Triploidy induction in Heterobranchus longifilis (Family: Clariidae) by cold shock

Triploid was induced in African Catfish (Heterobranchus longifilis) by cold shocking activated eggs at 5 degree C for forty minutes starting 3-4 minutes after fertilization. Triploidy was confirmed from mitotic chromosomes prepared from embryo which showed 100% triploidy in the cold shocking treatment and 100% diploidy in the control treatment

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.

Induced diploid meiogynogenesis in the African clariid catfish Heterobranchus longifilis (Valenciennes, 1840) (Pisces: Clariidae)

Diploid meiotic gynogenesis was induced in African catfish, Heterobranchus longifilis by injection of 0.5ml/kg ovaprim on the breeders, followed by application of UV light irradiation on the spermatozoa and temperature shocking of activated eggs. Diploidy was restored by shocking haploid activated eggs at 5 degree C for 40 minutes. The normal control spermatozoa did not receive any UV irradiation nor temperature shock, while the haploid control spermatozoa were irradiated, but did not receive cold shock. The percentage hatchability in the treated group was 25%, while in the control it was 53%. Less than 15 fingerlings had morphological aberrations. After two weeks of indoor rearing, the survival percentage of the treated group was 45% in the control experiment. Cytogenetic analysis of chromosomes revealed 25 chromosomes in the haploid embryo and 50 chromosomes each in diploid gynogenesis and normal diploid control

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

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 comparative study on the embryonic development of gynogen, triploid, haploid and normal diploid embryos of stinging catfish, Heteropneustes fossilis

UV irradiation and cold shock were applied on the eggs of stinging catfish, Heteropneustes fossilis, to produce haploid,. gynogen and triploid embryos. A comparative account of the various features· of embryonic development in chromosomally manipulated groups viz. haploid, gynogen and triploid and non-manipulated normal diploid group of H fossilis has been discussed. A slow development and delayed hatching were observed in gynogen and triploid embryos compared to those in normal diploid (control) groups. Mass mortality was observed in all chromosomally manipulated groups particularly during the gastrulation stage. The hatchlings of the gynogen, triploid and normal diploid were similar in overall appearance.

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.

Influence of cold storage time on the protein changes and fatty acids deterioration of (Rutilus frisi kutum) during frozen storage

The main aim of this research was to identify fatty acids composition of Caspian sea of White fish Rutilus frisi kutum tissue and their changes during one year cold storage (-18Ċ).The secondary aim was to determine the changes of moisture, ash, protein, fat, and to investigate the effects of storage time on peroxide, TBAi, FFA, and extractability of myofibrillar proteins of the fish tissue during one year cold storage (-18 Ċ). 10 samples of (Rutilus frisi kutum) were randomly collected from Anzali landings. The samples were frozen at -30 Ċ and kept in cold storage at -18Ċ for one year. According to time table, the samples were examined. The results showed that 27 fatty acids were identified. The unsaturated fatty acids (UFA) and saturated fatty acids (SFA) were 74/09 and 21/63 %, respectively, in fresh tissue. So that DHA (C22:6) oleic acid (C18:1c) had high amounts (15/07 ,20/57 ) among the UFA and palmitic acid (C16:0) was the most (13/09 %) among the SFA. The effects of freezing and cold storage on fish tissue showed that UFA and SFA contents have reached to 58/79 and 22/17 %, respectively, at the end of cold storage. It indicated that these compound change to each other during frozen storage. Also ω-3 and ω-6 series of fatty acids was 24/22 and 15/56% in fresh tissue, but their contents decreased to 8/68 and 5/11% at the end of period. Among the fatty acids C22:6, C18:1c and C16:0 had the most changes. The changes of fatty acids were significantly at 95% level expected for C18:0. Results showed that moisture, ash, protein, and fat contents were 75/9±0/03, 1/28±0/012, 21/8±0/2, and 4/1±0/01 % respectively, in fresh tissue. The moisture, ash, protein, and fat contents were 72/3±0/04, 1/83±0/05, 1/91±0/01 and 19/9±0/01 % respectively, at the end of storage period. Lipid damage was measured on the basis of free fatty acids (FFA), peroxide value (PV), and Thiobarbituric acid index (TBA-i). PV, TBARS and FFA concentration of frozen Caspian Sea white fish stored at -18 Ċ the temporal variation of these three variables were statistically significant (p<0.001). Results of White fish myofibrillar proteins showed aggregation of bound reduced for stored at 12 months. SDS-PAGE analysis revealed that, the intensity of the myosin heavy chain and actin bound was reduced with increasing storage time. SDS-PAGE patterns showed that myosin heavy chain was much more susceptible to hydrolysis than actin. Key words: Rutilus frisi kutum, frozen storage, ω-3, ω-6, protein myofibrillar

