Reproductive strategy of leafscale gulper shark Centrophorus squamosus and the Portuguese dogfish Centroscymnus coelolepis on the Portuguese continental slope

Recent data are presented on leafscale gulper shark Centrophorus squamosus and Portuguese dogfish Centroscymnus coelolepis collected during an extended sampling programme carried out at mainland Portuguese landing ports. Although there are some features common to all deep-water squaliform sharks, the two species have different reproductive strategies: C. squamosus has a lower fecundity and larger total length ( L _T) at first maturity than C. coelolepis . Despite the scarcity of pregnant C. squamosus in the samples, the L _T at birth was estimated at c. 440 mm. For C. coelolepis the results suggest the existence of a non-annual reproductive cycle with two breeding seasons in each year.     

Chemical characterisation of eggs from deep-sea sharks

The chemical characterisation and nutritional value of eggs from the five deep-sea sharks leafscale gulper shark (Centrophorus squamosus), greater lantern shark (Etmopterus princeps), longnose velvet dogfish (Centroscymnus crepidater), Portuguese dogfish (Centroscymnus coelolepis) and black dogfish (Centrocyllium fabricii) captured at Hatton Bank in the North Atlantic were examined. The chemical composition was quite similar for all the eggs studied. The dominant fatty acid in all the eggs was the monounsaturated fatty acid C18:1, which varied from 27-39%. The eggs had a relatively high content of C16:0 (13.0-18.5%) and C22:6n-3 (10.3-15.1%). The two main lipid classes in the eggs were triacylglycerols (36-55%) and phospholipids (34-41%). The eggs had high amounts of vitamin A and E. The shark eggs were particularly rich in the amino acids aspartic acid, glutamic acid, leucine and arginine.     

Scavenging interactions between the arrow tooth eel Synaphobranchus kaupii and the Portuguese dogfish Centroscymnus coelolepis

A scavenging interaction between the arrow tooth eel Synaphobranchus kaupii and the Portuguese dogfish Centroscymnus coelolepis, both ubiquitous components of fish assemblages at bathyal depths, was observed. Using a baited camera between 1297 and 2453 m in the eastern Atlantic Ocean continental slope, it was shown that despite consistently rapid arrival times of S. kaupii (<5 min), their feeding bouts (indicated by acute peak in numbers) did not take place until shortly after C. coelolepis arrived and removed the exterior surface of the bait (skipjack tuna Katsuwonus pelamis carcass). Change in the numbers of S. kaupii was hence dependent on the arrival of a more powerful scavenger throughout the study site, and at the deeper stations where the population of C. coelolepis declined, S. kaupii was observed to be present but waited for >2 h before feeding, thus contradicting conventional scavenging assumptions in the presence of a food fall.     

Proximate composition, fatty acid and lipid class composition of the muscle from deep-sea teleosts and elasmobranchs

Proximate composition of muscle was determined for the following deep-sea fish species: roughhead grenadier (Macrourus berglax), mora/deep-sea cod (Mora moro), Portuguese dogfish (Centroscymnus coelolepis), black dogfish (Centroscyllium fabricii), leafscale gulper shark (Centrophorus squamosus), greater lantern shark (Etmopterus princeps), smalleyed rabbitfish/ghostshark (Hydrolagus affinis), birdbeak dogfish (Deania calcea) and two species of smooth head (Alepocephalus bairdii and Alepocephalus agassizii). The first eight species contained less than 1% fat in the muscle, while the last two contained 3.0% and 3.6% fat, respectively. Fatty acid and lipid class composition was determined for the first five fish species and showed that the dominant class of lipids was phospholipids. The lipids consisted mainly of polyunsaturated fatty acids (PUFA), and docosahexaenoic acid (DHA) was the dominant fatty acid. Roughhead grenadier and mora showed resemblance to cod (Gadus morhua) regarding protein content, fat content and fatty acid composition. However, the muscle from the deep-sea fish species did contain a higher proportion of arachidonic acid (20:4n-6) than cod muscle.     

