Rapid Assessment of Maturation Stage and Reproductive Mode in Centrolecytic Eggs of Stick Insects (Phasmatodea) Using DAPI Stain

A rapid three-step DAPI technique is proposed for detecting meiotic stages and sperm head evolution in yolky, fertilized stick insect eggs, which were difficult to analyze with other methods. Fixed eggs were freed from chorionic envelopes and stained directly in DAPI/PBS solution. After rinsing, eggs were singly squashed in a drop of mounting buffer and examined under a microscope with incident fluorescent illumination. The method was almost uniformly successful, and direct observation of nuclear structures, coupled with fluorometry, allowed easy recognition of bivalents, diads, pronuclei and their DNA content. The DAPI method proposed here appears particularly helpful for investigating unusual reproductive modes in eggs with large amounts of yolk.     

Biotechnology and fish culture

Biotechnology can currently be considered of importance in aquaculture. The increase in the production of aquatic organisms over the last two decades through the use of biotechnology indicates that in a few generations biotechnology may overtake conventional techniques, at least for the commercially more valuable species. In the last few years, genetics has contributed greatly to fish culture through the application of the more recent techniques developed in biotechnology and in genetic engineering. At present, the most commonly used methods in fish biotechnology are chromosome manipulation and hormonal treatments, which can be used to produce triploid, tetraploid, haploid, gynogenetic and androgenetic fish. These result in the production of individuals and lineages of sterile, monosex or highly endogamic fish. The use of such strategies in fish culture has as a practical objective the control of precocious sexual maturation in certain species; other uses are the production of larger specimens by control of the reproductive process and the attainment of monosex lines containing only those individuals of greater commercial value. The use of new technologies, such as those involved in gene transfer in many species, can result in modified individuals of great interest to aquaculturists and play important roles in specific programmes of fish production in the near future.     

Characterization and fine-structural localization of actin-and fibronectin-like proteins in planaria (Dugesia lugubris s.l.)

Summary Actin- and fibronectin-like proteins were characterized in the planarian, Dugesia lugubris s.l., by sodium dodecyl sulphate polyacrylamide gel electrophoresis and immunoblotting analysis using antisera to vertebrate actin and fibronectin. These antisera recognized protein bands of 42 kDa and 220 kDa, respectively. In addition, the immunohistochemical distribution of both actin- and fibronectin-like material was examined by using immuno-electron microscopy. Actin-like protein was localized in myofibrils in various differentiation stages, and in the peripheral cytoplasm and lamellipodia of cells that were migrating. The fibronectin-like component was associated with the extracellular matrix in the fibrillar structures and with the surface of the migrating cells. Our data suggest that similar cellular and molecular mechanisms are involved in cell-matrix interactions and in the morphogenesis of living organisms at different evolutionary levels.     

The mammalian anti-α-smooth muscle actin monoclonal antibody recognizes an α-actin-like protein in planaria (Dugesia lugubris s.l.)

The presence of an alpha-smooth muscle (alpha-sm) actin-like protein in planaria (Dugesia lugubris s.l.) is reported. The protein shows a 42 kDa molecular weight determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis and is specifically recognized by the mammalian anti alpha-sm actin monoclonal antibody. When a planarian is induced to regenerate by head amputation, the immunostaining of the alpha-sm actin-like molecule becomes important in the area of growing blastema, reaching a maximum between 70-120 hours after injury. Conventional electron microscopy at the 4-day-regeneration stage shows that blastema-forming cells are a homogeneous population whose morphological features resemble those of migrating mesenchyme-like cells; only the myoblasts show a recognizable phenotype. The immunocytochemical localization of alpha-sm actin-like molecule by immunoperoxidase (light microscopy) and immunogold stains (electron microscopy) was carried out on both intact and injured worms. The antigen was localized mainly at the basal portion of the epidermal cells and in the undifferentiated mesenchyme-like cells. Myoblasts, but not differentiated myofibers, were also labelled by this antibody. The results indicate that in the lower Eumetazoan planarians, as well as in vertebrates, the alpha-sm actin can be considered to be a marker for myoid differentiation. The suggestion that alpha-sm actin can be used as a marker for mesenchyme-like cells in vertebrates and in invertebrates is also discussed.     

Variation of reproductive effort between parthenogenetic and sexual populations of the scorpion Tityus columbianus

Abstract. Geographic differences in reproductive effort were examined between sexual and parthenogenetic populations of the Colombian scorpion Tityus columbianus , from Iza (sexual) and from Mosquera (unisexual). The sexual females from Iza were significantly larger and had significantly greater relative litter mass (RLM) than the parthenogenetic ones from Mosquera. In both populations litter size increased significantly with female body size.
Litter size and RLM in the sexuals averaged 12.7 and 0.49 respectively, and in the unisexuals 8 and 0.36. We suggest that the higher values are related to greater primary productivity at the Iza site.     

Relationship between thermal behaviour,fermentation performance and fatty acid composition in two strains of Saccharomyces cerevisiae

Two Saccharomyces cerevisiae strains, one specifically of S. cerevisiae and another belonging to the physiological race S. uvarum, exhibited associative and dissociate thermal profiles, respectively. The S. cerevisiae subsp. uvarum strain, which displayed the dissociative profile, was characterized by a higher aptitude for fermenting glucose in a superoptimal temperature range as well as by a lower fatty acid unsaturation degree. On the other hand, both strains exhibited a similar fatty acid composition modulation pattern with regard to temperature: the unsaturation level presented two relative maxima at 15 and 40° C. However, on the basis of Central Composite Design results, supplementation with an oleic acid source under semi-anaerobic conditions did not improve the fermentative performances in either strain. the modelling of fermentation rate in relation to certain variables indicated that the fermentative performance at superoptimal temperatures, and particularly the optimal temperature (T _opt) and maximal temperature (T _max) of the strain displaying a dissociative profile, could be increased by acting on medium composition. Correspondence to: M. E. Guerzoni