Partitioning the roles of insect and microbial enzymes in the metabolism of the insecticide imidacloprid in Drosophila melanogaster

Resistance to insecticides through enhanced metabolism is a worldwide problem. The Cyp6g1 gene of the vinegar fly, Drosophila melanogaster, is a paradigm for the study of metabolic resistance. Constitutive overexpression of this gene confers resistance to several chemical classes of insecticides, including the neonicotinoids exemplified by the insecticide imidacloprid (IMI). The metabolism of IMI in this species has been previously shown to yield oxidative and nitro-reduced metabolites. While levels of the oxidative metabolites are correlated with CYP6G1 expression, nitro-reduced metabolites are not, raising the question of how these metabolites are produced. Some IMI metabolites are known to be toxic, making their fate within the insect a second question of interest. These questions have been addressed by coupling the genetic tools of gene overexpression and CRISPR gene knock-out with the sensitive mass spectrometric technique, the Twin-Ion Method (TIM). Analysing axenic larvae indicated that microbes living within D. melanogaster are largely responsible for the production of the nitro-reduced metabolites. Knock-out of Cyp6g1 revealed functional redundancy, with some metabolites produced by CYP6G1 still detected. IMI metabolism was shown to produce toxic products that are not further metabolized but readily excreted, even when produced in the Central Nervous System (CNS), highlighting the significance of transport and excretion in metabolic resistance.

A novel method for quantification of lactose in mammalian milk through HPTLC and determination by a mass spectrometric technique

A novel HPTLC method was used to estimate the lactose concentration present in the milk of different mammaliansviz: Indian mare (Equus caballus), Cow (Bos primigenius indicus), Buffalo (Bubalus bubalis), Camel (Camelus dromedarius) and Donkey (Equss asinus).