Purification and characterization of a Neu5Acalpha2-6Galbeta1-4Glc/GlcNAc-specific lectin from the fruiting body of the polypore mushroom Polyporus squamosus

A lectin has been purified from the carpophores of the mushroom Polyporus squamosus by a combination of affinity chromatography on beta-D-galactosyl-Synsorb and ion-exchange chromatography on DEAE-Sephacel. Gel filtration chromatography, SDS-polyacrylamide gel electrophoresis, and N-terminal amino acid sequencing indicated that the native lectin, designated P. squamosus agglutinin, is composed of two identical 28-kDa subunits associated by noncovalent bonds. P. squamosus agglutinin agglutinated human A, B, and O and rabbit red blood cells but precipitated only with human alpha(2)-macroglobulin, of many glycoproteins and polysaccharides tested. The detailed carbohydrate binding properties of the purified lectin were elucidated using three different approaches, i.e. precipitation inhibition assay (in solution binding assay), fluorescence quenching studies, and glycolipid binding by lectin staining on high-performance thin layer chromatography (solid-phase binding assay). Based on the results obtained by these assays, we conclude that although the P. squamosus lectin binds beta-D-galactosides, it has an extended carbohydrate-combining site that exhibits highest specificity and affinity toward nonreducing terminal Neu5Acalpha2, 6Galbeta1,4Glc/GlcNAc (6'-sialylated type II chain) of N-glycans (2000-fold stronger than toward galactose). The strict specificity of the lectin for alpha2,6-linked sialic acid renders this lectin a valuable tool for glycobiological studies in biomedical and cancer research.     

ZONE LINES IN PLANT TISSUES

The black lines of Polyporus squamosus have been found in the wood of elms. Isolations were made from these lines and an account of the development of P. squamosus in artificial culture is given. Particular mention is made of the formation of black plates, or lines as they appear in section, in culture media and of the appearance of abnormal fructifications. Pure cultures of the fungus on sterilized wood blocks have produced black lines in the wood, similar to those occurring in nature.
From a consideration of the structure and formation of the black plates, the suggestion is made that they form the limiting layer or rind of sclerotium-like bodies buried in the attacked wood.
A review is given of sclerotium formation and structure in several species of Polyporus, and analogies are made between these sclerotia and the sclerotium-like body of P. squamosus. It is decided to distinguish the P. squamosus body from these sclerotia by naming it a pseudo-sclerotium, on the criteria previously advanced for Armillaria mellea (8).     

Molecular diversity of marine glues: polyphenolic proteins from five mussel species.

Adhesive polyphenolic proteins have been purified and characterized from the feet of five marine mussels (Brachidontes exustus, Modiolus modiolus squamosus, Mytella guyanensis, Septifer bifurcatus, and Trichomya hirsuta). All five proteins contain high levels of 3,4-dihydroxyphenylalanine (DOPA), lysine, glycine, and serine or threonine. All but B. exustus also contain high levels (> or = 10%) of proline or 4-hydroxyproline. The polyphenolic proteins of all the mussels have repeated sequences of the motif X1-Y*-X2-Y*-X3-K, where Y* denotes tyrosine or DOPA. In two species (S. bifurcatus and B. exustus), X2 represents 3 amino acids (frequently glycine) and X3 is absent. M. guyanensis is similar except that X2 is reduced to 2 amino acids. In T. hirsuta and M. m. squamosus, however, X2 is absent and X3 occurs as alanine or hydroxyproline. All proteins share approximately equimolar proportions of tyrosyl- and lysyl-derived residues. Although all of the mussels examined thus far are adhesively opportunistic with respect to substratum type, a rigidly invariant sequence does not appear to be necessary for achieving this.     

Pupal development of Ceratitis capitata (Diptera: Tephritidae) and Diachasmimorpha longicaudata (Hymenoptera: Braconidae) at different moisture values in four soil types

This study aimed to evaluate adult emergence and duration of the pupal stage of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann), and emergence of the fruit fly parasitoid, Diachasmimorpha longicaudata (Ashmead), under different moisture conditions in four soil types, using soil water matric potential. Pupal stage duration in C. capitata was influenced differently for males and females. In females, only soil type affected pupal stage duration, which was longer in a clay soil. In males, pupal stage duration was individually influenced by moisture and soil type, with a reduction in pupal stage duration in a heavy clay soil and in a sandy clay, with longer duration in the clay soil. As matric potential decreased, duration of the pupal stage of C. capitata males increased, regardless of soil type. C. capitata emergence was affected by moisture, regardless of soil type, and was higher in drier soils. The emergence of D. longicaudata adults was individually influenced by soil type and moisture factors, and the number of emerged D. longicaudata adults was three times higher in sandy loam and lower in a heavy clay soil. Always, the number of emerged adults was higher at higher moisture conditions. C. capitata and D. longicaudata pupal development was affected by moisture and soil type, which may facilitate pest sampling and allow release areas for the parasitoid to be defined under field conditions.     

