Insecticide resistance and malathion carboxylesterase in the sheep blowfly,Lucilia cuprina

1994 ◽  
Vol 32 (1-2) ◽  
pp. 9-24 ◽  
Author(s):  
Steven Whyard ◽  
Robyn J. Russell ◽  
Virginia K. Walker
Keyword(s):  
Insecticide Resistance ◽  
Lucilia Cuprina ◽  
Malathion Carboxylesterase

A method for the detection and measurement of insecticide resistance in larvae of Lucilia cuprina (Wied.) (Dipt., Calliphoridae)

1973 ◽  
Vol 63 (1) ◽  
pp. 99-102 ◽  
Author(s):  
N. A. Roxburgh ◽  
G. J. Shanahan
Keyword(s):  
Blood Serum ◽  
Insecticide Resistance ◽  
Lucilia Cuprina ◽  
Resistance Factors ◽  
First Instar ◽  
Resistant Strains

AbstractIn this method the toxicant is absorbed with blood serum onto a loose roll of chromatography paper; first-instar larvae of Lucilia cuprina (Wied.) are inserted into the air spaces of the roll and allowed to feed. Mortality of the larvae is assessed 24 h later. Typical results using diazinon, dichlofenthian and chlorfenvinphos against susceptible and resistant strains of L. cuprina showed larvae to have resistance factors 6–8 times higher than those for adults determined by established techniques.

scholarly journals Proteomic Analysis of Glutathione Transferases from Lucilia Cuprina

2021 ◽  
Author(s):  
◽  
Ramavati Pal
Keyword(s):  
Insecticide Resistance ◽  
Proteomic Analysis ◽  
Developmental Stages ◽  
Adult Stage ◽  
Glutathione Transferases ◽  
Lucilia Cuprina ◽  
Main Body ◽  
Post Translational Modification ◽  
2D Page ◽  
Gst Activity

<p>The glutathione transferases are a family of multifunctional enzymes involved in detoxification of xenobiotic and endogenous electrophilic compounds. Interest in insect GSTs has primarily focused on their role in insecticide resistance. The sheep blowfly, Lucilia cuprina is a major economic problem for the sheep meat and wool industries in Australasia and hence this thesis has attempted the study of the Lucilia cuprina GST family, using proteomics, with a view to eventually determining their role in insecticide resistance. Combinations of different affinity matrices (glutathione-Sepharose matrix (GSH) followed by dinitrophenyl-glutathione-Sepharose matrix (DNP-GSH)) and two-dimensional electrophoresis has successfully isolated members from major four insect GST classes: Sigma, Delta, Epsilon and Omega. Drosophila melanogaster has been used as a model insect throughout as a basis for comparison. To characterise Lucilia GSTs, the whole metazoan fragmentation database was used for sequence alignment with Lucilia peptides. This approach is broad and speculative but predicts a possible classification of the GSTs based on % similarity and % identity. This method of characterisation yielded match scores that provided a basis for classification, which must at present be regarded as tentative and in need of confirmation. In D. melanogaster and L. cuprina, GSH affinity-purified extracts showed the presence of only Sigma and Delta GSTs. In D. melanogaster, the DNP-GSH affinity-purified GSTs showed mostly the presence of Epsilon and Omega GSTs whereas in L. cuprina no Omega GSTs were detected. In both species, the migration pattern of Delta GST on 2D PAGE gel indicated possible post-translational modification. The results from analysis of LC-MS/MS data by the software PEAKS suggested deamidation at asparagine and glutamine residues in a limited number of the matched peptides of Delta GST. GST activity was present in all developmental stages of L. cuprina. The number of isoenzymes and their extent of expression vary as the insect develops. Delta GSTs were present in all developmental stages. The Sigma GST started expressing from the larval stage and was abundantly present in adult stage. The DNP-GSH affinity matrix purified GSTs which have been tentatively classified as Mu-like GSTs were present in egg, larvae and pupae but totally absent in adult stage. The GST families were characterised by proteomics in the main body sections of L. cuprina. Higher GST activity towards 1-chloro-2, 4-dinitrobenzene (CDNB) was found in the thorax (65.2 %) followed by the abdomen (19.6%) and the head (15.2%). The cytosolic GSTs of a resistant strain (PY81) of L. cuprina had significantly higher (2.26- and 2.6- fold) activity than the susceptible strains (NSW and CSIRO) towards CDNB and 2, 3-dichloro, 4-nitrobenzene (DCNB) respectively. The proteomic analysis of DNP-GSH purified extract from susceptible and resistant strains showed quantitatively higher expression of GSTs on 2D PAGE gel of the PY81 strain. The in vitro interaction of purified GSTs and model insecticides studied by high performance liquid chromatography revealed that Delta and DNP-GSH affinity-purified GSTs catalyse the conjugation of the insecticides to reduced glutathione but Sigma GST had almost no activity.</p>

scholarly journals Field Studies on Insecticide Resistance in the Australian Sheep Blowfly, Lucilia Cuprina

