Pathogenic Escherichia coli is found in the contaminated raw meat, raw milk, raw vegetables, contaminated water. The present work was based on the entomopathogenic effect of E.coli against stored grain pest Tribolium castaneum. Microbial screening of pathogenic bacteria was done by using different media such as TSB media, MacConkey media and Novobiocin media. Rearing of Tribolium castaneum was done and different stages of Tribolium castaneum such as larvae, pupae and adult were observed. Bioassay technique was applied for the detection of effect of E.coli against the Tribolium castaneum. The Bioassay was applied in two groups e.g. Control group and Experimental group. The pellet of grown culture of E.coli sample was mixed in the diet of Tribolium castaneum. The effect of E.coli was observed against 2nd Instar larvae of Tribolium castaneum upto three days and seven days.The mortality rate in both control group and Experimental group was approximately 10%. The results had shown that E.coli did not have any effect against Tribolium castaneum."

Pathogenic Escherichia coli are found in the contaminated raw meat. Contamination is common in Pakistan due to unhygienic condition. The present work was based on the entomopathogenic effect of E.coli against Tribolium castaneum. The cultured sample of E.coli O157 isolated from raw meat was used in this study. Microbial screening of pathogenic bacteria was done by using different media such as TSB, MacConkey and Novobiocin. Rearing of Tribolium castaneum was done in the Laboratory of LCWU. Bioassay technique was applied for the detection of effect of E.coli against the Tribolium castaneum. The Bioassay was applied in triplicates in bioassay glass vials. The results had shown that E.coli did not have any effect against Tribolium castaneum.

Microbial control of insect pest of crops using entomopathogens is pest management strategy. Bacillus thuringiensis has a great potential for this purpose. The focus of the present study was to determine the individual and combined effect of different insecticides such as Saraca indica as a botanical insecticide, B. thuringiensis and chemical insecticides against larvae and adults of T. castaneum. Results were analyzed by SPSS probit analysis, LC50 was noted for all these insecticides and results were compared with each other. According to the results from the individual insecticide B. thuringiensis show high mortality with harmless effect to environment. Synergistic effect with high mortality rate was observed at 24 to 48 h, for both larvae and adults of T. castaneum. From the study it was observed that mortality rate increases as the concentrations increases, and larvae show higher mortality because they were more susceptible to these insecticides as compared to adults. Percentage mortality of T. castaneum in combined bioassay sodium citrate, S. indica, B. thuringiensis and Bifenthrin+cypermethrin was compared with the mortality of individual insecticides. 61% larvae & 55% adults

The red flour beetle (Tribolium castaneum) is a common pest insect known for attacking and infesting stored flour and grain. Bacillus thuringiensis (B.t) is a Gram positive, soil dwelling bacteria commonly used as a biological alternative to a pesticide. The present project was designed for the study of combined insecticidal effect of B. t along with boric acid, insecticide cypermethrin and the plant leaves powder, that is, Azadirachta indica against the T. castaneum to check the mortality rate of these insects and LC50 was calculated. Bacteria were isolated from rich soil, pulse dust, grain dust and its growth was checked on the T3 media by applying the microbial techniques. The results obtained show synergism when applied in combined concentration of B.t, boric acid, cypermethrin and powder of neem leaves in 1 g of diet. The combined concentration of each gave higher mortality to larvae and the percentage mortality was calculated as 33, 16 and 16% after 24 h, 41, 25 and 16% after 48 h and 50, 25 and 16% after 72 h in three different combined concentrations as compared to individual concentration. Likewise, in case of adults the percentage mortality was 33, 16 and 13% after 24 h,

Bacillus thuringiensis spore forming, Gram positive aerobic bacteria whose spores consist of the crystal protein which are found to have the insecticidal activity to the specific insect orders. Bacillus thuringiensis are considered to be cosmopolitan in nature and a great difference occurs between the two strains from different habitat. The present study was conducted on the collection of samples and solation of the B.t. from these samples. These isolates were identified on the basis of microscopic techniques and staining techniques. Culturing of these isolates was made on the specific media such as T3. Culturing of these bacteria was done to get enough amount of the B.t. spore cell. Rearing of the Tribolium castaneum a flour beetle was done by understanding the physiology of the beetles. Then biotoxic effect of these bacteria was determined by the process of Biotoxicity assay of the Bacillus thuringiensis spores against Triboium castaneum in glass vials in triplicates. The seven day repeated observation of these isolates was presented in the tabular form and LC50 is calculated by the Probit analysis programme.

Red flour beetle is one of the major insect pests of stored grain product in warm and tropical regions of the world. It is the most abundant and injurious pest in flour mills, bulks of grains, oil seeds, stored cereals and warehousing facilities. The life cycle requires 7 to 12 weeks. Bacillus thuringiensis (Bt) is a biological insecticide that makes it useful for pest control. It is naturally occurring bacterium common in soil throughout the world. B. thuringiensis acts by producing proteins (toxic crystal) that react with the cells of the gut lining of susceptible insects. Bacillus thuringiensis affected insects generally die from starvation, as the bacteria enter the insect's blood and reproduce within the insect; it is the reaction of the protein crystal that is lethal to the insect. The development of environment friendly insecticides like DDVP+Tetramethrin, having specificity to insects along with low toxicity to vertebrates, has captured worldwide attention of entomologists. Chemical like potassium chloride is very effective in controlling insect pests. Insecticidal plants have replaced them as they are more effective in crop rotation.

