Characterization of a fibrinogen-clotting enzyme from Trimeresurus stejnegeri venom, and comparative study with other venom proteases

Trimeresurus stejnegeri venom, which contains TSV-PA (a specific plasminogen activator sharing 60-70% sequence homology with venom fibrinogen-clotting enzymes), also possesses fibrinogen-clotting activity in vitro. A fibrinogen-clotting enzyme (stejnobin) has been purified to homogeneity by gel filtration and ion-exchange chromatography on a Mono-Q column. It is a single-chain glycoprotein with a mol. wt of 44,000. The NH2-terminal amino acid sequence of stejnobin shows great homology with venom fibrinogen-clotting enzymes and TSV-PA. Like TSV-PA, stejnobin was able to hydrolyse several chromogenic substrates. Comparative study of substrate specificities of stejnobin and other venom proteases purified in our laboratory was carried out on five chromogenic substrates. Stejnobin clotted human fibrinogen with a specific activity of 122 NIH thrombin-equivalent units/mg protein. However, stejnobin did not act on other blood coagulation factors, such as factor X, prothrombin and plasminogen. Diisopropyl fluorophosphate and phenylmethanesulfonyl fluoride inhibited its activity, whereas ethylenediamine tetracetic acid had no effect on it, indicating that it is a serine protease. Although stejnobin showed strong immunological cross-reaction with polyclonal antibodies raised against TSV-PA, it was interesting to observe that, unlike the case of TSV-PA, these antibodies did not inhibit the amidolytic and fibrinogen-clotting activities of stejnobin. (C) 1998 Elsevier Science Ltd. All rights reserved.

Comparative study on the toxin production and profiles Raphidophytes at different salinities

PLEASE ALSO CHECK THE FULL TEXT ABSTRACT. Toxin production and toxin profiles of four Raphidophytes grown under different salinities were compared to investigate the influence of salinity on cellular content of neurotoxin. In Chatonella andqua CaTx-1, CaTx-11, and CaTx-111 peaked at 25 pplt with yields of 0.99, 0.42, and 2.90 pg/ceU, but the highest yields (2.35 pg/cell) of CaTx-IV was attained at 30 ppt. On the other hand, Chatonella marina yielded higher proportions of CmTx-1 (0.55 pg/ceH) and CmTx-111 (2.50 pg/cell) at 25 ppt. However, CmTx-IV was present in its highest amount (1.65 pg/cell) at 30 ppt, as seen in C anriqua. A smaH amount of CmTx-11 was also detected at 20-35 ppt. The toxin compositions indicate that H. akashiwo is more sensitive to higher salinities than the other three raphidophytes. Substantial compositional change was observed in case of H. akashiwo. HaTx-11 (corresponding to PbTx-9) was detected only as a trace at 20 and 25 ppt. Toxin HaTx-IV (corresponding to oxidized PbTx-2) was most dominant and peaked at 20 ppt with a yield of 0.3 pg/cell. Considerable amounts of HaTx-1 and III (corresponding to PbTx-2 and 3) were also detected. At higher salinities of above 25 ppt HaTx-11 was not detected. F. japonica gave highest yields of FjTx-11 (PbTx-2) and FjTx-IV (Oxidized PbTx-2) at 20 ppt with yields of 0.95, 1.54 pg/cell while the production of toxic profiles FjTx-1 (PbTx- 1) and FjTx-111 (PbTx-3) peaked at 25 ppt with yields of 0.99, 2.54 pg/ceU. A sharp decrease in all toxins profiles (CaTx, CmTx, HaTX and FjTx) was found at salinities of above 30 ppt.

