Cloning and characterization of a blood coagulation factor IX-binding protein from the venom of Trimeresurus stejnegeri

A blood coagulation factor IX-binding protein (TSV-FIX-BP) was isolated from the snake venom of Trimeresurus stejnegeri. On SDS-polyacrylamide gel electrophoresis, TSV-FIX-BP showed a single band with an apparent molecular weight of 23,000 under non-reducing conditions. and two distinct bands with apparent molecular weights of 14,800 and 14,000 under reducing conditions. cDNA clones containing the coding sequences of TSV-FIX-BP were isolated and sequenced to determine the structure of the precusors of TSV-FIX-BP subunits. The deduced amino acid sequences of two subunits of TSV-FIX-BP were confirmed by N-terminal protein sequencing and trypsin-digested peptide mass fingerprinting. TSV-FIX-BP was a nonenzymatic C-type lectin-like anti-coagulant. The anti-coagulant activity of TSV-FIX-BP was mainly caused by its dose dependent interaction with blood coagulation factor IX but not with blood coagulation factor X. (C) 2003 Elsevier Science Ltd. All rights reserved.

Molecular cloning and characterization of a platelet glycoprotein Ib-binding protein from the venom of Trimeresurus stejnegeri

A platelet glycoprotein Ib-binding protein, termed TSV-GPIb-BP, was isolated from the venom of Trimeresurus stejnegeri. On SDS-polyacrylamide gel electrophoresis, TSV-GPIb-BP showed a single band with an apparent molecular weight of 28,000 and two distinct bands with apparent molecular weights of 16,000 and 15,000 under non-reducing and reducing conditions, respectively. cDNA clones containing the coding sequences for both TSV-GPIb-BP subunits were isolated and sequenced. The deduced amino acid sequences of TSV-GPIb-BP subunits were confirmed by N-terminal protein sequencing and trypsin-digested peptide mass fingerprinting. Interestingly, the a subunit of TSV-GPIb-BP is identical to that of alboaggregin-B, and the sequence identity of their beta subunits is 94.3%. TSV-GPIb-BP inhibited ristocetin-induced human platelet agglutination in platelet-rich plasma under lower dosages (<5 mug/ml). On the other hand, it directly aggregated washed human platelets in the absence of additional Ca2+ or any other cofactors under higher dosages (>5 mug/ml). This platelet aggregation activity was dose-dependently inhibited by specific GPIbalpha antibodies, but not by those antibodies against platelet GPIa, GPIIa, GPIIb and GPIIIa. (C) 2003 Elsevier Science Ltd. All rights reserved.

Development of low cost feed using local ingredients for culture of freshwater prawn, Macrobrachium rosenbergii in ponds

Two experiments were conducted to formulate and evaluate test diets using locally available ingredients to find out suitable diets for mono and polyculture of Macrobrachium rosenbergii in ponds. The first experiment was conducted from 1 July 2003 to 29 September 2003 in 12 experimental ponds each measuring 30 square meters behind the Fisheries Faculty Building, BAU campus, Mymensingh. Three experimental diets containing 30% protein were formulated using fish meal, meat and bone meal, mustard oilcake, sesame meal and rice bran and assigned to treatments 1, 2 and 3, respectively. A special shrimp feed (Starter-II) from Saudi-Bangla Fish Feed Ltd. was assigned to treatment 4 (Control). Each treatment had three replications. Juveniles of M. rosenbergii (2.90±0.81g) were stocked at the rate of 4/square meter. Prawns were fed three times daily at the rate of 15% of their body weight at the beginning, which was gradually reduced to 10% and 5% for the last two months. There was no significant (P>0.05) difference between the weight gains of prawns fed diets 1 and 4 (control), but they were significantly (P<0.05) higher than those of diets 2 and 3. The FCR values of diets ranged between 2.61 to 3.36 with diets 1 and 4 showing significantly (P<0.05) lower FCR values. The survival rate of prawns ranged between 68 to 78% with prawns fed diets 1 and 4 showing significantly higher survival rate. The production of prawn ranged from 921 to 1,428 kg/ha/90 days and diet 1 gave the highest production. Treatment 1 gave the highest net profit ofTk. 161,980/ha/90 days. The second experiment was conducted from December, 2003 to April, 2004 to see the growth of over wintered M. rosenbergii juveniles in polyculture with indigenous major carps (catla and rohu) in ponds using formulated diets. Three isoenergetic experimental diets formulated using fish meal, meat and bone meal, mustard oil cake, rice bran, wheat bran and molasses, and a shrimp feed 'Golda special feed' from Saudi-Bangla Fish Feed Ltd. were assigned to treatments 1, 2, 3 and 4 (control), respectively, each treatment had two replications. Eight experimental ponds each measuring 80 square meters in the Field Laboratory Complex of the Faculty of Fisheries, BAU campus, Mymensingh were used. The mean initial weights of M. rosenbergii, catla and rohu were 1.60±0.01, 30.0±0.09 and 25.0±0.08 g, respectively. A total of 160 fish and prawn (20,000/ha) were stocked in each pond at the ratio of 2:1:1 (prawn: catla: rohu). Fish were fed at the rate of 3-5% of their body weight. Prawns fed diet 1, 2 and 4 showed higher weight gains compared to diet 3. The weight gain of catla was significantly higher in T1 while in case of rohu was higher in T1 and T4, respectively. There were no significant differences (P>0.05) in the survival rate of fish as obtained from different treatments. The overall total fish production ranged from 2196 to 2679 kg/ha/5 months. The highest production and the highest profit (Tk. 56,531/ha/5 months) was obtained from T1 and the lowest (Tk. 24,932/ha/5 months) from T4.

