Graduate Thesis Or Dissertation
 

Carboxylesterases in the house fly (Musca domestica, L.), flesh fly (Sarcophaga bullata, Park.), and black blow fly (Phormia regina, Meig.)

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https://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/2f75rc08n

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  • The carboxylesterases of three species of diptera, the flesh fly (Sarcophaga bullata, Park.), the black blow fly (Phormia regina, Meig.), and the house fly (Musca domestica, L.) were compared in relation to hydrolytic activity against three substrates, response to inhibition and induction, isozyme composition, and age-dependent variation in activity. Similar studies were also conducted with an insecticide-resistant house fly strain. Most of the experiments were with adult female flies but male flies as well as larvae and pupae were included in several experiments. Naphthyl acetate was the substrate common to all experiments and the esterases hydrolyzing this compound were further characterized according to their inhibition by eserine, parahydroxymecuribenzoate (PHNB), and paraoxon. The hydrolytic activity observed in the presence of eserine and PHNB or blocked by paraoxon was considered to be due to carboxylesterases of the B-type. Two other esters, hydroprene, an insect growth regulator similar in structure to the natural juvenile hormone, and juvenile hormone-1, were also used as substrates. Neither the hemolymph nor the cytosol esterases of the three species were susceptible to eserine inhibition, indicating that the samples contained minor quantities of the choline esterases. PHNB had little inhibitory effect on the flesh fly esterases of both hemolymph and soluble fraction. However, this inhibitor reduced the activity of the blow fly esterases by more than 50 percent, indicating a high content of aryl esterases in this species. The house fly esterases were also inhibited by PHNB but to a less extent than those of the blow fly. As expected, paraoxon was a strong inhibitor of the esterases in both hemolymph and soluble fraction of the three species. The total enzyme content of the flies was estimated from the hemolymph volumes determined earlier and the activity per unit volume found in the age-dependency experiments. The daily average activity estimated in this way was highest in the flesh fly, about four times that of the blow fly, and lowest in the house fly, about one-seventh that of the flesh fly. The values were even lower in the case of the resistant house flies, about 1/40th those of the flesh fly. When hydroprene was used as a substrate in the age-dependency experiments, the activity profiles were similar to those found with naphthyl acetate as substrate indicating that the same enzymes were involved in the hydrolysis of this JH analogue. Of the three species, the flesh fly esterases were most active against hydroprene and the blow fly and resistant house fly enzymes were the least active. The flesh fly hemolymph enzymes averaged, on a per-day basis, up to three times more hydroprene cleaving activity than those of the resistant house flies. The electrophorese experiments with hemolymph and cytosol revealed the presence of several carboxylesterase isozymes. As many as 14 different esterase-active bands were found in flesh fly hemolymph, 11 in blow fly hemolymph, and 12 in house fly hemolymph. Similar results were obtained with the cytosol. These bands were placed in five groups according to their mobilities in the poly acrylamide gels and it was evident that the isozymes migrating to the midpoint of the gels were responsible for most of the carboxylesterase activity. Some slow-moving isozymes were also thought to be responsible for this type of activity. Other evidence obtained during the electrophoreses experiments indicated that the isozymes hydrolizing hydroprene were from the same groups as those cleaving naphthyl-acetate while those which attacked JH-1 had different electrophoretic characteristics. Both hydroprene and naphthyl acetate esterases were induced by topically applied hydroprene. The response was dose dependent in the range 1-10 ug for flesh flies and 1-5 ug for house flies and blow flies. However, the response was variable and appeared to depend on the age of the flies at the time of the treatment. Increases in esterase activity ranged from 1.5 to 4-fold with the flesh fly enzymes being the most responsive and those of the blow fly the least. The possibility of JH-1 esterase induction by hydroprene was not investigated.
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