Water quality of two semi-closed areas in the United Arab Emirates Coast along the Arabian Gulf:a comparative study

A comparative study was carried out between the two biggest creeks along the Arabian Gulf coast of the United Arab Emirates to evaluate impacts of sewage and industrial effluents on their hydrochemical characteristics. Surface and bottom water samples were collected from Abu Dhabi and Dubai creeks during the period from October 1994 to September 1995. The hydrochemical parameters studied were: temperature (21.10-34.00°C), salinity (37.37-47.09%), transparency (0.50-10.0 m), pH (7.97-8.83), dissolved oxygen (1.78-13.93 mg/l) and nutrients ammonia (ND- 13.12,ug-at N/1), nitrite (ND-6.66 ,ug-at N/1), nitrate (ND- 41.18 ,ug-at N/1), phosphate (ND- 13.06 ,ug-at P/1), silicate (0.68-32.50 ,ug-at Si/1), total phosphorus (0.26- 21.48 ,ug-at P/1), and total silicon (0.95- 40.32 ,ug-at Si/1). The present study indicates clearly that seawater of Abu-Dhabi Creek was warmer (28.l2°C) than Dubai (27.56°C) resulting in a higher rate of evaporation. Owing to more evaporation, salinity levels showed higher levels at Abu Dhabi (43.33%) compared to Dubai (39.03%) seawater. The study also revealed higher secchi disc readings at Abu Dhabi Creek (4.68 m) as compared to Dubai Creek (2.60 m) suggesting more transparency at Abu Dhabi Creek. Whereas, seawater of Dubai exhibited higher levels of pH (1.03 times), and dissolved oxygen (1.05 times) than Abu Dhabi seawater due to an increase in productivity. Meantime, seawater of Dubai showed higher tendency to accumulate ammonia (8.22 times), nitrite (10.93 times), nitrate (5.85 times), phosphate (10.64 times), silicate (1.60 times), total phosphorus (3.19 times), and total silicon (1.54 times) compared to Abu Dhabi seawater due to the enrichment of seawater at Dubai with domestic sewage waters which has distinctly elevated the levels of the nutrient salts particularly in inner-most parts of the creek leading to eutrophication signs. The changes occurred in the receiving creek water of Dubai as a result of waste-water disposal that have also reflected on the atomic ratios of nit: Effect of pollution rogen: phosphorus: silicon.

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.

Investigations on long distance transportation of fish. 4. A comparative study of the performance of expanded polystyrene slabs and multi-layer gunny (jute) fabric as insulants

A comparative study of the insulation efficiencies of expanded polystyrene slabs and multi-layer gunny fabric in long distance transportation of fresh iced fish was made. Used plywood boxes (second hand tea chests) were employed as containers and the experiments conducted between Kakinada and Calcutta. All the three insulants tried, namely, 25.4 mm thick expanded polystyrene slab, four and two layer gunny (jute) fabric, all sealed in 150 gauge polythene sheets, showed comparable insulation efficiencies, considering total bacterial counts, organoleptic qualities and TMA and TVN values of the transported fish as parameters.

Comparative study of three short-chain neurotoxins from the venom of Naja kaouthia (Yunnan, China)

Three short-chain neurotoxins named NT-I, NT-II, and NT-III were purified from the venom of Naja kaouthia, a snake distributed throughout the south of Yunnan province, China, by a series of chromatographic steps, including an FPLC Resource S column. Their molecular weights, determined by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) MS, were 6952.19 Da, 6854.92 Da, and 6828.80 Da, respectively. NT-I consisted of 62 amino acid residues, and the other two consisted of 61 amino acid residues, including 8 cysteines. After hydrolysis by endoproteinase Glu-C, their primary sequences were determined. A test of their activities demonstrated that they effectively inhibited muscle contractions induced by electric stimulation. Furthermore, the extent of inhibition caused by NT-II and NT-III was less than that of NT-I. The IC(50)s were 0.04 mug/ml, 0.20 mug/ml, and 0.23 mug/ml for NT-I, NT-II, and NT-III, respectively. Compared with NT-II and NT-III, the higher activity of NT-I may be a result of the amino acid residue substitution Ile36 to Arg36.

Comparative study of traditional and improved containers for transportation of fresh fish

Traditional bamboo basket and expanded polystyrene insulated plywood box (second hand tea-chest) were compared for transportation of iced marine, fresh water and brackish water fishes from Kakinada to Madras by rail. Quality of fish at dispatching and receiving centres was assessed by organoleptic, total volatile nitrogen (TVN) and total bacterial count (TBC) tests. Based on the results obtained and the prices fetched, the traditional bamboo basket apart from being cheaper was found to be as good as expanded polystyrene insulated plywood box for short distance transportation of iced fish involving less than 24 hours journey.