Effect of stocking density on the production of common carp (Cyprinus carpio Lin.) in cages at Kaptai lake, Bangladesh

Growth, survival and food conversion ratio (FCR) of Cyprinus carpio in cages at different stocking density were studied in Kaptai lake. Fingerlings of common carp (12.47 ± 4.29 g) were stocked at 25/m2 , 50/m2 and 75/m2 each with replication in six floating net cages each of Sm x Sm x 4m and reared for 240 days. The growth rate was inversely related to the stocking density with the mean weights of 325.5 ± 11.74 g, 268.99± 8.44 g and 167.0 ± 11.66 g at low, medium and high densities, respectively. At harvest, standing crop biomass averaged 7 .82, 12.83 and 11.58 kg with the survival of 98.9%, 97.6% and 94.4% and food conversion ratios of 4.51, 3.82 and 4.21 for the above three densities, respectively. Weight gain and production at density 25 fish/m 2 were significantly different (p< 0.01) from other two densities. Water quality was not affected in the cages having different stocking densities.

Structured precision modelling with Cholesky basis superposition for speech recognition

Structured precision modelling is an important approach to improve the intra-frame correlation modelling of the standard HMM, where Gaussian mixture model with diagonal covariance are used. Previous work has all been focused on direct structured representation of the precision matrices. In this paper, a new framework is proposed, where the structure of the Cholesky square root of the precision matrix is investigated, referred to as Cholesky Basis Superposition (CBS). Each Cholesky matrix associated with a particular Gaussian distribution is represented as a linear combination of a set of Gaussian independent basis upper-triangular matrices. Efficient optimization methods are derived for both combination weights and basis matrices. Experiments on a Chinese dictation task showed that the proposed approach can significantly outperformed the direct structured precision modelling with similar number of parameters as well as full covariance modelling. © 2011 IEEE.

Activation mechanism of pre-phenoloxidase in lobster and shrimp

Phenoloxidase in shrimp and lobster is shown to exist in latent form which could be activated by trypsin and by an endogenous enzyme with tryptic activity. On Sephadex G-100 gel, three isoenzymes, differing in molecular weights, were isolated from naturally activated lobster shell extracts. Mechanism of activation of ptephenoloxidase involving limited proteolysis by the activating enzyme to form isoenzymes has been proposed.

Purification and cloning of a novel C-type lectin-like protein with platelet aggregation activity from Trimeresurus mucrosquamatus venom

TMVA, a novel C-type lectin-like protein that induces platelet aggregation in a dose-dependent manner, was purified from the venom of Trimeresurus mucrosquamatus. It consists of two subunits, alpha (15,536 Da) and beta (14,873 Da). The mature amino acid sequences of the a (135 amino acids) and beta subunits (123 amino acids) were deduced from cloned cDNAs. Both of the sequences show great similarity to C-type lectin-like venom proteins, including a carbohydrate recognition domain. The cysteine residues of TMVA are conserved at positions corresponding to those of flavocetin-A and convulxin, including the additional Cys135 in the alpha subunit and Cys3 in the beta subunit. SDS-PAGE, mass spectrometry analysis and amino acid sequence showed that native TMVA exists as two convertible multimers Of (alphabeta)(2) and (alphabeta)(4) with molecular weights of 63,680 and 128,518 Da, respectively. The (alphabeta)(2) complex is stabilized by an interchain disulfide bridge between the two alphabeta-heterodimers, whereas the stabilization of the (alphabeta)(4) complex seems to involve non-covalent interactions between the (alphabeta)(2) complexes. (C) 2002 Elsevier Science Ltd. All rights reserved.

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.

