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ThesisItem Open Access EFFECTS OF HEN EGG WHITE PROTEINS ON REPRODUCTION AND DEVELOPMENT OF RED FLOUR BEETLE (Tribolium castaneum)(PAU Ludhiana, 2010) Geetu Choudhary; R.K. ParshadThe red flour beetle, Tribolium castaneum (Herbst) (Coleoptera:Tenebrionidae) is a destructive pest of stored grains and it is renowned for adapting to various insecticides. The hen’s egg white proteins possess unique multifunctional properties including antimicrobial, antiviral, immunomodulatory and anticancer. Therefore, the present study was carried out to determine the insecticidal potential of hen’s egg white proteins to manage red flour beetle population. Protein samples obtained through salt fractionation (PPT-I and PPT-II) including whole egg white (WEW) were lyophilized. Different concentrations (2%, 3%, 4%, 5%, 10%) of each of the protein samples were tested separately and simultaneously by adding them to wheat flour and milk powder (75:25 w/w). The results indicated that mortality rate of the adult beetles was dependent on the type of treatment, concentration of protein samples and duration of feeding. WEW, PPT-I and PPT-II treatments delayed the emergence of developmental stages, reduced the fecundity rate, egg volume and reproductive effort of the female beetles. In multiple-choice feeding trials, beetles showed repellency towards the food containing PPT-I and PPT-II proteins. Marked abnormalities were observed in appearance and dimensions of the ovarioles and eggs which indicated that the hen’s egg white proteins caused considerable effect on the process of oogenesis. Scanning electron microscopic studies revealed changes in the structure of epithelial sheath cells of ovarioles probably caused due to resorption of ova in the beetles fed on PPT-II proteins. Higher resolution of the egg surface showed the presence of wrinkles or shrivelled appearance of the chorionic layer in PPT-I and PPT-II treated eggs and these deep folds and wrinkles were more conspicuous in the beetles fed on PPT-II proteins. PPT-II proteins appeared to be most effective among WEW, PPT-I and PPT-II samples in causing mortality and reduction of fecundity in T.castaneum.ThesisItem Open Access MUSCLE FATTY ACID PROFILES OF SOME WILD FISH SPECIES AND ENHANCEMENT OF OMEGA-3 FATTY ACIDS THROUGH DIETARY MANIPULATION IN Cyprinus carpio (Linn.)(PAU Ludhiana, 2012) Jyotika; H.S., SehgalThe study has been conducted on i) seasonal variations in muscle total lipid content (MTLC), fatty acid profiles and proximate composition of two wild fish species, Puntius conchonius (Hamilton-Buchanan) and P. sarana (Hamilton-Buchanan), to determine their nutritional status and ii) enhancement of omega-3 polyunsaturated fatty acids (PUFAs), considered vital for human health, through dietary manipulation in a cultured carp species, Cyprinus carpio (Linn.). Both the wild species were found to be rich in crude protein, which varied between 15.74% to 18.18% in P. conchonius and 15.15% to 18.04% in P. sarana during different seasons. Based on their MTLC (2.36% to 4.89%), both the species could be categorized as lean or low-fat fishes. However, the MTLC was higher and the moisture, ash and carbohydrate contents of the fishes were comparable with many other freshwater fish species. The fishes had good PUFA content. Omega-3 (n-3) PUFAs were 2.78% in P. sarana and 2.21% in P. conchonius. Total n-6 PUFAs were 4.56% in P. sarana and 4.20% in P .conchonius, and the values of n-3/n-6 ratio were 0.73 and 0.60 in P. sarana and P. conchonius, respectively. Linolenic acid formed 1.64% of the total n-3 PUFAs in P. sarana and 1.02% in P. conchonius. EPA constituted 0.31% in P. conchonius and 0.24% in P. sarana and DHA 0.75% in P. sarana and 0.69% in P.conchonius. Stearic acid was the dominant saturated fatty acid (SFA) forming 28.75% of the total SFAs in P. sarana and 28.10% in P. conchonius. The MTLC and n-3 PUFAs of C. carpio (Linn.), a cultured carp species could be enhanced through dietary manipulation. The MTLC could be increased from 1.44% in fish fed traditional diet with no flaxseed oil to 2.83% in fish fed 30% flaxseed oil diet. Similarly, the total n-3 PUFAs could be enhanced from 10.38% in fishes fed 0% flaxseed oil diet for 90 days to 18.67% in fishes fed 30% flaxseed oil diet for 60 days and 30% fish oil diet for 30 days.ThesisItem Open Access EVALUATION OF EUCALYPTUS AND CITRONELLA OILS AS REPELLENTS AGAINST HOUSE RAT, Rattus rattus (Linnaeus)(PAU Ludhiana, 2012) Ramandeep Kaur; Neena, SinglaExperiments were conducted in laboratory pens, maze and