Acute effect of copper on Puntius javanicus survival and a current opinion for future biomarker development

Puntius javanicus experimental groups were exposed with to different concentration of copper (II) sulfate for 96 hours. Their mortality was recorded to determine LC50 value of copper concentration based on arithmetic, logarithmic and probit graphic analyses. The results obtained from these three mathematical analyses were 11.37±0.58, 11.01±0.73 and 10.68 mg/L, respectively. From the present study, we suggested that in the future, the range of 0 to 5.0 mg/L can be used to study the effect of copper concentration on fish activity at biochemical and physiological levels. Based on probit analysis, this maximum range is lower than LC10 value i.e. 6.11 mg/L. Therefore, it can be positively hypothesised that there would be no mortality occur except for several symptoms of adverse effects beyond of 5.0 mg/L treatment

Acute effect of copper on Puntius javanicus survival and a current opinion for future biomarker development

Puntius javanicus experimental groups were exposed with to different concentration of copper (II) sulfate for 96 hours. Their mortality was recorded to determine LC50 value of copper concentration based on arithmetic, logarithmic and probit graphic analyses. The results obtained from these three mathematical analyses were 11.37±0.58, 11.01±0.73 and 10.68 mg/L, respectively. From the present study, we suggested that in the future, the range of 0 to 5.0 mg/L can be used to study the effect of copper concentration on fish activity at biochemical and physiological levels. Based on probit analysis, this maximum range is lower than LC10 value i.e. 6.11 mg/L. Therefore, it can be positively hypothesised that there would be no mortality occur except for several symptoms of adverse effects beyond of 5.0 mg/L treatment

Evaluation of acetylcholinesterase source from fish, Tor tambroides for detection of carbamate

Acetylcholinesterase (AChE) from the brain tissue of local freshwater fish, was isolated through affinity purification. Acetylthiocholine iodide (ATCi) was preferable synthetic substrate to purified AChE with highest maximal velocity (V ) and lowest biomolecular constant (K ) at 113.60 Umg and 0.0689 mM, respectively, with highest catalytic efficiency ratio (V /K ) of 1648.77. The optimum pH was 7.5 with sodium phosphate buffer as medium, while optimal temperature was in the range of 25 to 35 °C. Bendiocarp, carbofuran, carbaryl, methomyl and propoxur significantly lowered the AChE activity greater than 50%, and the IC value was estimated at inhibitor concentration of 0.0758, 0.0643, 0.0555, 0.0817 and 0.0538 ppm, respectively

Purification and anticholinesterase sensitivity of cholinesterase extracted from liver tissue of Puntius javanicus

The purification of a soluble cholinesterase (ChE) from Puntius javanicus liver using affinity chromatography was studied. Affinity matrix was synthesised through the cooling system of ligands procainamide to epoxy-activated Sephacryl 6B and purification process was performed using calibrated flow rate at 0.2 mL/min. Non-denaturing electrophoresis condition was employed and the single band native form of ChE was detected at 66.267 kDa after being stained with commasie brilliant blue. ChE detection was performed using gel filtration; ZORBAX column attached to the HPLC with the flow rate of 1 mL/min. Only a single peak was detected at the retention time of 3.720. From the assay evaluation, the final purified ChE procedure displayed the highest sensitivity of detecting the anticholinesterase namely mercury, copper, malaoxon and carbofuran compared to the impure ChE and the results were further discussed in detail to the potential application of ChE from P. javanicus as a biomarker for those toxicants International Journal of Agriculture and Biolog

Purification and anticholinesterase sensitivity of cholinesterase extracted from liver tissue of Puntius javanicus

The purification of a soluble cholinesterase (ChE) from Puntius javanicus liver using affinity chromatography was studied. Affinity matrix was synthesised through the cooling system of ligands procainamide to epoxy-activated Sephacryl 6B and purification process was performed using calibrated flow rate at 0.2 mL/min. Non-denaturing electrophoresis condition was employed and the single band native form of ChE was detected at 66.267 kDa after being stained with commasie brilliant blue. ChE detection was performed using gel filtration; ZORBAX column attached to the HPLC with the flow rate of 1 mL/min. Only a single peak was detected at the retention time of 3.720. From the assay evaluation, the final purified ChE procedure displayed the highest sensitivity of detecting the anticholinesterase namely mercury, copper, malaoxon and carbofuran compared to the impure ChE and the results were further discussed in detail to the potential application of ChE from P. javanicus as a biomarker for those toxicants

Hexavalent molybdenum reduction to molybdenum blue by S. marcescens Strain Dr. Y6