Age estimation of the exploited deepwater shark Centrophorus squamosus from the continental slopes of the Rockall Trough and Porcupine Bank

Dorsal spine sections of the deepwater squalid shark Centrophorus squamosus provided age estimates of 21–70 years. Small specimens were not recorded in the study area. It was not possible to obtain estimates from vertebral centra. The estimates are discussed in the context of other studies using dorsal spines of squalid sharks. Sexual maturity was achieved at large size, >75% of maximum length. Total length at 50% maturity was calculated as 101 cm (males) and 128 cm (females).     

New records for Dinemoura ferox (Copepoda: Siphonostomatoida: Pandaridae) from Pacific sleeper sharks captured in waters off eastern Taiwan

Four of the 27 Pacific sleeper sharks (Somniosus pacificus Bigelow and Schroeder) captured in the western North Pacific Ocean off eastern Taiwan between 19 March and 18 May 2002 hosted the parasitic copepod Dinemoura ferox (Kr?yer, 1838) on their body surface including the fins. This report documents a new host record as well as a new ocean record for D. ferox, which until now has only been reported from the benthopelagic sharks, Somniosus microcephalus (Bloch and Schneider) and Centrophorus squamosus (Bonnaterre), occurring in the north Atlantic Ocean off Greenland and Iceland.     

Characterization of neutral and acidic glycosphingolipids from the lectin-producing mushroom, Polyporus squamosus

The polypore mushroom Polyporus squamosus is the source of a lectin that exhibits a general affinity for terminal beta-galactosides, but appears to have an extended carbohydrate-binding site with high affinity and strict specificity for the nonreducing terminal trisaccharide sequence NeuAcalpha2 --> 6Galbeta1 --> 4Glc/GlcNAc. In considering the possibility that the lectin's in vivo function could involve interaction with an endogenous glycoconjugate, it would clearly be helpful to identify candidate ligands among various classes of carbohydrate-containing materials expressed by P. squamosus. Since evidence has been accumulating that glycosphingolipids (GSLs) may serve as key ligands for some endogenous lectins in animal species, possible similar roles for fungal GSLs could be considered. For this study, total lipids were extracted from mature fruiting body of P. squamosus. Multistep fractionation yielded a major monohexosylceramide (CMH) component and three major glycosylinositol phosphorylceramides (GIPCs) from the neutral and acidic lipids, respectively. These were characterized by a variety of techniques as required, including one- and two-dimensional (1)H- and (13)C-nuclear magnetic resonance (NMR) spectroscopy; electrospray ionization-mass spectrometry (ESI-MS, tandem-MS/collision-induced decay-MS, and ion trap-MS(n)); and component and methylation linkage analysis by gas chromatography-mass spectrometry. The CMH was determined to be glucosylceramide having a typical ceramide consisting of 2-hydroxy fatty-N-acylated (4E,8E)-9-methyl-sphinga-4,8-dienine. The GIPCs were identified as Manalpha1 --> 2Ins1-P-1Cer (Ps-1), Galbeta1 --> 6Manalpha1 --> 2Ins1-P-1Cer (Ps-2), and Manalpha1 --> 3Fucalpha1 --> 2Galalpha1 --> 6Galbeta1 --> 6Manalpha1 -->2Ins1-P-1Cer (Ps-5), respectively (where Ins = myo-inositol, P = phosphodiester, and Cer = ceramide consisting mainly of long-chain 2-hydroxy and 2,3-dihydroxy fatty-N-acylated 4-hydroxy-sphinganines). Of these GSLs, Ps-2 could potentially interact with P. squamosus lectin, and further investigations will focus on determining the binding affinity, if any, of the lectin for the GIPCs isolated from this fungus.     

The relation between toxicity and toxin-related-antigen contents of Clostridium botulinum types C and D cultures as determined by mouse bioassay and ELISA

ELISA was tested for the adequacy to differentiate between botulinum type C1 and D toxins. The results presented in this paper indicate antigenic relationship between type C1 and D toxins. Furthermore, indications were obtained that the antigenicities of type C1 and of D toxins produced by different strains are not identical. Although ELISA can be used to differentiate type C1 and D toxins from those of other types, the assay has only limited value to differentiate between type C1 and D toxins. No parallel relationship between toxicity (mouse bioassay) and toxin-related-antigen contents (ELISA) of C. botulinum type C and D cultures was found.     