store houses using two essential oils namely eucalyptus and citronella oils to study their repellency against house rat, Rattus rattus. In laboratory pens, both oils were applied as paint, as encapsulated wax blocks and as spray each further with three modes of application i.e. daily, once a week, and alternatively in a week at three concentrations 5, 10 and 20%. Repellent effect of the two oils was observed at all the concentrations tested but significantly (P ≤ 0.05) higher effect of eucalyptus oil was in female rats at 20% concentration and of citronella oil in male rats at 10% concentration when applied as spray alternatively. Multichoice experiment conducted in T-maze using 5% eucalyptus and citronella oils as encapsulated wax blocks in choice with no treatment revealed significantly (P ≤ 0.05) higher repellent effect of eucalyptus oil compared to citronella oil in female rats. Bichoice experiment conducted in I-maze using 5% eucalyptus oil as encapsulated wax blocks in choice with no treatment showed significantly (P ≤ 0.05) reduced movements and activities of both male and female rats in treated zone compared to untreated zone. In store house experiments conducted using 5% eucalyptus and citronella oils as encapsulated wax blocks and as spray revealed more percent repellency and control success with eucalyptus oil as encapsulated wax blocks. Present studies thus reveal the efficacy of 5% eucalyptus oil encapsulated in wax blocks as repellent against R. rattus of both sexes.ThesisItem Open Access GASTROPOD INFESTATION IN PLANT NURSERIES AT DIFFERENT LOCATIONS AND THEIR MANAGEMENT WITH LATEX OF SOME PLANTS Thesis(PAU Ludhiana, 2011) HARJOT KAUR; Harjit Kaur)Survey of five plant nurseries at different locations viz. New Indian nursery and Gulrej nursery at Dhuri, Bhagwaan and Golden nursery at Malerkotla and Plant nursery PAU, Ludhiana for gastropod population revealed that one snail, Macrochlamys indica and one slug, Filicaulis alte Ferussac with three colour forms viz. black slug with yellow mid-dorsal streak, brown slug with black spots, black slug were infesting the plant nurseries. Slugs and snails preferred soil with low bulk density, high porosity and high moisture. Impact of three treatments on gastropod (snails and slugs) trapping was in following order- Damp conditions > Beer traps > Spots with Spinach. Snails and slugs excavated holes within leaves of ornamental plant saplings. More damage was observed during the months of June, July and October when the population density of snails and slugs active at soil surface was more. Hence, more emphasis should be laid down on gastropod control during the months of June, July and October to prevent the damage to ornamental plant saplings. Laboratory feeding tests revealed that out of the different concentrations of crude latex of different plants viz., Euphorbia splendens, Calotropis procera, Ficus bengalensis and Ficus elastica, bait with 2 per cent crude latex of E. splendens proved to be most effective bait resulting in 36.7 per cent mortality (F. alte, black slug with yellow mid-dorsal streak) with highest consumption among all the different concentrations of all the four plants. Thus, E. splendens latex acts not only as molluscicide but as attractant also.ThesisItem Open Access STUDY ON COMBINED EFFECT OF BROMADIOLONE AND CHOLECALCIFEROL (VITAMIN D3) AGAINST HOUSE RAT, Rattus rattus LINNAEUS(PAU Ludhiana, 2010) Navjot Kaur; Devinder Kaur, KocherCommensal rodents, especially house rat (Rattus rattus Linnaeus) causes extensive losses by feeding and contaminating the food products and also plays a role in spreading several diseases of health importance. House rats were trapped from various commensal situations and were fed for 5 days in no-choice on standard baits of bromadiolone (0.005%) and cholecalciferol (0.075%) and their combinations having different concentrations mixed in WSO-mix bait. Male and female house rats showed 81.30% and 95.80% average acceptability of standard bait of bromadiolone over plain bait respectively. Hundred per cent mortality of male house rats was observed within 3-6 days, while all female rats died on 4th day of feeding of 0.005% bromadiolone. Both the sexes showed a significant increase in blood clotting time (sec) after 48 hours of feeding as compared to that of 0 hour. The acceptability of standard bait of cholecalciferol (0.075%) over plain bait was 40.10% and 35.30% in male and female house rats, respectively and 100% mortality was observed within 4-10 days in male and 7-14 days in female house rats. Feeding of standard bait of cholecalciferol