Comparative study of the nutrient content of fish and shell fish

The amino acid, mineral and proximate composition of mullet (Mugil oeur), mackerel (Rastrelliger kanagurta), crab (Scylla serrata) and prawn (Penaeus indicus) are reported. The data are used for comparing the nutritional quality of the fish and shell fish. Further, the amino acid composition is screened for their adequacy to meet the FAO/WHO recommended pattern of essential amino acids.

Length-weight relationship of two palaemonid prawns, Macrobrachium idella and M. scabriculum: a comparative study

Length-weight relationships of two commercially important freshwater prawns, Macrobrachium idella and M. scabriculum, have been worked out separately for the two sexes of the two species. 't' test showed that growth departs significantly from the isometric growth in the case of both sexes of M. idella and in the female of M. scabriculum. Therefore, the cubic formula W = CL^3 may be applied only in the case of males of M. scabriculum. In all the other cases W = CL^n may be followed.

A comparative study of various extenders of milkfish, Chanos chanos (Forsskal) sperm preservation

Four chemical extenders in 7 different concentrations (potassium chloride, sodium chloride, glucose, sodium citrate, Ringer s solution, cow serum and milkfish (Chanos chanos) serum) were compared in the preservation of milkfish sperm. Results showed milkfish serum to be the most suitable of the various extenders tested. This may be attributed to suitable osmotic potential and/or presence of proteins which may have directly or indirectly influenced sperm viability. The effects of milkfish serum on the motility and fertilizing capacity of sperm at different durations of storage however need to be investigated.

Quality of cured fish on the west coast: comparative study with the IS specifications

Moisture and salt content in cured fish products from various centres on the west coast of India are compared. The moisture contents varied in samples from different centres, whereas the salt content remained more or less uniform. The deviations from the Indian standard specification were highly significant in both cases. The high mean moisture values and low salt values with respect to accepted standards are indicative of the improper drying and poor salting.

A comparative study on the induced breeding and in vitro fertilization performance by various inducing agents in catfish, Heteropneustes fossilis (Bloch)

Brood catfish, Heteropneustes fossilis were collected from Tamirabarani river basin of Tamil Nadu, India and kept in cement tanks. Three inducing hormones viz, Ovaprim, Ovatide and WOVA-FH were injected at the rate of 0.5, 1.0, 1.5 and 2.0 ml/kg body weight in order to induce oocyte maturation and ovulation. After 10-13h of injection at a water temperature of 27±-0.5°C, stripping of eggs and in vitro fertilization was done. Ovaprim gave maximum (94.67%) hatching rate followed by Ovatide (90.33%) and WOVA-FH (77.33%).

Comparative study of seasonal changes in some blood parameters and genetic polymorphisms of serum antibodies and antigens on red blood cells surface in immature Caspian salmon