Molecular characterization of a weak fibrinogen-clotting enzyme from Trimeresurus jerdonii venom

A fibrinogen-clotting enzyme designed as jerdonobin-II was isolated from the venom of Trimeresurus jerdonii. It differed in molecular weight and N-terminal sequence with the previously isolated jerdonobin, a thrombin-like enzyme from the same venom. The enzyme consists of a single polypeptide chain with molecular weights of 30,000 and 32,000 under non-reducing and reducing conditions, respectively. Jerdonobin-II showed weak fibrinogen clotting activity and its activity unit on fibrinogen was calculated to be less than one unit using human thrombin as standard. The precursor protein sequence of jerodonobin-II was deduced from cloned cDNA sequence. The sequence shows high similarity (identity = 89%) to TSV-PA, a specific plasminogen activator from venom of T stejnegeri. Despite of the sequence similarity, jerdonobin-II was found devoid of plasminogen activating effect. Sequence alignment analysis suggested that the replacement of Lys(239) in TSV-PA to Gln(239) in jerdonobin-II might play an important role on their plasminogen activating activity difference. (C) 2005 Elsevier Ltd. All rights reserved.

Standardisation of specification for different trolling lures

Fishing experiments were conducted with buffalo horn, wooden and galvanised iron jigs of different lengths and weights rigged in troll lines. It was observed that jigs with 12 cm length and 60 g weight were superior to others in luring seer fish. Fish head jigs of 60 g weight showed a higher efficiency index than those of 50 and 70 g tried in these studies.

On further improvement of the techinque of evaluating summer institutes

Data from an earlier study are reanalyzed to improve upon the evaluation measures of summer institutes. The Summer Institute Efficiency Index is improved by using weighted geometric mean using different ranks as weights for the several dimensions. The coverage utility index is improved by taking the ratings given by all the participants.

Growth and survival of Penaeus monodon postlarvae fed shrimp head meal and fish meal as primary animal source of protein

Although shrimp head meal alone does not provide for good growth and survival, fish meal can provide high survival rate. The addition of shrimp head improves this diet. It is suggested that cholesterol present in shrimp could have caused this difference. Composition of the test diets is tabulated, as are proximate chemical analysis of the diets, and the mean initial weights, final weights, weight gains, survival rate, feed consumed, protein consumed, of Penaeus monodon postlarvae, feed conversion and protein efficiency ratio.

Nutritional evaluation of texturised meat from Nemipterus japonicus

Evaluation of the nutritional quality of texturised fish meat from threadfin bream (Nemipterus japonicus) is reported. Proximate composition and amino acid analysis showed that its protein content is very high with all the essential amino acids present in higher levels than those prescribed for man in the 1973 FAO/WHO amino acid scoring pattern. Lysine amounted to 11.0 g per 16 g N of which 75 percent is biologically available. Studies on PER and NPU also corroborate the excellent nutritional quality of the texturised meat.

Effect of delayed icing on the quality of tiger shrimp (Penaeus monodon Fab.)

Effect of delayed icing on the quality of Penaeus monodon iced after three hours of harvest was studied in plastic and bamboo baskets. After harvest of three hours at ambient temperature (28°-32°C), ice was added to the shrimp at a ratio of 1:1 (shrimp:ice) and stored for 21 hours in both the baskets. Quality evaluation was carried out through visual assessment, biochemical analysis and microbial analysis for 24 hours. The organoleptic evaluation and scoring was done from the time of harvest treated as 0 hour and the average score was 10. At 9th hour after iced condition quality of shrimp was found reduced to the next stage (acceptable) with a score ranged from 8.4-6.5 in both baskets. This acceptable stage was observed throughout the experiment for bamboo basket whereas in the plastic basket the quality was reduced to a small extent with a score of 6.4 (moderately acceptable). Till the end point of the experiment the quality of shrimp was acceptable in respect to biochemical analysis. The microbial load was found log sub(10) 3.99±0.12 cfu/g to log sub(10) 4.33±0.21 cfu/g and log sub(10) 4.01 ±0.12 cfu/g to log sub(10) 4.83±0.19 cfu/g in the bamboo and plastic basket respectively. The importers or buyers suggests for immediate icing to maintain good quality but results of the present experiment suggest that the quality does not vary drastically for first three hours.

Culture potentials of climbing perch, Anabas testudineus (Bloch) under different stocking densities at semi-intensive management

Anabas testudineus was cultured at different stocking density for the period of five months from May to September. Three stocking densities such as 50,000 (Treatment-1, T1), 56,250 (Treatment-2, T2) and 62,250/ha (Treatment-3, T3) were tested with three replications. After five months rearing, the mean weights of koi were 46.74±2.59, 40.44±2.98 and 37.27±3.01 in T-1, 2 and 3, respectively. The calculated production of native koi in T1 T2 and T3 were 1,916±314, 1,774.31 ±260 and 1,431 ±297 kg/ha, respectively which were significantly different (p<0.05) from each other.