resulted in stop feeding action in the form of significantly less consumption of this bait from 3rd day onward. Serum calcium level (mg/dL) was found to be significantly high in 0.075% cholecalciferol fed male and female rats after 48 hours of its feeding as compared to 0 hour. R. rattus when fed on four formulated baits having different concentrations of bromadiolone and cholecalciferol i.e. combination-I (0.0025% bromadiolone+0.05% cholecalciferol), combination-II (0.001% bromadiolone+0.05% cholecalciferol), combination-III (0.0025% bromadiolone+0.01% cholecalciferol) and combination-IV (0.001% bromadiolone+0.01% cholecalciferol). Out of these four tested formulated baits, combination-IV (having the lowest concentration of bromadiolone and cholecalciferol) showed efficient rodenticidal potential because of synergistic effect of bromadiolone and cholecalciferol in this combination. It was able to produce 100% mortality and showed a significant delay in blood clotting time (sec) and rise in serum calcium level and was cost effective also. R. rattus L trapped from commensal situations of Ludhiana area showed no resistance towards standard bait of bromadiolone (0.005%) as both male and female house rats (n=60) showed 100% mortality.ThesisItem Open Access DEVELOPMENT OF ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) FOR DETECTION OF MELOIDOGYNE SPP. (ROOT-KNOT NEMATODE) ANTIGENS(PAU, 2010) Mandeep Kaur; Ghai, Jyotika KapurPlant parasitic nematodes are destructive pests worldwide that cause severe losses in agriculture. Root-knot nematodes (Meloidogyne spp.) are obligate, sedentary plant endoparasites that are extremely polyphagous in nature with a wide host range. Keeping in view the huge economic losses by this parasite, it is essential to control the disease at an early stage. For any control strategy to be effective, an early and accurate diagnosis is of paramount importance. Immunoassays have the inherent advantages of sensitivity and specificity; have the potential to identify and quantify these plant-parasitic nematodes. Hence, in the present studies, ELISA has been developed for the detection of Meloidogyne spp. antigens. This is based on detection of egg antigens, for which anti-Meloidogyne antibodies were produced by immunization of rabbits with soluble proteins of the eggs. The production of antibodies was confirmed by the appearance of precipitin lines in double immunodiffusion method. An indirect enzyme-linked immunosorbent assay (ELISA) was developed for detection and titration of these antibodies. Results indicated as high as 320 K titre of the antisera. Finally competitive inhibition ELISA was developed employing these anti-Meloidogyne antibodies for detection of Meloidogyne antigens. This is based on competition between solid phase bound antigens and free antigens for limited antibodies. Sensitivity of ELISA was 10 femtograms. This sensitivity was further enhanced to 1 femtogram with an additional step of pre-incubation, in which antibodies and free antigens are allowed to react before the competition. Competitive inhibition ELISA developed in the present studies has the potential of being used as an easy, rapid, specific and sensitive diagnostic tool for the detection of Meloidogyne infection.ThesisItem Restricted POPULATION DYNAMICS OF MOSQUITO LARVAE INHABITING STANDING WATER BODIES IN RELATION TO PHYSICOCHEMICAL PARAMETERS(Punjab Agricultural University, Ludhiana, 2013) DiptiMosquitoes breed in the aquatic systems which can either be man-made reservoirs or standing water bodies like ponds, swamps, marshes and paddy fields. These water bodies act as breeding grounds for mosquitoes, thus are responsible for the spread of mosquito borne diseases. So a survey was conducted to study the mosquito larvae population dynamics with an objective to find out their association with physico-chemical parameters of water in permanent and temporary standing water bodies. For this purpose water samples were collected at monthly intervals from four village ponds of Ludhiana district and paddy fields at PAU campus, Ludhiana. Mosquito larvae were enumerated from the different water samples and were identified under microscope with the help of standard morphological keys. Total mosquito larvae population i.e larval density (LD/L) and relative density of different types of mosquito larvae in terms of larval density index (LDI%) was calculated. Three types of mosquito larvae i.e Aedes, Anopheles and Culex were found and their LD was recorded to be maximum in rainy season at all the sampling sites. Culex was found to be the