfraction H1, 0 was acetylated. Only one H3 was present and the two H2A fractions showed the same level of acetylation. H2B migrated faster than the H2A fractions like in Echinoderms. The two H4 fractions corresponded to two differently acetylated forms. Shrimp hepatopancreas histones were fractionated by molecular sieving on Biogel P 100 and characterized according to their electrophoretic properties as well as their amino-acid content. The amino-acid compositions of the different histone fractions were nearer to Echinoderm and Sipunculid histones, than Calf thymus homologue histones. All the fractions show a weaker basicity. The H3 fraction was the only one showing a lesser variability when compared to Calf thymus H3. The non histone proteins were extracted in 10(-2) M Tris-HCL, pH = 8 and 0.1 per cent SDS. A series of 50 proteins was detected. 80 per cent of the total amount of protein was localized in a molecular weight range comprised between 40 000 and 80 000 daltons. These proteins were compared to the histones and total proteins of sonicated chromatin solubilized by SDS in order to detect proteasic effects.

A multichannel microspectrophotometer for visual pigment investigations.

The microspectrophotometer described replaces the photomultiplier of conventional scanning systems with a multichannel detector. By eliminating scanning-related artifacts, particularly those associated with mechanical vibrations, this system makes possible ship-based microspectrophotometric studies of visual pigments of marine organisms too fragile for live transport to shore-based laboratories. The performance of the multichannel microspectrophotometer is compared with that of conventional scanning systems and absorbance spectra taken at sea on isolated rhabdoms from Euphausia pacifica are presented. Difference spectra gave a lambda max for rhodopsin of 483 nm and a lambda max for metarhodopsin of 489 nm.

Potential for employing the distribution of anomalous non-methylene-interrupted dienoic fatty acids in several marine invertebrates as part of food web studies.

A group of homologous, nonmethylene-interrupted dienoic fatty acids (NMID) recently reported in oysters has been found in several other shellfish species and also in other marine phyla. The distribution of most other "normal" fatty acids among several species of shellfish is basically similar but mantle lipids from two other molluscan species, both squid, differ radically. The squid mantle fatty acids do not include NMID, suggesting that in molluscs NMID may accumula-e primarily in filter-feeders or herbovores. The distribution of these anomalous fatty acid components in higher species suggests that they reflect invertebrates in the diet and are biochemically inert.

Localization of a marine source of odd chain-length fatty acids. I. The amphipod Pontoporeia femorata (Kröyer).

The amphipod Pontoporeia femorata (Kröyer) contains approximately equal amounts of odd chain length and even chain length fatty acids. Mature males of this species are released into the waters of Jeddore Harbour during winter months as the result of a regular reproductive cycle and become food for smelt Osmerus mordax moving into the harbor in preparation for spring spawning runs, thus accounting for the previously reported unusual fatty acid composition of these smelt. The exceptionally high levels of odd chain length fatty acids in P. femorata occur at all stages of maturity in both sexes of the animal and are found in P. femorata in other locations.

Ultrastructure and distribution of identified neurosecretory terminals in the sinus gland of the terrestrial isopod Oniscus asellus.

An ultrastructural study of the sinus gland of the terrestrial isopod, Oniscus asellus, reveals that this structure consists of two regions: the bulb, which is attached by a narrow stalk to the optic lobe, and the lateral extension, which extends from the bulb along the optic tract to the compound eye. The bulb has a distal region containing only neurosecretory terminals, and a proximal region containing terminals, glial cells, and axons that give rise to the distally located terminals. In total, the sinus gland contains five types of terminals which can be distinguished by their location and the appearance of their neurosecretory granules. Three terminal types are located in the bulb and two in the lateral extension. The size of the terminals in the bulb varies among the three types, but the number of terminals is approximately the same for each type. Conversely, the two terminal types in the lateral extension are similar in size, but differ in number. Axons of two terminal types in the bulb can be traced to the central region of the protocerebrum; axons of one terminal type in the bulb and of terminals in the lateral extension can be traced to the optic lobe.

The distribution of zinc, cadmium, lead and copper within the hepatopancreas of a woodlouse.

The distribution of metals within the hepatopancreas of Oniscus asellus (Crustacea, Isopoda) from two uncontaminated sites, and two sites contaminated with zinc, cadmium and lead, has been studied by atomic absorption spectrophotometry, light microscopy, transmission and scanning electron microscopy and X-ray microanalysis. The hepatopancreas contains two types of intracellular granule. The first type, in the S cells, are spherical granules which contain copper, sulphur and calcium. In woodlice from contaminated sites, these 'copper' granules, also contain zinc, cadmium and lead. The second type, in the B cells, are flocculent deposits which contain iron. In woodlice from contaminated sites, these 'iron' granules also contain zinc and lead. Moribund woodlice from contaminated sites have large numbers of 'copper' and 'iron' granules in the hepatopancreas and a fine deposit of zinc and lead on the membranes of the cells. There are numerous microorganisms in close association with the microvillous border of the hepatopancreas of woodlice from all four sites. Within the microorganisms of Oniscus asellus from contaminated sites, there are deposits of material which contain zinc, lead, calcium and phosphorus 'Copper' and 'iron' granules could have evolved as storage sites for essential metals to be utilized when demand from the body exceeds uptake from the food. Woodlice in contaminated sites may be able to 'detoxify' potentially harmful amounts of essential and non-essential metals by storing them in a relatively insoluble form within these granules.

