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ThesisItem Open Access Detection and quantification of chlorpyrifos and endosulphan residues in dead animal’s using high performance liquid chromatography(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2007-05) Tyagi, Amit; Dixit, V.P.In the present study, about 168 tissue samples were analyzed for the presence of endosulfan and chlorpyrifos residues and 74 for presence of monocrotophos using the standardized method. The residues were extracted by treating with acetonitirile followed by liquid-liquid partition with sodium sulfate solution (2.5%): dichloromethane. The extracts obtained after dehydration on sodium sulfate column were cleaned up by performing adsorption chromatography on alumina column. The detection and quantification of these residues was carried out with the help of High Performance Liquid Chromatography using Diode Array Detector. Five (7.14 %) out of 70 muscle, 2 (2.85 %) of 70 liver and none of 70 kidney tissues were detected positive for endosulfan α residues. Out of these none of the samples violated the prescribed limits given by CODEX. Only one (1.42 %) of muscle tissue sample out of 70 was detected positive for endosulfan β residues. Out of these also none of the samples violated the prescribed limits given by CODEX. Seven (10.00 %) of muscle tissue, 6 (8.57 %) of kidney and 2 (2.85 %) of liver were detected positive for endosulfan sulfate residues. Out of these total samples none of the samples violated the prescribed limits given by CODEX. Chlorpyrifos residues were detected in 5 (7.14 %) of 70 muscle samples, 4 (5.71 %) of 70 liver and none of kidney tissue was detected positive. None of these samples also violated the prescribed limits given by CODEX. All of the 121 samples analyzed for presence of monocrotophos were found negative.ThesisItem Open Access Physico-chemical and microbiological characteristics of Ganges water from Gangotri (Uttarakhand) to Kanpur (Uttar Pradesh)(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2008-05) Singh, Yogesh Pratap; Dixit, V.P.The present study was conducted to know the pollution load in Ganges water. A total of 108 water samples were collected from 27 different places between Gangotri (Uttarakhand) and Kanpur (Uttar Pradesh). Samples were analyzed for their different physico – chemical and microbiological characteristics. The whole study area was divided into three groups viz. A, B.and C. Group A included the places that exclusively come under hill areas from Gangotri to Tehri, Group B included 11 places, starting from Laxaman Jhula (Rishikesh) to Garh Mukteshwar. Places of group C included those places which were well known for their pollution load like Kanpur. During the present study it was observed that the level of pollution was maximum in places of group C. Places of group B were slightly less polluted and least pollution was observed in places of group A. Values of physico– chemical parameters were very high than the normal values in groups. Microbial counts were also on increased level which showed that proper water treatment should adopted before using the Ganges water as drinking purpose. Physico–chemical and microbial changes were less in places of group A but it was not as low as it can be used for drinking purpose according to the standard laid down by ICMR, ISO and WHO with regard to physico–chemical and microbiological characteristics. By preventing further increase in pollution level and establishing water treatment plants we can try to reduce the pollution level in Ganges water.ThesisItem Open Access Molecular identification of animal species using Polymerase Chain Reaction based techniques(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2008-06) Karabasanavar, Nagappa; Singh, S.P.Species-specific PCR assays were developed for the authentic identification of cattle, buffalo, sheep, goat, pig and chicken. The mitochondrial D-loop was targeted in cattle, buffalo, sheep, goat, and pig; while nuclear 5-aminolevulinate synthase gene was used as target for amplification in chicken. The developed species-specific PCR assays yielded products specific to the species studied. In order to exclude the possibility of cross amplification, 25 animal species including different breeds were considered that consisted of mammals, birds, rodent and fish species covering most of the domestic, pet and wild animals. The suitability of the species-specific PCR assays was confirmed in raw (n=20), cooked (60,80 and 100oC for 30 min), autoclaved (121oC for 30 min) and the micro-oven processed meat and meat products such as kabab, mutton curry, chevon curry, pork sausage, chicken patties, chicken products (samosa, nuggets and loaves). The sensitivity of the PCR was established to be at 0.1% in all species studied for the detection of adulteration and the limit of detection was 0.1 pg in cattle and goat, 1 pg in sheep and 10 pg in buffalo, pig and chicken. Based on the present investigation it was concluded that, the species specific PCR assays developed in this study could be used for the authentication of raw, cooked (up to 121oC) and adulterated (up to 0.1%) animal tissues and their products for the specific identification of cattle, buffalo, sheep, goat, pig and chicken. For the simultaneous detection of cattle and buffalo a multiplex PCR was developed using species-specific forward and common reverse primers. Further, for the identification of an unknown species an alternative approach was used, that involvedisolation of total DNA from the cells, PCR amplification of a region of mitochondrial 12S rRNA gene, sequencing of the PCR amplicon and analysis of the sequence. Using this approach, nine species of animals (leopard, Bengal and Siberian tigers, goral, muntjak, sika deer, camel, parakeet and a black kite) were unambiguously identified. This research work presents novel diagnostic primers for 6 animal species and validates the sequence analysis as a tool for identification animal species.