Molecular studies on the effect of germanium oxide on sponges, corals and ascidians

The stress response, at the molecular level, of the soft corals Dendronephthya klunzingeri and Heteroxenia sp., hard corals Acropora hyacinthus and A. valenciennesi, an ascidian Symplegma sp. and sponges Latruncula cortica and Callyspongia crassa to germanium oxide (GeO sub(2)) was evaluated. Evaluation was carried out using bioindicators. such as the level of expression of each of the heat shock proteins (HSPs) and the silicatein enzyme in response to the compound. However, the expression was measured by SDS Polyacrylamide Gel Electrophoresis (SDS PAGE) and western blotting. The harmful concentration of GeO sub(2) that produced noticeable molecular changes in the studied samples during the first 6-24 hours was 6 μg/ml. The two studied soft corals as well as the ascidian responded to the harmful concentration of germanium oxide by expressing the heat-shock protein 90 (hsp90), while the two hard corals responded by expressing hsp70, C. crassa by decreasing the level of silicatein enzyme and sponge L. cortica produced no change by any of the used biomarkers, The soft coral Heteroxenia sp. was found to be sensitive to mechanical stress during the experiment and it was more sensitive to 6 μg/ml of GeO sub(2) than the other soft coral D. klunzingeri. The two studied hard corals were sensitive to mechanical stress during the experiment, but A. hyacinth us showed higher sensitivity than A. valenciennesi. However, these 2 corals displayed reverse response to GeO sub(2). Primitive evidences were found in the SDS PAGE to distinguish the tissue of the soft coral from that of the hard coral on the molecular level; the soft coral showed two prominent protein bands (45 and 50 kDa) while the two prominent protein bands for hard corals were 31 and 116 kDa.

Heat-stress and light-stress induce different cellular pathologies in the symbiotic dinoflagellate during coral bleaching

Coral bleaching is a significant contributor to the worldwide degradation of coral reefs and is indicative of the termination of symbiosis between the coral host and its symbiotic algae (dinoflagellate; Symbiodinium sp. complex), usually by expulsion or xenophagy (symbiophagy) of its dinoflagellates. Herein, we provide evidence that during the earliest stages of environmentally induced bleaching, heat stress and light stress generate distinctly different pathomorphological changes in the chloroplasts, while a combined heat- and light-stress exposure induces both pathomorphologies; suggesting that these stressors act on the dinoflagellate by different mechanisms. Within the first 48 hours of a heat stress (32°C) under low-light conditions, heat stress induced decomposition of thylakoid structures before observation of extensive oxidative damage; thus it is the disorganization of the thylakoids that creates the conditions allowing photo-oxidative-stress. Conversely, during the first 48 hours of a light stress (2007 µmoles m−2 s−1 PAR) at 25°C, condensation or fusion of multiple thylakoid lamellae occurred coincidently with levels of oxidative damage products, implying that photo-oxidative stress causes the structural membrane damage within the chloroplasts. Exposure to combined heat- and light-stresses induced both pathomorphologies, confirming that these stressors acted on the dinoflagellate via different mechanisms. Within 72 hours of exposure to heat and/or light stresses, homeostatic processes (e.g., heat-shock protein and anti-oxidant enzyme response) were evident in the remaining intact dinoflagellates, regardless of the initiating stressor. Understanding the sequence of events during bleaching when triggered by different environmental stressors is important for predicting both severity and consequences of coral bleaching

Comparison of effectiveness of heat and cold shocks applied in the induction of gynogenesis in Clarias gariepinus (Burchell)

An experiment was conducted to optimize the procedure of gynogenesis in African catfish, Clarias gariepinus by suppressing meiotic and mitotic cell divisions in fertilized eggs. Gynogensis was conducted by fertilizing normal eggs with UV-irradiated sperm followed by either heat or cold shocking Irradiation of spermatozoa was given for a duration of 1 min and the eggs were fertilized in vitro. Cold shock at a temperature of 3± 1°C for a duration of 30 and 60 min and heat shock at a temperature of 39± 1°C for a duration of 1 and 2 min was applied to induce diploidy. Higher percentage of hatching (68.66) was observed for meiotic gynogens at a shock temperature of 39± 1°C for a duration of 1 min, 5 min after fertilization (af). Higher percentage of mitotic gynogenetic induction (15.33) was observed at a temperature shock of 39± 1°C for a duration of 1 min, 30 min af.