1980 ◽  
Vol 33 (6) ◽  
pp. 725 ◽  
Author(s):  
MJ Whitten ◽  
JM Dearn ◽  
JA McKenzie
Keyword(s):  
Insecticide Resistance ◽  
Larval Development ◽  
Field Studies ◽  
Lucilia Cuprina ◽  
Insecticide Residue ◽  
Genotype Frequencies ◽  
Australian Sheep ◽  
Free Environment ◽  
Eggs And Larvae ◽  
Oviposition Sites

Eggs and larvae of L. cuprina were coIJected from natural fly strikes in a flock of Merino ewes in which sheep had been either treated with the insecticide dieldrin or left as controls. An analysis of gene and genotype frequencies of Rdl locus, which determines resistance to dieldrin, provides support for the existence of strong selection operating during larval development on sheep whose fleece contain insecticide residue. Resistance genotypes appear to be at a disadvantage both in the laboratory and in the insecticide-free environment of control sheep. There is no evidence that flies of different resistance status choose oviposition sites on the basis of the presence of dieldrin residues in the fleece.

scholarly journals Estimation of the Relative Viabilities of Insecticide Resistance Genotypes of the Australian Sheep Blowfly Lucilia cuprina

1984 ◽  
Vol 37 (2) ◽  
pp. 45 ◽  
Author(s):  
JA McKenzie ◽  
MJ Whitten
Keyword(s):  
Insecticide Resistance ◽  
Organophosphorus Compound ◽  
Organochlorine Compound ◽  
Lucilia Cuprina ◽  
Australian Sheep

Artificial strikes were initiated on sheep at different times after treatment with either diazinon (an organophosphorus compound) or lindane (an organochlorine compound).

scholarly journals Proteomic Analysis of Glutathione Transferases from Lucilia Cuprina

2021 ◽  
Author(s):  
◽  
Ramavati Pal
Keyword(s):  
Insecticide Resistance ◽  
Proteomic Analysis ◽  
Developmental Stages ◽  
Adult Stage ◽  
Glutathione Transferases ◽  
Lucilia Cuprina ◽  
Main Body ◽  
Post Translational Modification ◽  
2D Page ◽  
Gst Activity

<p>The glutathione transferases are a family of multifunctional enzymes involved in detoxification of xenobiotic and endogenous electrophilic compounds. Interest in insect GSTs has primarily focused on their role in insecticide resistance. The sheep blowfly, Lucilia cuprina is a major economic problem for the sheep meat and wool industries in Australasia and hence this thesis has attempted the study of the Lucilia cuprina GST family, using proteomics, with a view to eventually determining their role in insecticide resistance. Combinations of different affinity matrices (glutathione-Sepharose matrix (GSH) followed by dinitrophenyl-glutathione-Sepharose matrix (DNP-GSH)) and two-dimensional electrophoresis has successfully isolated members from major four insect GST classes: Sigma, Delta, Epsilon and Omega. Drosophila melanogaster has been used as a model insect throughout as a basis for comparison. To characterise Lucilia GSTs, the whole metazoan fragmentation database was used for sequence alignment with Lucilia peptides. This approach is broad and speculative but predicts a possible classification of the GSTs based on % similarity and % identity. This method of characterisation yielded match scores that provided a basis for classification, which must at present be regarded as tentative and in need of confirmation. In D. melanogaster and L. cuprina, GSH affinity-purified extracts showed the presence of only Sigma and Delta GSTs. In D. melanogaster, the DNP-GSH affinity-purified GSTs showed mostly the presence of Epsilon and Omega GSTs whereas in L. cuprina no Omega GSTs were detected. In both species, the migration pattern of Delta GST on 2D PAGE gel indicated possible post-translational modification. The results from analysis of LC-MS/MS data by the software PEAKS suggested deamidation at asparagine and glutamine residues in a limited number of the matched peptides of Delta GST. GST activity was present in all developmental stages of L. cuprina. The number of isoenzymes and their extent of expression vary as the insect develops. Delta GSTs were present in all developmental stages. The Sigma GST started expressing from the larval stage and was abundantly present in adult stage. The DNP-GSH affinity matrix purified GSTs which have been tentatively classified as Mu-like GSTs were present in egg, larvae and pupae but totally absent in adult stage. The GST families were characterised by proteomics in the main body sections of L. cuprina. Higher GST activity towards 1-chloro-2, 4-dinitrobenzene (CDNB) was found in the thorax (65.2 %) followed by the abdomen (19.6%) and the head (15.2%). The cytosolic GSTs of a resistant strain (PY81) of L. cuprina had significantly higher (2.26- and 2.6- fold) activity than the susceptible strains (NSW and CSIRO) towards CDNB and 2, 3-dichloro, 4-nitrobenzene (DCNB) respectively. The proteomic analysis of DNP-GSH purified extract from susceptible and resistant strains showed quantitatively higher expression of GSTs on 2D PAGE gel of the PY81 strain. The in vitro interaction of purified GSTs and model insecticides studied by high performance liquid chromatography revealed that Delta and DNP-GSH affinity-purified GSTs catalyse the conjugation of the insecticides to reduced glutathione but Sigma GST had almost no activity.</p>

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