OBJECTIVETo investigate the diversity of acetylcholinesterase (AChE) activity in variety classes and strains of Culex pipiens pallens and provide a basis for the insecticide-resistance detection of mosquito by biochemical method.

METHODSAChE insensitivity of single mosquito was determined, using acetythiocholine iodide (ATch) as the substrate, 5,5'-dithio-bis (2-nitrobenzoic acid) (DTNB) as the developer, and propoxur as the inhibitor.

RESULTThere were significant differences in AChE activity among the four types of IV instar larvae and 3-day-old adult female of sensitive strain mosquito (P<0.01). The AChE activity of the 3-day-old adult female was higher than that of IV instar larvae of the four types of sensitive strain mosquito (P<0.01). The AChE activity of anti-DDVP (Rd) and anti-propoxur (Rp) strains of Culex pipiens pallens was significantly higher than that of sensitive (S) strain (P<0.01), while the AChE activity of anti-cypermethrin (Rc) strain of Culex pipiens pallens was similar to that of S strain (P>0.05). The individual frequency of insensitive AChE of Rd and Rp strains of Culex pipiens pallens was significantly higher than that of sensitive (S) strain (P<0.01), while the individual frequency of insensitive AChE of Rc strain of Culex pipiens pallens was similar to that of S strain(P>0.05).

CONCLUSIONThe AChE activity determination can be used to examine the insecticide-resistance of mosquito.

Acetylcholinesterase ; metabolism ; Animals ; Culex ; classification ; enzymology ; Dichlorvos ; pharmacology ; Female ; Insecticide Resistance ; Propoxur ; pharmacology ; Pyrethrins ; pharmacology ; Species Specificity

A modified electrometric method was described and validated for measurement of plasma and erythrocyte cholinesterase activities in 6~18 months old goats. The enzymatic reaction mixture contained 3 ml distilled water, 3 ml barbital-phosphate buffer (pH 8.1), 0.2 ml plasma or erythrocytes and 0.1 ml acetylthiocholine iodide (7.5%) as a substrate. The mixture was incubated at 37 degrees C for 40 minutes. The pH of the reaction mixture was determined by a pH meter before and after the incubation. The initial pH was measured before the substrate addition. The enzyme activity was expressed as deltapH/40 min. The coefficients of variation of the described method in measuring plasma and erythrocyte cholinesterase activities were 4 and 2%, respectively. Preliminary reference values (n = 14) of the mean cholinesterase activity (deltapH/40 min) and 95% confidence interval in the plasma were 0.194 and 0.184~ 0.204, respectively, and those of the erythrocytes were 0.416 and 0.396~0.436, respectively. The pseudocholinesterase activity of the plasma cholinesterase was 63.5% as determined by quinidine sulfate inhibition. The organophosphorus insecticides dichlorvos and diazinon at 0.5~4 micrometer and the carbamate insecticide carbaryl at 5~20 micrometer in the reaction mixture significantly inhibited plasma (13.7~85.5%) and erythrocyte (16.4~71.9%) cholinesterases in vitro in a concentration-dependent manner. The results suggest that the described electrometric method is simple, precise and efficient in measuring blood cholinesterase activity in goats.
Acid-Base Equilibrium/physiology ; Animals ; Carbaryl/pharmacology ; Cholinesterase Inhibitors/pharmacology ; Cholinesterases/*blood/drug effects ; Diazinon/pharmacology ; Dichlorvos/pharmacology ; Enzyme Activation/drug effects/physiology ; Erythrocytes/metabolism ; Goats/*blood ; Plasma/metabolism

OBJECTIVETo study the protective effect of memantine on the regulation of N-methyl-D-aspartate (NMDA) receptor in dichlorvos-poisoned rat brain, and to understand the mechanism of its role in organophosphate poisoning.

METHODSSD rats were administrated dichlorvos (25 mg/kg, ip) then three groups were treated with memantine at doses of 5, 15 and 45 mg/kg respectively. The activity of acetylcholinesterase (AChE) and binding capacity of NMDA receptor with [(3)H]MK-801 were determined 16 h after dichlorvos injection.

RESULTSThe time of onset of toxic symptoms in 15, 45 mg memantine treated groups [(18.40 +/- 1.14) and (21.40 +/- 1.52) min respectively] was higher than that in dichlorvos alone group [(16.75 +/- 1.62) min]; the intensity of muscle fasciculation (1.60 +/- 1.14 and 0.80 +/- 0.84, respectively) was less than that in control group (2.85 +/- 0.37); the total score of poisoning symptoms (8.80 +/- 1.79 and 9.00 +/- 2.24 respectively) was also less than that in dichlorvos group (14.60 +/- 1.70). The AChE activities both in blood and brain of memantine treated groups were not significantly different from those in dichlorvos alone group. The affinity (Kd value) and density (Bmax value) of brain NMDA receptor in dichlorvos exposed rats [(75.55 +/- 7.87) nmol/L, (0.46 +/- 0.06) pmol/mg pro respectively] were higher and lower respectively than those in control group [(37.37 +/- 4.17) nmol/L, (0.62 +/- 0.04) pmol/mg pro respectively]. Lower level of memantine (5 and 25 mg/kg) could antagonize the dichlorvos-evoked down-regulation of [(3)H]MK-801 binding to NMDA receptor in rat brain [Bmax value: (0.55 +/- 0.07) and (0.64 +/- 0.07) pmol/mg pro; Kd value: (38.68 +/- 4.54) and (32.58 +/- 3.90) nmol/L respectively] while higher dose of memantine (45 mg/kg), the Bmax (0.45 +/- 0.06) pmol/mg pro and Kd (22.88 +/- 4.42) nmol/L of NMDA receptor were significantly decreased (P < 0.01).

CONCLUSIONMemantine in certain dose range could protect against the down-regulation of NMDA receptor in rat brain, and alleviate organophosphorus poisoning symptoms to some extent. The recovery of AChE activity inhibition wasn't involved in the treatment with memantine on dichlorvos poisoning, therefore, atropine and a proper AChE reactivator (an oxime) should be used clinically.

Acetylcholinesterase ; metabolism ; Animals ; Brain ; drug effects ; metabolism ; Dichlorvos ; toxicity ; Dopamine Agents ; pharmacology ; Insecticides ; toxicity ; Male ; Memantine ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; metabolism