Nachweis von Parvalbuminen in Störgeweben

Parvalbumins, calcium-binding, heat-stable proteins of considerable allergenic potential, are occurring in high concentration in light muscle of many fish species. Lack of knowledge about parvalbumins in sturgeon tissues prompted us to study the distribution of this type of proteins in light and dark muscle, as well as in swim bladder and skin of Atlantic sturgeon, Acipenser oxyrhynchus, and three other sturgeon species. Results: Light and dark muscle of sturgeons contained water soluble proteins with following properties: (1) Great stability against heating at 70 or 80 °C. (2) Isolelectric points between 3.85 and 5.66. (3) Treatment of proteins separated by isoelectric focusing with the dye “stains all” gave blue protein bands. (4) The molecular mass was about 10 to 14 kDalton. (5) Concentration of acidic, heat-stable proteins was higher in light than in dark muscle. Taken together, these findings gave strong indication for the presence of parvalbumins in muscle tissue of sturgeons. This conclusion was corroborated by immunological tests. However, parvalbumin could not be detected in swim bladder tissue. Skin preparations showed only traces of parvalbumins, possibly resulting from residual muscle tissue.

Induction of mitotic and meiotic gynogenesis and production of genetic clones in rohu, Labeo rohita Ham.

Studies were undertaken to produce genetic clones derived from all homozygous mitotic gynogenetic individuals in rohu, Labeo rohita Ham. ln view of this, attempts were made to interfere with the normal functioning of the spindle apparatus during the first mitotic cell division of developing eggs using heat shocks, there by leading to the induction of mitotic gynogenetic diploids in the F1 generation. Afterwards, viable mitotic gynogenetic alevins were reared and a selected mature female fish was used to obtain ovulated eggs which were fertilized later with UV-irradiated milt. Milt was diluted with Cortland’s solution and the sperm concentration was maintained at 10⁸/ml. The UV-irradiation was carried out for 2 minutes at the intensity of 200 to 250 µW/cm² at 28± 1°C. The optimal heat shock of 40°C for 2 minutes applied at 25 to 30 minutes a.f. was used to induce mitotic gynogenesis in first (F1) generation and at 3 to 5 minutes a.f. to induce meiotic gynogenesis in the second (F2) generation. The results obtained are presented and the light they shed on the timing of the mitotic and meiotic cell division in this species is discussed.

Estimation of ploidy and hybridization status by erythrocyte and nucleus size analysis in carps

The present experiment was designed to observe whether the nuclear volume and area are affected by the ploidy and hybrid status of the individual. Polyploidy was induced by heat shock treatment given at 44 ± 0.5°C for 30 seconds and 45 seconds which was found to be most effective (64.7%) for induction of triploidy in Cyprinus carpio. Cell and nuclear volume and cell and nuclear area varied significantly in triploid fishes as compared to those of controls. Triploid fishes showed significantly higher growth compared to diploid counterparts. It was also observed that catla x rohu hybrid and its parents showed significant difference in the nuclear volume and area of their erythrocytes. Except nuclear volume, all the parameters were significantly different between catla and catla x rohu hybrid. The hybrids showed a closer relationship with catla as compared to rohu.

Production of triploid African catfish, Clarias gariepinus (Burchell), using chromosome manipulation techniques

An experiment was conducted to induce triploidy in African catfish, Clarias gariepinus, using heat shock and cold shock techniques. Cold shock at a temperature of 0± 1°C and 5±1°C for a duration of 15, 30, 45 and 60 min and heat shock at a temperature of 40±0.5°C and 41 ±OS C for a duration of 1, 2 and 3 min was given to induce triploidy 5 min after fertilization. Maximum percentage of triploids (91.4%) were obtained in the heat shock at a temperature of 40±0SC for a duration of 1 min whereas cold shock at 0± 1 C for a duration of 60 min yielded 90% of triploids. Chromosome analysis revealed that diploids have 54 chromosomes and triploids have 81 chromosomes. The erythrocyte measurements of the minor axis and major axis were 1.17 times larger in treated fish than in controls. The growth studies showed that the growth rate was not significantly affected in triploids.

A study of fish ice cream formulation by use of silver carp proteins instead of milk proteins

Fish is a valuable nutritional source witch use of it in daily meal has a beneficial role on nutritional needs supply and also causes mental and physical health especially in people who have protein and food deficiencies. Unfortunately, per capita consumption of sea foods in Iran is 5.5Kg witch is very lower than world standards. So, study on fish ice cream formulation, by use of fish protein concentrate (FPC) instead of milk protein, had done to make variation in sea foods products and also increase per capita consumption of these kinds of foods. FPC has very high protein concentration and a lot of necessary Also it's protein is very digestible amino acids like lysine and methionine with highly biological value and it's PER in compare with casein PER is high. At first fish protein concentrate type A produced from silver carp in three steps by the extraction with isopropyl alcohol solvent and heat. Microbiological and physicochemical specifications of produced FPC by rules of FDA and FAO were accepted. Finally according to panel test results, substitution of 30 percent of milk with FPC is acceptable. Also microbiological and physicochemical specifications of product were tested and results in compare with national standards of Iran were accepted.