A molybdate-reducing bacterium has been locally isolated. The bacterium reduces molybdate or Mo(6+) to molybdenum blue (molybdate oxidation states of between 5+ and 6+). Different carbon sources such as acetate, formate, glycerol, citric acid, lactose, fructose, glucose, mannitol, tartarate, maltose, sucrose, and starch were used at an initial concentration of 0.2% (w/v) in low phosphate media to study their effect on the molybdate reduction efficiency of bacterium. All of the carbon sources supported cellular growth, but only sucrose, maltose, glucose, and glycerol (in decreasing order) supported molybdate reduction after 24 h of incubation. Optimum concentration of sucrose for molybdate reduction is 1.0% (w/v) after 24 h of static incubation. Ammonium sulfate, ammonium chloride, valine, OH-proline, glutamic acid, and alanine (in the order of decreasing efficiency) supported molybdate reduction with ammonium sulfate giving the highest amount of molybdenum blue after 24 h of incubation at 0.3% (w/v). The optimum molybdate concentration that supports molybdate reduction is between 15 and 25 mM. Molybdate reduction is optimum at 35 degrees C. Phosphate at concentrations higher than 5 mM strongly inhibits molybdate reduction. The molybdenum blue produced from cellular reduction exhibits a unique absorption spectrum with a maximum peak at 865 nm and a shoulder at 700 nm. The isolate was tentatively identified as Serratia marcescens Strain Dr.Y6 based on carbon utilization profiles using Biolog GN plates and partial 16s rDNA molecular phylogeny

Acute effect of copper on Puntius javanicus survival and a current opinion for future biomarker development

Puntius javanicus experimental groups were exposed with to different concentration of copper (II) sulfate for 96 hours. Their mortality was recorded to determine LC50 value of copper concentration based on arithmetic, logarithmic and probit graphic analyses. The results obtained from these three mathematical analyses were 11.37±0.58, 11.01±0.73 and 10.68 mg/L, respectively. From the present study, we suggested that in the future, the range of 0 to 5.0 mg/L can be used to study the effect of copper concentration on fish activity at biochemical and physiological levels. Based on probit analysis, this maximum range is lower than LC10 value i.e. 6.11 mg/L. Therefore, it can be positively hypothesised that there would be no mortality occur except for several symptoms of adverse effects beyond of 5.0 mg/L treatment

Comparison of Microtox and Xenoassay Light as a Near Real Time River Monitoring Assay for Heavy Metals

Luminescence-based assays for toxicants such as Microtox, ToxAlert, and Biotox have been used extensively worldwide. However, the use of these assays in near real time conditions is limited due to nonoptimal assay temperature for the tropical climate. An isolate that exhibits a high luminescence activity in a broad range of temperatures was successfully isolated from the mackerel, Rastrelliger kanagurta. This isolate was tentatively identified as Photobacterium sp. strain MIE, based on partial 16S rDNA molecular phylogeny. Optimum conditions that support high bioluminescence activity occurred between 24 and 30°C, with pH 5.5 to 7.5, 10 to 20 g/L of sodium chloride, 30 to 50 g/L of tryptone, and 4 g/L of glycerol as the carbon source. Assessment of near real time capability of this bacterial system, Xenoassay light to monitor heavy metals from a contaminated river running through the Juru River Basin shows near real time capability with assaying time of less than 30 minutes per samples. Samples returned to the lab were tested with a standard Microtox assay using Vibrio fishceri. Similar results were obtained to Xenoassay light that show temporal variation of copper concentration. Thus, this strain is suitable for near real time river monitoring of toxicants especially in the tropics

Kinetics of molybdenum reduction to molybdenum blue by Bacillus sp. strain A.rzi

Molybdenum is very toxic to agricultural animals. Mo-reducing bacterium can be used to immobilize soluble molybdenum to insoluble forms, reducing its toxicity in the process. In this work the isolation of a novel molybdate-reducing Gram positive bacterium tentatively identified as Bacillus sp. strain A.rzi from a metal-contaminated soil is reported. The cellular reduction of molybdate to molybdenum blue occurred optimally at 4 mM phosphate, using 1% (w/v) glucose, 50 mM molybdate, between 28 and 30°C and at pH 7.3. The spectrum of the Mo-blue product showed a maximum peak at 865 nm and a shoulder at 700 nm. Inhibitors of bacterial electron transport system (ETS) such as rotenone, sodium azide, antimycin A, and potassium cyanide could not inhibit the molybdenum-reducing activity. At 0.1 mM, mercury, copper, cadmium, arsenic, lead, chromium, cobalt, and zinc showed strong inhibition on molybdate reduction by crude enzyme. The best model that fitted the experimental data well was Luong followed by Haldane and Monod. The calculated value for Luong's constants pmax, Ks, Sm, and n was 5.88 mole Mo-blue hr-1, 70.36 mM, 108.22 mM, and 0.74, respectively. The characteristics of this bacterium make it an ideal tool for bioremediation of molybdenum pollution