Changes in monogenean and copepod infestation on captive Acanthopagrus australis (Sparidae)

Infection levels by 17 species of ectoparasite on 491 yellowfin bream collected throughout 1990 from Moreton Bay, S.E. Queensland were compared to infections on 82 bream collected during the same period from a captive population in a large pond at Sea World, Gold Coast, Queensland. There was a significant increase in prevalence and/or intensity of monogeneans Lamellodiscus acanthopagri and Polylabroides multispinosus , but a decrease in the monogenean Anoplodiscus australis and the copepods Ergasilus australiensis, Lernanthropus atrox, Bomolochus stocki and Alella macrotrachelus on captive fish. Twenty-eight bream collected from the pond during autumn were placed in an experimental 1 m³ cage within the pond for 4–6 weeks. Compared to the baseline data for the pond, the caged fish showed increased prevalences of the monogeneans Lamellodiscus acanthopagri, L. squamosus, L. major and Haliotrema spariensis , and increased intensities of L. squamosus, Allomurraytrema robustum and P. multispinosus. The copepod Caligus epidemicus dropped off in preservative, but its abundance (average number per fish) was significantly higher on caged fish than on pond or wild fish. Increased infection levels by L. acanthopagri and A. robustum were due in part to autoinfection. The large skin area promoted large populations of C. epidemicus.     

Comparison of Clostridium botulinum toxins type D and C1 in molecular property, antigenicity and binding ability to rat-brain synaptosomes

Botulinum type D neurotoxin was purified 950-fold from the culture supernatant with an overall yield of 32%. The purified toxin had a specific toxicity of 5.8 X 10(7) mouse minimal lethal dose per mg of protein and a relative molecular mass of 140000. The purified toxin had a di-chain structure consisting of heavy and light chains with relative molecular masses of 85000 and 55000, respectively, linked by one disulfide bond. These subunits had different amino acid compositions and antigenicities. A similarity in molecular constructions and amino acid compositions was observed between type D and type C1 toxins as well as between their subunits. Among the seven kinds of monoclonal antibodies against type D toxin, six reacted with the heavy chain of type D toxin, while one of the six also reacted with the heavy chain of type C1 toxin and neutralized the toxicities of the two toxins. The other one of monoclonal antibodies reacted with the light chains of both toxins. This evidence indicates that both toxins have common antigenic sites on their heavy and light chains and that the antigenic site on the heavy chain may contribute to the neutralization of both toxins by antibody. The binding of type D toxin to rat brain synaptosomes was examined by use of 125I-labelled type D toxin. The binding was competitively inhibited not only by unlabelled type D and C1 toxins, but also by the heavy chains of both toxins, however, it was not inhibited by the light chain of type D toxin. These results suggest that the toxin receptors on synaptosomal membrane are common for type D and C1 toxins, and that the heavy chain contributes to the binding of toxin to synaptosomes and the structure of the binding sites on the heavy chains of both toxins is quite similar.     

Identification of Isolates of Clostridium perfringens Types C and D by Agglutination and Fluorescent-Antibody Methods

Agglutination and fluorescent-antibody methods were employed for screening Clostridium perfringens types C and D from 393 isolates of this organism. All of 50 strains which were isolated in Japan and were agglutinable with an antiserum prepared against a stock strain of type C no. 3182 toxigenically belonged to type C, but the antiserum showed no cross-agglutination with any of type C strains isolated in Denmark. All of the latter strains, however, were agglutinated by an antiserum prepared against a Danish strain, CWC11. Of 64 strains, showing heat-labile agglutinability by type D antiserum L9, 22 strains were toxigenically identified as type D strains which can be divided into three groups by the heat-stable antigens; no strains which were L-agglutination-positive but O-agglutination-negative were epislon-toxigenic. All of 13 strains, the heat-stable antigen of which was agglutinable by a type D antiserum VX81, were toxigenically type D strains. The results of fluorescent-antibody tests were almost in agreement with those of agglutination test with type C strains and completely with those of the O-agglutination test with type D strains. No beta-, epsilon- or delta-toxigenicity could be demonstrated in strains which were not agglutinated by our test sera for types C and D strains. Further examination of cultural properties of Japanese and Danish type C strains revealed that the two groups were considerably different in urease production, capsule formation, and delta- and alpha-toxigenicities.     