predominant type of larvae in water samples collected from all the ponds with percent LDI 62.7, 76.8, 56.6, and 87.6% respectively at Noorwala, Kasabad, Kaneja and Sujatwala villages as compared to other two genera of mosquito. Mosquito larvae population count was quite low in paddy fields as compared to village ponds. Estimation of different types of physical and chemical parameters from all the water samples collected from ponds and paddy fields revealed that mosquito larvae population was found to be positively correlated with temperature and alkalinity, while dissolved oxygen showed a negative association, other parameters tested were found to show non-significant correlation.ThesisItem Open Access IDENTIFICATION AND CHARACTERIZATION OF MELOIDOGYNE SPECIES-SPECIFIC PROTEINS(Punjab Agricultural University, Ludhiana, 2012) Navjot kaurRoot knot nematodes (RKNs) of genus Meloidogyne are among the most widely spread parasitic nematodes which attack a wide range of crops and limit their production. The egg masses of Meloidogyne nematodes were cultured on the tomato plants and were identified on the basis of perineal pattern of females by Scanning Electron Microscopy. Only single species of Meloidogyne i.e. M. incognita was identified, indicating that the maintained culture was pure. The crude homogenate/antigen was prepared from the egg masses of established pure culture by homogenization, sonication and centrifugation. Quantitative analysis of prepared homogenate showed an average protein content (mg/gm, wet weight of nematodes) of 128.43 ± 11.10. Protein profile of this crude protein extract by SDS-PAGE indicated a total of seven bands (five major and two minor) observed within a range of molecular weight 27 kDa to 118 kDa. Major band was observed between 50-90 kDa followed by a band between 36-50 kDa. Two minor bands were also observed between 27-36 kDa and 50-90 kDa. Healthy rabbits were immunized with crude protein/antigen of M. incognita egg masses and hyperimmune serum showed significantly higher levels (% of total protein) of Ig from 2.37±0.32 (0 DPI) to 6.98±1.06 (60 DPI), and CIC from 3.48±0.80 (0 DPI) to 12.13±0.56 (60 DPI). Double immunodiffusion technique indicated the production of antibodies, as precipitation line was observed due to the formation of lattices of antigen-antibody complexes. Western blotting resulted in the transfer of single band (lying between 50-90 kDa) from the protein profile of crude antigenic extract of M. incognita analysed on SDS-PAGE on the nitrocellulose membrane probed with anti-Meloidogyne antibodies, thus indicating the antigenic specificity of this proteinThesisItem Open Access STUDIES ON THE HABITAT, FEEDING AND BREEDING BIOLOGY OF BLACK KITE (Milvus migrans) IN AGROECOSYSTEM OF PUNJAB(Punjab Agricultural University, Ludhiana, 2013) Sachin KumarAgricultural land use is an important driver of biodiversity loss in developing countries. It was found that farming is the current and likely to be the future threat to globally threatened and near threatened bird species. The Black Kite (Milvus migrans) is a medium sized bird of prey in the family Accipitridae. The present study deals with habitat, feeding and breeding biology of Black Kite (Milvus migrans) in agroecosystem of Punjab. It was undertaken from December 2010 to May 2012 in 12 transects having different type of vegetation. Observations were taken in morning and evening twice a week regarding their habitat, feeding and breeding biology. It was found that Black Kite uses different types of areas for different activities like feeding, foraging, roosting and breeding. The number of territorial pairs of Black Kites increased steadily during this study but it was difficult to say whether this was caused by a real population increase or a simple population fluctuation. Most of the time, they were found in areas near human habitation. Feeding was maximally observed on garbage dumps. The other types of feeding materials were earthworms, rodents, reptiles, flesh and small birds. Flocking was also observed throughout the study periods but the size of flock varied in different areas. Nesting was mainly observed on Silver Oak followed Sapt Parni and Eucalyptus trees. The average nesting height was observed to be between 14.95 meters to 16.60 meters. The incubation period was between 30 to 35 days. The numbers of young ones varied from 1 to 3 with most of the nests having 2 young ones. In conclusion it was found that Black Kite feed on refuse garbage dumps, breeds on comparatively tall trees and plays an important role in cleaning the environment.