Development of Species-specific Polymerase Chain Reaction (PCR) Targeting on Mitochondrial D-loop for Identification of Buffalo and Goat Raw Meat

Abstract: Today's, consumers are concerned about the meat they eat and also demandaccurate labeling. Mitochondrial Analysis of DNA was the most frequently used DNA, becauseof its highly conserved sequences in various organism species. In this study, a rapid,reproducible and simple method for simultaneous identification of multiple meat species ina single step mitochondrial DNA based test has been developed based on the designing ofspecies-specific primer. Meat samples of goat and buffalo were selected to verify theapplicability of the technique. A species specific forward and reverse primer was designedwith the help of the primer3 tool for amplification of mitochondrial D-loop region. Thespecies-specific primers were verified in silico by SnapGene software. The two pairs ofprimers amplified the expected fragment of 338bp for buffalo and 450bp for goat. The changein the size of the PCR product was due to the existence of highly polymorphic regions withinthe buffalo and goat D-loop region. The tested species gives a unique band pattern for eachspecies by using successful amplification of these polymorphic regions in the D-loop region.Overall, the simplicity of amplification of mitochondrial D-loop region could make thistechnique suitable for meat authentication in routine analysis.

Seasonal dynamics and management of whitefly ( Bemesia tabaci Genn.) in tomato ( Solanum esculentum Mill.)

ABSTRACT Studies on seasonal dynamics of white fly (Bemesia tabaci) on tomato (Solanum esculentum var. Pusa Ruby) revealed that it appeared first during the 13th and reached maximum during 21st standard meteorological week. A positive correlation between adult population and abiotic factors viz. temperature (maximum and minimum) and sunshine hours was observed, whereas humidity (maximum and minimum) and rainfall showed a negative correlation with it. Taken together, the key weather parameters studied, caused 89.00 per cent variation in whitefly population (R2 value). Combination of carbofuran (soil application) + imidacloprid (seed treatment ) + imidacloprid (foliar application) proved significantly superior and caused maximum reduction in whitefly population followed by imidacloprid (seed treatment) + thiomethoxam (spray), imidacloprid (seed treatment) + imidacloprid (spray), imidacloprid (seed treatment) + dimetheoate (spray), carbofuran (soil application) + malathion (spray), and imidacloprid (seed treatment) + yellow sticky traps. The highest cost benefit ratio of 1:25.04 was recorded in case of carbofuran (soil application) + imidacloprid (seed treatment ) + imidacloprid (foliar application) followed by 1:22.38 for imidacloprid (seed treatment) + thiomethoxam (spray) ; 1:21.81 for imidacloprid (seed treatment) + imidacloprid (spray); 1:19.27 imidacloprid (seed treatment) + dimetheoate (spray); 1:19.48 carbofuran (soil application) + malathion (spray), and 1:8.33 for imidacloprid (seed treatment) + yellow sticky traps. The soil application of carbofuran + seed treatment with imidacloprid and three foliar sprays of imidacloprid at fortnight interval starting 40 days after transplanting is found effective and is advised for whitefly management in susceptible tomato cultivars.