Variation des protéins de l'hémolymphe pendant le cycle de mue,chez Penaeus kerathurus

The variation of hemolymph proteins during the moulting cycle of P. thurus is presented, as well as the variation of the total content of water. The concentration of protein is highly variable, and these are important differences according to the sex and the moulting stage. Forty six protein fractions whose molecular weight ranged from 58000 to 1475000 daltons were identified. In the female, the number of protein fractions was as follows: 38 in stage B, 42 in stage C and 26 in stage D. In the male, the electrophoregram is less diverse. Absence of the fractions that migrate at the level of the lipoproteins is obvious. These lipoproteins, however, are a distinctive feature found in the female specimens studied. The variation of the total water content at the moment of moulting is in the order of 4%, with a maximum of 74.5 in stage B₂ and a minimum of 70.35 in stage D₁D₂. The water/dry weight ratio is negative and not significant in stages B and D; and positive and not significant in stage C. The water/wet weight ratio is also negative and not significant in stages B and D, and positive and not significant in stage C.     

Heteromorphism of the corynebacteria. III. D- (dark) and C- (clear) cell types and multiseptate specimens]

A population of Corynebacterium diphtheriae may consist of cells with accented (osmiophilic) cytoplasm (cells of the "dark" type, or D type) and cells with cytoplasm having no pronounced osmiophilic properties (cells of the "clear" type, or C type). The divergence of the population into cells of the D and C types occurs at the stage of cell division, the original mother cell being able to divide into 2 or more individual cells belonging to different types (elongated multiseptate cells). At the same time no morphological disturbances in septation may be observed. The ability of each type of cells for division into the corresponding individual daughter cells indicates their being biologically valid. The mixed (D--C) population of Corynebacterium diphtheriae is considered to be a sign of the dissociation of bacterial culture.     

Description of two new species of Bassozetus (Ophidiiformes: Ophidiidae) and a redescription of Bassozetus robustus Smith and Radcliffe 1913

Ichthyological Research - Two new deepwater assfish species, Bassozetus trachibranchus and Bassozetus squamosus, are described from the 29 western Atlantic and four western Indian and western South...     

Interconversion of Type C and D Strains of Clostridium botulinum by Specific Bacteriophages

These studies show that Clostridium botulinum types C and D cultures can be cured of their prophages and converted to either type C or D depending on the specific phage used. Strains of types C and D were cured of their prophages and simultaneously ceased to produce their dominant toxins designated as C(1) and D, respectively. Cured nontoxigenic cultures derived from type C strain 162 were sensitive to the phages from the toxigenic type C strain 162 and type D strain South African. When cured nontoxigenic cultures derived from strain 162 were infected with the tox(+) phages from the 162 strain of type C and the South African strain of type D, they then produced toxin neutralized by types C and D antisera, respectively. Cured nontoxigenic cultures isolated from the type D South African strain were only sensitive to the parent phage, and, when reinfected with the tox(+) phage, they produced toxin neutralized by type D antiserum. Type C strain 153 and type D strain 1873, when cured of their respective prophages, also ceased to produce toxins C(1) and D, but, unlike strain 162 and the South African strain, they continued to produce a toxin designated as C(2). When the cured cultures from strains 153 and 1873 were infected with the tox(+) phage from type D strain 1873, the cultures simultaneously produced toxin that was neutralized by type D antiserum. When these cured cultures were infected with the tox(+) phage from type C strain 153, the cultures produced toxin that was neutralized by type C antiserum. These studies with the four strains of C. botulinum confirm that the toxigenicity of types C and D strains requires the continued participation of tox(+) phages. Evidence is presented that types C and D cultures may arise from a common nontoxigenic strain.     

Immuno-crossreactivity between botulinum neurotoxin type C1 or D and exoenzyme C3

Botulinum neurotoxin type D and exoenzyme C3 have been separately purified from Clostridium botulinum strain D-1873 to apparent homogeneity. Both ADP-ribosylated a rat liver cytosolic protein of 24 kDa. The N-terminal amino acid sequence of C3 was determined and showed a low degree of homology with those of the light and heavy chains of neurotoxins of various types which have been reported previously. However, a polyclonal antibody raised against C3 cross-reacted with the light chains, but not with the heavy chains, of type C1 and D neurotoxins. Furthermore, a monoclonal antibody recognizing the light chains of type C1 and D neurotoxins interacted with C3. These results suggest that the light chain of type C1 or D neurotoxin and exoenzyme C3 share at least one epitope in common with each other.