A new alternative local source of acetylcholinesterase (AChE) has been found to be sensitive towards several insecticides. AChE was purified from the brain tissue of Puntius javanicususing affinity chromatography (procainamide hydrochloride coupled with sephacryl 6B) with the total recovery of 40% at 8.48 purification fold. All carbamate (CB) insecticides tested at the concentration of 1 mg/L were capable of lowering AChE activity to less than 30%; Bendiocarp (18.80%), carbaryl (5.96%,), carbofuran (6.12%), methomyl (13.91%) and propoxur (15.851%). The P. javanicus AChE was found to be unaffected by several activated organophosphates (OP) such as acephate and dimethoate, while trichlorfon slightly inhibited the enzyme activity (p<0.05). Chlorpyrifos, diazinon, malathion and parathion lowered AChE activity to 43.02, 40.97, 37.11 and 46.72%, respectively. Pesticides that inhibit AChE activities more than 50% were further tested in different concentrations to determine the half maximal inhibitory concentration (IC50). Carbofuran and carbaryl showed the lowest IC50 value at 0.035 and 0.031 mg/L, respectively, as both showed no significant differences (p<0.05), followed by bendiocarp, propoxur, methomyl, malathion, diazinon, parathion and chlorpyrifos at 0.045, 0.076, 0.090, 0.063, 0.103, 0.151 and 0.202 mg/L, respectively. Based on these results, the sensitivity of AChE from brain P. javanicus brain tissue shows promise as an alternative biosensor for the detection of insecticides contamination.

The environmental toxicants such as copper are known to affect vital organ especially liver. This study examined the effects of copper sulfate (CuSO 4 ) on the liver morphological structure of P. javanicus. The untreated control, 0.1 and 0.3 mg/L CuSO 4 treated groups displayed normal polygonal structure of the hepatocyte. However, at the concentrations of 0.5, 1.0 and 5.0 mg/L CuSO 4 , the hepatostructure was significantly affected, as shown by the increasing number of dilation and congestion of sinusoids, vacuolation, macrophage activities and peliosis. The damage level and HSI value were increased while the number of hepatic nuclei per mm 2 was decreased with the increasing of copper concentration. In conclusion, this study shows that the degree of liver damage in P.javanicus is dependent to the dose exposure. � All Rights Reserved.

One of the most economical approaches for removal of toxic compounds is bioremediation. In the long term, bioremediation is economic and feasible compared to other methods, such as physical or chemical methods. A bacterium that can efficiently reduce molybdenum blue was isolated from polluted soil. Biochemical analysis revealed the identity of the bacterium as Enterobacter sp. strain Saw-1. The growth parameters for optimal reduction of molybdenum to Mo-blue or molybdenum blue, a less toxic product, were determined around pH 6.0 to 6.5 and in the range of 30 to 37 �C, respectively. Glucose was selected as preferred carbon source, followed by sucrose, maltose, l-rhamnose, cellobiose, melibiose, raffinose, d-mannose, lactose, glycerol, d-adonitol, d-mannitol, l-arabinose and mucate. Phosphate and molybdate were critically required at 5.0 mM and 10 mM, respectively. The scanning absorption spectrum acquired to detect the development of complex Mo-blue showed similarity to previously isolated Mo-reducing bacteria. In addition, the spectrum closely resembled the molybdenum blue from the phosphate determination method. Heavy metals, including mercury, copper (II) and silver (I), inhibited reduction. Moreover, the bacterium also showed capability of exploiting the pesticide coumaphos as an alternative carbon source for growth. As the bacterium proved its ability to detoxify organic and inorganic xenobiotics, the usefulness of this microorganism for bioremediation is highlighted. � 2017 Published by ITB Journal Publisher.

Crude extract of ChE from the liver of Puntius javanicus was purified using procainamide-sepharyl 6B. S-Butyrylthiocholine iodide (BTC) was selected as the specific synthetic substrate for this assay with the highest maximal velocity and lowest biomolecular constant at 53.49 μmole/min/mg and 0.23 mM, respectively, with catalytic efficiency ratio of 0.23. The optimum parameter was obtained at pH 7.5 and optimal temperature in the range of 25 to 30°C. The effect of different storage condition was assessed where ChE activity was significantly decreased after 9 days of storage at room temperature. However, ChE activity showed no significant difference when stored at 4.0, 0, and -25°C for 15 days. Screening of heavy metals shows that chromium, copper, and mercury strongly inhibited P. javanicus ChE by lowering the activity below 50%, while several pairwise combination of metal ions exhibited synergistic inhibiting effects on the enzyme which is greater than single exposure especially chromium, copper, and mercury. The results showed that P. javanicus ChE has the potential to be used as a biosensor for the detection of metal ions.