The composition and nature of cuticular proteins in parasitic copepods.

1. The protein components of the cuticle of the two parasitic copepods Pennella elegans and Caligus savala were studied electrophoretically and histochemically. 2. The amino acid composition and the chemical nature of the proteins of the cuticle in the two species were also studied. 3. The fast-moving protein fractions found in the cuticle of C. savala were absent in the cuticle of P. elegans. 4. The similarities and differences in the protein components of the cuticle and their relations to the functions of the cuticle in both species are discussed.

An optimum pH for the demonstration of chitin in Periplanata americana using Lugol's iodine.

Modification of chitosan test of Campbell is suggested with a controlled procedure involving heating with saturated potassium hydroxide at 160 degrees C for a period of 45 min and subsequent washing of the deacetylated chitin in running water and application of Lugol's iodine solution (1:2:300, w/w/v) at a pH range of 1.4 to 4.0, thus replacing the use of dilute sulphuric acid. Consistent results were obtained not only with the cuticle of Periplanata americana but also with the cuticle of Emerita asiatica (crustacea).

Fate and toxicity of temephos applied to an intertidal mangrove community.

The distribution, persistence, and toxicity of the mosquito larvicide temephos was monitored following aerial applications to an intertidal mangrove community in Lee County, Florida. The amount of temephos penetrating to the mangrove floor ranged from 15 to 70% of the amount entering the upper leaf canopy, with 50-60% of that applied remaining on the mangrove leaves. Rainfall caused an additional influx of temephos from the leaves to the mangrove floor. Residues were detected in intertidal water at 2 h, but not 4 h after application. However, temephos was observed to persist in simulated tidal pools and on mangrove leaves for up to 72 h and in oysters for up to 48 h after application. Marine organisms placed in cages at 3 test sites and a control site were monitored for toxic effects. Mortality among natural mosquito larvae was simultaneously monitored. Mysids (Mysidopsis bahia) exhibited a significant mortality at one site during 1 of 3 applications monitored; however, no correlation was observed between mortality and temephos concentration in water. No significant mortality was observed for the other organisms, which included: brown shrimp (Panaeus aztecus), grass shrimp (Palaemonetes pugio), juvenile snook (Centropomis undecimalis) and sheepshead minnow (Cyprinodon variegatus).

[Sterol composition of Palaemon adspersus]. / O sterinovom sostave krevetok Palaemon adspersus.

The sterol composition of Palaemon adspersus is studied. It is established that the nonsaponifiable fraction of Palaemon adspersus contains cholesterol, 7-dehydrocholesterol, desmosterol, substance "240" and inconsiderable amounts of other still nonidentified sterols.

[Sterols of several Black Sea crustaceans]. / Issledovanie sterinov u nekotorykh rakoobraznykh Chernogo moria.

Sterol content has been investigated in lower (Artemia salina) and higher crustaceans--the isopods Idotea baltica and S phaeroma pulchellum, decapod Palaemon adspersus and several amphipods. In lower crustaceans, as well as in isopods and amphipods, no 7-dehydrocholesterol was found. This fact may be associated with peculiarities of feeding of these animals. It confirms the absence of sterol synthesis in some of the crustaceans reported in the literature. Cholesterol, demosterol and 7-dehydrocholesterol were found in shrimps.

[Preparative isolation of progesterone, testosterone and estrogens from marine invertebrate tissues]. / Preparativnoe vydelenie progesterona, testosterona i éstrogenov iz tkanei morskikh bespozvonochnykh.

Extraction and quantitative determination of progesterone, testosterone and oestrogens have been made in tissues of cephalopod molluscs and crustaceans. Extraction was made by acetone and ethyl ether. Purification was performed by thin layer chromatography on aluminium oxide and silicagel in different systems. The quantitative assay of progesterone and oestrogens was made by fluorimetric and radioimmunological techniques, that of testosterone--by spectrophotometric determination of hydrozones and radioimmunological procedure. Progesterone and testosterone were obtained from the gonads and liver of cephalopods and from the whole body of crustaceans. During sexual maturation of the animals concentration of these hormones tends to increase. Oestrone and oestradiol were found only in the gonads of cephalopods.

[Determination of fluorine in biological materials with ion-selective electrodes]. / Oznaczanie fluoru w materiale biologicznym metoda elektrody jonoselektywnej.

Fluorine was determined in faeces of experimental rats and in krill meal by the method of ion-selective electrode. Concentrated hydrochloric acid was added to the prepared samples, and then acetate buffer was added to obtain pH 5.0-5.5 The results were similar to those obtained according to AOAC. The recovery of fluorine added to the samples in amounts from 0.2 to 0.8 mg as sodium fluoride was 90 to 98%. The method may be used also for determination of fluorine level in biological materials.