The study conducted on the toxic effect of the Bacillus thuringiensis spores against the Tribolium castaneum can be of great importance in eliminating the stored pest grains. There are a lot of future perspectives of this study which are given below: Proper processes for the collection and handling of the samples should be under consideration in order to avoid the incorrect and contaminated samples from different sources. According to the nutritional requirements of the Bacillus thuringiensis a specific solid media called as T3 media is used for the culturing of these bacteria and incubation period of these is 72 hours for spore formation. To understand the physiology of the Tribolium castaneum is quite helpful in understanding the rearing processes of these beetles. Biotoxiciy assay provides a reliable method for determining the susceptibility of the Tribolium castaneum against Bacillus thuringiensis that provide an easy method to determine the mortality ratio. The spores of the Bacillus thuringiensis can be of great importance in the agriculture in reducing the considerable population of the stored grain pest such as Tribolium castaneum and also some other pests.

A detailed study was undertaken in order to determine the safe dose of mustard seed meal in the diet for Japanese quails which can be used at the production level without having deleterious effects on the growth of this bird. For this purpose, some 1500 birds were fed different levels of mustard seed meal in isocaloric and isonitrogenous diets. The results of feeding of these diets for 30 days show that there are no toxic effects on the growth of birds. In fact the birds on the experimental rations were either comparable or heavier than the control except 5% group. Different biochemical and physiological studies were performed on the liver show that glucosinolates present in the mustard did not have the drastic effects which have been reported in the literature for mammals and other domestic birds. The changes reported herein were seemed to have more of the manifestation of the age rather than the effect of glucosinolates.

Bacillus thuringiensis cry proteins are effective in controlling harmful insects. However, the mechanism of Bt protein pesticidal action is not well understood. It is assumed that the pesticidal protein has affinity for specific receptors in the midgut of the susceptible larvae and binds irreversibly to create holes in the gut leading to eventual death of the target larvae. Main aim of the research is to identify, purify and characterize Bt delta endotoxin receptor in important pest American bollworm (Helicoverpa armigera). The study presented in this thesis is endeavored to; (1) discovery of a receptor protein, (2) purification of the receptor, (3)cloning of the newly discovered protein gene in E.coli, (4) expression studies on recombinant protein, (5) Complete sequencing of the cloned gene. The structural gene was cloned in expression vector. The cloned gene was sequenced and studied expression. The cloned, expressed, purified receptor protein was found to have characteristics of native protein of H.armigera. A new method for quick detection of Bt-receptor in crude BBMVs extract, was established.The sequence has novelty and Accession # A59445)or Q7M4K6.

PMA minority of Escherichia coli (E. coli) strains are capable of causing human illness. E. coli O157:H7 has now been reported from Pakistan. Infection with E. coli O157:H7 may present with a wide range of clinical manifestations. This research work was based on Microbial, Immunological screening and Molecular study of pathogenic bacteria E. coli in meat samples (Beef, Mutton, Quail and Fish), in hawkers foods:dahi bhala, fruit chat, fruit juices and milk shakes. 300 Meat samples, fruit juices and milk shakes were collected from Lahore. 300 Dahi bhala & fruit chat samples were collected from Lahore and Gujranwala. Different media & mEC with Novobiocin,used for Growth,identification and selective isolation of E. coli O157:H7. 225 meat, 147 Dahi bhala and 146 fruit chat samples were identified as contaminated with E. coli by culture method. Fresh juices and milk shakes are also found contaminated.Production of anti-E. coli was performed & used for Immunological studies of all samples, verified the results by using commercial mAb.Molecular studies of E. coli O157:H7 were performed.Crude proteins from E. coli were extracted & band patterns were observed by SDS-PAG

A cry gene, obtained from a locally isolated Bt. strain, CAMB # 30382, highly effective against Tribolium castenium, was amplified through PCR. The gene was ligated to a cloning vector pGEM-T and was cloned into DH5 . Its characterization showed homology with coleopteran-specific cry3f gene from Bt. strain buibui, as judged by Southern hybridization, PCR, and Restriction analysis. The alignment of the sequenced gene with gene sequences present in gene data bank showed two silent mutations, at nucleotide 676 and 701. However, two other nucleotide changes at position 2887 and 2902 of holotype cry3f gene were resulting from amino acid change near 3 end were observed. These changes were causing change in the stretch of amino acids from valine, tyrosine, serine, glutamine & isoleucine in the holotype cry3f to glycine, phenylalanine, alanine, asparginine respectively in the clone from CAMB isolate 30382. This change may have drastic effect on the toxicity spectrum of the cry protein isolated from this local isolate in comparison with the cry protein from the Buibui. serovar japonesis. Partial sequencing has shown it to be a varient of cry3 gene, which may be a novel cry gene.

More than 700 serotypes of E. coli have been identified. The different E. coli serotypes are distinguished by their "O" and "H" antigens on their bodies and flagella, respectively. The E. coli serotypes that are responsible for the numerous reports of contaminated foods and beverages are those that produce Shiga toxin, E. coli O157: H7 was first recognized as a food borne pathogen in 1982 during an investigation into an outbreak of hemorrhagic colitis (bloody diarrhea) associated with consumption of contaminated hamburgers. Present work was based on bacterial contamination of food samples that is very common due to unhygienic conditions, . Undoubtedly in past 10-20 years the impact of food borne diseases has been over whelming in Pakistan. Developed detection methods can detect more than 10,000 pathogenic E.coli in food and water samples while ten pathogenic E.coli are enough to cause human infections like diarrhea and other complicated diseases. Microbial screening and Immunological studies (Dot Blot assay) of samples was done. The purpose of this study is to develop awareness about pathogenic E.coli and importance of its timely detectio