Application of whey protein incorporation with Cinnamon oil as a preservative in Huse huso fillet under chilled storage condition

Biodegradable protein-based film was developed by incorporating cinnamon essential oil (CEO) into whey protein concentrate (WPC) at level of 0.8% and 1.5% v/v. Then physical and mechanical properties of the films were evaluated. Adding CEO to the WPC matrix decreased the water vapour permeability of the films and water solubility. Films containing CEO showed significant antibacterial activity both gram-positive and gram-negative strains and exhibited significant inhibitory effect on the studied fungi. In continue, the effect of whey coating and whey coating incorporated with 1.5% CEO on quality and shelf life of Huso huso fillet during refregrated (4±1°C) storage period were also investigated. The control and treated fish samples were analyzed for microbiological (total viable count, psychrophilic counts), chemical (PV, TBA, FFA, pH, TVB-N), and sensory characteristics in 4-day intervals up of microbial, chmical and sensoy analyses indicated lower levels of PV, TBA, FFA, pH, TVB-N in coasted sampels and specially, those with CEO while were kept in refrigerator. Based on results, whey protein edible coating contain 1.5% cinnamon essential oil could enhance preserving ability Huso huso during storage cold.

Identification and measurement of fatty acids, amino acids in roe of tuna fish (Thunnus tonggol) and its TVB and peroxide value during cold storage

The first aim of this research was to identify fatty acids, amino acids composition of Thunnus tonggol roe and their changes during cold storage (-18'C). The second aim was to determine the changes of moisture, protein, fat and ash contents of the roe during one year cold storage (-18'C). 60 samples of longtail tuna (Thunnus tonggol) ovaries were randomly collected form Bandar-e-Abbas landings. The samples were frozen at-30'C and kept in cold store at -18'C for one year. According to a time table, the samples were examined for identification of fatty acids, amino acids, moisture, protein, fat, ash, peroxide and T.V.N. and their changes were evaluated during this time. The results showed that 26 fatty acids were identified. The unsaturated fatty acids (UFA) and saturated fatty acids (SFA) were 62.33 and 37.6%, respectively, in fresh roe. So that, DHA (C22:6) and oleic acid (C18:1) had high amounts (24.79 and 21.88%) among the UFA and palmitic acid (C16:0) was the most content (22.75%) among the SFA. The PUFA/SFA was 0.91. Also, 17 amino acids were identified that essential amino acids (EAA) and nonessential amino acids (NE) were 10478 and 7562 mg/100g, respectively, and E/NE was 1.38. Among the EAA and NE, lysine (2110mg/100g) and aspartic acid (1924 mg/100g) were the most contents. Also, results showed that moisture, ash, protein and fat contents were 72.74, 1.8, 19.88 and 4.53%, respectively, in fresh roe. The effects of freezing and cold storage on the roes showed that UFA and SFA contents have reached to 49.83 and 48.07%, respectively, at the end of cold storage. It indicated that these compounds change to each other during frozen storage. Also, n-3 and n-6 series of fatty acids were 32.75 and 1.61% in fresh roe. But their contents decreased to 22.96 and 1.25% at the end of period. Among the fatty acids, 22:6 and C16:0 had the most changes. The changes of fatty acids were significantly at 95% level except for C15:1, C18:3(n-3) and C20:4(n-6). All of the amino acids decreased in frozen storage and their changes were significantly (P<0.05). EAA was 7818 mg/100g and E/NE was 1.27 at the end of storage period. Among the amino acids, leucine and lysine had the most changes. Moisture, ash, protein and fat contents were 70.13, 1.82, 19.4 and 6.51%, respectively, at the end of storage period. The peroxide value and T.V.N. increased during storage. So that, their contents have reached to 5.86 mg/kg and 26.37 mg/100 g, respectively, at the end of frozen storage. The best shelf life of Thunnus tonggol roe was 6 or 7 months, because of lipid oxidation and increasing of peroxide.