Due to the latest industrial development, many dangerous chemicals have been released directly or indirectly which resulted in the polluted water bodies. Water rehabilitation is an alternative way to restore the quality of water, followed by the environmental management to control the waste discharge to ensure the balance of the degradation rates or detoxifying by environmental factors. However, this process consumed a lot of time and cost. Besides, most of the metal ions, especially copper which is capable to bioaccumulate in aquatic organism and at the elevated level may cause physiological and biochemical alteration which leads to mortality. Environmental monitoring is the initial step presupposed evaluating the potential toxicity of effluent gushing at its purpose to discharge, avoiding the determining effects of contaminant in water bodies. Due to the high sensitivity of the aquatic life towards dissolving toxicant, the fish has been utilized as the biological measurement (Biomarker) to indicate the existence of toxicant exposure and/or the impact towards the evaluation of molecular, cellular to physiological level. Thus, this paper gives an overview of the manipulation of fish as a biomarker of heavy metals through behavior response, hepatocyte alteration, enzymatic reaction and proteomic studies which have proven to be very useful in the environmental pollution monitoring. � All Rights Reserved.

The aim of this study is to determine the inhibitory effect of copper towards butyrylcholinesterase (BChE) activity. Using the Lineweaver-Burk plot, Puntius javanicus BChE activity was found to be noncompetitively inhibited by copper. The maximal velocities of untreated (control) BChE, 0.5 and 10 mg/L copper-treated BChE are 53.70, 31.81 and 14.30 Umg-1, respectively, while the biomolecular constant (Km) values of both tests shows no significant difference (p>0.05). The in vitro IC50 of copper ion to the BChE was found to be 0.0948 (0.0658 to 0.1691) mg/L. In vivo tests showed that in the presence of 0.1 mg/L copper, the BChE activity was slightly higher compared to the untreated control. Copper sulfate at 0.3 mg/L concentration showed no significant inhibition compared to control. However, the activity decreased with increasing copper concentrations of 0.5, 1.0 and 5.0 mg/L, with the remaining activity at 87.60, 84.60 and 73.00 %, respectively. This study suggests that BChE isolated from P. javanicus liver tissue is a potentially new source of biomarker for copper contamination. � by PSP.

Bioremediation of pollutants including heavy metals and xenobiotics is an economic and environmentally friendly process. A novel molyb-denum-reducing bacterium with the ability to utilize the pesticide glyphosate as a carbon source is reported. The characterization works were carried out utilizing bacterial resting cells in a microplate format. The bacterium reduces molybdate to Mo-blue optimally between pH 6.3 and 6.8 and at 34oC. Glucose was the best elec-tron donor for supporting molybdate reduction followed by lactose, maltose, melibiose, raffinose, d-mannitol, d-xylose, l-rhamnose, l-arabinose, dulcitol, myo-inositol and glycerol in descending order. Other requirements include a phosphate concentration at 5.0 mM and a molybdate concentration between 20 and 30 mM. The molybdenum blue exhibited an absorption spec-trum resembling a reduced phospho-molybdate. Molybdenum reduction was inhibited by mercury, silver, cadmium and copper at 2 ppm by 45.5, 26.0, 18.5 and 16.3%, respectively. Biochemical analysis identified the bacterium as Klebsiella oxytoca strain Saw-5. To conclude, the capacity of this bacterium to reduce molybdenum into a less toxic form and to grow on glyphosate is novel and makes the bacterium an important instrument for bioremediation of these pollutants. � 2016, Agriculture Faculty Brawijaya University. All rights reserved.

Molybdenum is an emerging pollutant worldwide. The objective of this study is to isolate molybdenum-reducing bacterium with the ability to grow on phenolic compounds (phenol and catechol). The screening process was carried out on a microplate. The bacterium reduced molybdenum in the form of sodium molybdate to molybdenum blue (Mo-blue). The bacterium required a narrow pH range for optimal reduction of molybdenum, i.e. Between pH 6.3 and 6.8, with temperature between 34 and 37 o C. Molybdate reduction to Mo-blue was best supported by glucose as the carbon source. However, both phenol and catechol could not support molybdate reduction. Other requirements for molybdate reduction included sodium molybdate concentrations between 15 and 30 mM, and phosphate concentration of 5.0 mM. The bacterium exhibited a Mo-blue absorption spectrum with a shoulder at 700 nm and a maximum peak near the infrared region at 865 nm. The Mo-reducing bacterium was partially identified as Enterobacter sp. Strain Saw-2. The capability of this bacterium to grow on toxic phenolic compounds and to detoxify molybdenum made it a significant agent for bioremediation.

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. � 2015 Friends Science Publishers.