Trichodesmium Ehrenberg species were collected in the Caribbean Sea, Sargasso Sea, and coastal areas of Tanzania (Indian Ocean). The specimens were divided into five species on the basis of morphometric characters such as cell dimensions and colony formation: T. tenue Wille, T. erythraeum Ehrenberg, T. thiebautii Gomont, T. hildebrandtii Gomont, and T. contortum Wille. In addition, Trichodesmium sp., a spherical colony of uncertain taxonomic position was examined. The cell structure of each species was investigated by means of light, scanning electron, and transvnission electron microscopy. Particular attention was paid to the presence and ultrastructural arrangement of gas vacuoles and glycogen fiber clusters (GFCs). This resulted in identification of two major groups of species: 1) T. tenue, Trichodesmium sp. with spherical‐shaped colonies, and T. erythraeum with GFCs and more or less localized gas vacuoles; and 2) T. thiebautii, T. hildebrandtii, and T. contortum lacking GFCs and with gas vacuoles spread at random. The species within each group were further characterized with respect to the dimension of the gas vesicles, cylindrical bodies, scroll bodies, and a new cellular inclusion body, Differences in colony formation and cell dimensions correlated with specific ultrastructural characters in five of the six forms. This is the first ultrastructural study comparing different forms of Trichodesmium sampled at geographically remote areas and shows that one species appears identical regardless of the sampling site. Some of the species had not been investigated earlier, and probably more species are to be identified and analyzed.
Trichodesmium thiebautii Gomont, a marine planktonic diazotrophic cyanobacterium, has an unusual subcellular arrangement. To identify subcellular structures related to photosynthesis, antibodies against phycoerythrin, phycocyanin, and ribulose 1,5‐bisphosphate carboxylase/oxygenase (Rubisco) were used together with an immuno‐gold labeling technique and electron microscopy. Thylakoid membranes, identified by transmission electron microscopy and phycobiliprotein labeling, were arranged as a loose network throughout all cells. Rubisco showed a particularly intense localization in medium electron‐dense polyhedral bodies, therefore identified as carboxysomes. The average density of the carboxysomal Rubisco label was about five times higher than that in the cytoplasm. The carboxysomes (4–11 per cell section) were scattered throughout the cytoplasm. These data, together with those obtained from double immunolabeling experiments using nitrogenase (Fe‐protein) and Rubisco antibodies, revealed that Trichodesmium contains both N2‐ and CO2‐fixing proteins within the same cell. This is in contrast to the previous concept of a spatial segregation of the two processes in Trichodesmium and demonstrate that nitrogenase‐containing cells are not comparable to heterocysts in this context.
Immunochemical labeling was used to study the subcellular distribution of cytochrome oxidase, a respiratory protein, in Trichodesmium thiebautii. The protein was found associated with both cytoplasmic and thylakoid membranes. About a sixfold variation in the protein content (gold particle count) was found among Trichodesmium cells within a single colony. Double labeling was performed with cytochrome oxidase and nitrogenase antisera. Regression analysis of gold particle counts per unit of cell area of cytochrome oxidase and nitrogenase showed a positive correlation (r2 = 0.911); cells with higher nitrogenase levels also had higher levels of cytochrome oxidase. The parallel expression of two proteins suggests that respiratory oxygen uptake may be involved in nitrogenase protection (respiratory protection) in Trichodesmium spp.
We examined freshly collected samples of the colonial planktonic cyanobacterium Trichodesmium thiebautii to determine the pathways of recently fixed N within and among trichomes. High concentrations of glutamate and glutamine were found in colonies. Glutamate and glutamine uptake rates and concentrations in cells were low in the early morning and increased in the late morning to reach maxima near midday; then uptake and concentration again fell to low values. This pattern followed that previously observed for T. thiebautii nitrogenase activity. Our results suggest that recently fixed nitrogen is incorporated into glutamine in the N2-fixing trichomes and may be passed as glutamate to non-N2-fixing trichomes. The high transport rates and concentrations of glutamate may explain the previously observed absence of appreciable uptake of NH4+, N03-, or urea by Trichodesmium spp. Immunolocalization, Western blots (immunoblots), and enzymatic assays indicated that glutamine synthetase (GS) was present in all cells during both day and night. GS appeared to be primarily contained in cells of T. thiebautii rather than in associated bacteria or cyanobacteria. Double immunolabeling showed that cells with nitrogenase (Fe protein) contained levels of the GS protein that were twofold higher than those in cells with little or no nitrogenase. GS activity and the uptake of glutamine and glutamate dramatically decreased in the presence of the GS inhibitor methionine sulfoximine. Since no glutamate dehydrogenase activity was detected in this species, GS appears to be the primary enzyme responsible for NH3 incorporation.
Five benthic species of the genus Gambierdiscus (Dinophyceae) were observed for the first time in the coastal waters of Pakistan, Northern Indian Ocean. The morphology of the epiphytic, ciguatera-related toxic species G. toxicus, G. belizeanus, G. polynesiensis, G. australes and G. cf. yasumotoi are presented here, described by the Kofoid system of thecal plates Po, 3΄, 7˝, 6c, 8s, 5˝΄, 1p, 2˝˝ with differences in cell shape, cell size, plates, pores around the apical pore plate by using light and scanning electron microscopy. The occurrence of these potentially toxic dinoflagellate species in Pakistani coastal areas of Manora Channel and Balochistan during high temperatures of 28-32ºC is cause of concern for human health impacts from ciguatera fish poisoning.
The effects of several dietary protein levels on the growth, feed conversion, body composition and diet digestibility of juvenile Lutjanus argentimaculatus (body weight 12.3 g) were examined. Seven isolipidic (7.4%) diets were formulated to contain graded levels of protein (28–58%) with dietary energy ranging from 19.7 to 21.5 kJ g−1. Diets were distributed to triplicate groups of fish thrice a day at ration of 2% body weight for 90 days. Growth, feed conversion, protein utilization and digestibility of nutrients increased with increasing dietary protein level up to 43%, after which no significant improvement was observed. Digestibility of dry matter and energy showed a concomitant increase with the reduction in dietary wheat meal, attaining maximal values with high protein diets. No significant differences were detected in moisture, protein, lipid and ash content of whole fish or body organs as dietary protein increased. The mesenteric fat, hepato‐ and viscerosomatic indices decreased with increasing protein level. The cholesterol, triglycerides and haematocrit values were similar among treatments, except that high levels of plasma lipids were recorded above 43% protein diet. The use of a practical diet containing 43% protein is appropriate for the growth of L. argentimaculatus juveniles under the experimental conditions of the present study.
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