Seasonal sampling from 40 immature Caspian salmon were performed in summer, autumn, winter and spring. The maximum ranges of RBC counts, Hct, Hb, WBC count and clotting times were observed in spring, summer, spring, spring and winter, respectively. The minimum amounts of these factors were counted in summer, winter, winter, winter and winter, respectively. Blood Samples were taken from healthy smolt, immature and adult Caspian salmon in spawning time. Hematological determinations and biochemical serum analysis were performed in 101 fish in the three samples. The ranges of hematological values for sample mean were counted. Red blood cell counts were 866600 mm3 and 1259400 mm3 in smolt and adult respectively. Hematocrit was 48.39% in smolt and 44.29% in adult. Hemoglobin was 8.85 gr/dl in smolt and 10.91 gr/dl in adult. White blood cell count was 8781.58 mm3 in smolt and 5217.55 mm3 in adult and mean were differential of WBC, Lymphocyte 90.57%in smolt and73.22% in adult. Neutrophil was 5.12% in smolt and 16.92% in adult, Monocyte were 1.27% in smolt and 4.24% in adult, Clotting time was 282.34 Seconds in smolt and 291.47 seconds in adult MCV, MCH and MCHC also meagered in smolt and adult. Biochemical parameter in immature and mature Caspian salmon meagered .Glucose concentration was 2.97 mmol.l- in immature and 1.99 mmol.l- in mature .Cholesterol concentration was 4.26 mmol.l- in immature and 7.06 mmol.l- in mature. Triglyceride amount was 2.35 mmol.l- in immature and 2.47 mmol.l- in mature and Calcium was 2.47 in immature and 2.61 mmol.l- in mature. An in situ study was made on erythrocytic isoantigens and hetero-antigen and their corresponding iso-and hetero-antibodies of sera by means of hemoagglutination tests on the blood sample, of 450 immature and 50 mature Caspian salmon. The absence of erythrocyte iso-antigens and hetero-antigen and their corresponding iso-and hetero-antibodies were shown by the experimental. It could be indicated an intra-specific variation and differences in species for kelardasht hatchery.

A comparative study on effects of Immunoster and Immunowall as immunostimulants on some hematological, biochemical and growth indices of farmed great sturgeon juveniles (Huso huso)

Prebiotics are non-digestible food ingredients that profitably affect the host by selectively stimulating the growth and /or activation of one or a limited number of bacteria in the intestine that can enhance host health status. Immunoster (IS) and Immunowall (IW) are prebiotics and immunostimulants derived from the outer cell wall of brewers yeast, Saccharomyces cerevisiae. These substances contain MOS and �-glucans. After a four-week acclimatization period to rearing conditions and basal diet, 450 farmed great sturgeon juveniles weighing 95.58 ± 9.38 g were randomly distributed into 15 fiberglass tanks (2 × 2 × 0.53 m) in five treatments (Control, IS 1%, IW 1%, IS 3%, and IW 3%) in three replicates (completely randomized design) and kept at a density of 30 fish per tank for a period of 8 weeks at water temperature 20.55 ± 5.11ºC, dissolved oxygen 6.73 ± 0.35 mg L-1 and pH 7.92 ± 0.09. IS and IW were added at two levels of 1% and 3% to the basal diet in place of cellulose, except the control. At the beginning, in the middle and at the end of the trial, carcass analysis was done to determine the moisture, protein, fat, ash, and total carbohydrate. Also, blood samples were collected to measure hematological, biochemical and immune indices. At the end of the trial, final weight, final length, body weight increase (BWI), specific growth rate (SGR), average daily growth (ADG), protein efficiency ratio (PER), feed conversion ratio (FCR), and condition factor (CF) in fish fed on IS and IW in both levels 1% and 3% showed some differences. These differences were significant in IS 3% and IW 1% and 3% compared with the control (P<0.05). HSI showed no significant difference (P>0.05) and survival rate was 100% in all treatments. Crude protein of carcass in fish fed on IS and IW at 1% and 3% showed an increase in comparison with the control. There was significant difference between IS 3% and the control in crude protein of carcass (P<0.05). Fish fed on IS and IW at 1% and 3% showed various results in hematological and biochemical factors. It was observed significant difference in MCV between IW 1% and IS 3% compared with the control (P<0.05). Although there was an increase in values of hematocrit, hemoglobin (except IS 1%), WBC (except IW 3%), MCH, neutrophil, total protein, albumin (except IS 3%), K+, and lysozyme in fish fed on IS and IW compared with the control, it was no significant (P>0.05). The maximum count of WBC and the highest value of Ca2+ were seen in IW 1%. The maximum count of lymphocyte, the highest values of total protein, albumin and IgM were recorded in IW 3%. IS 1% had the maximum count of neutrophil and the highest concentration of lysozyme. Based on obtained results, it can be declared that IS and IW at two levels of 1% and 3% can enhance growth performance and feed efficiency and also improve some hematological, biochemical, and immune indices in farmed great sturgeon juveniles.