Aims: The aim of this study was to search for Bacillus thuringiensis (Bt) harbouring cry1A gene which could effectively control cotton pest, American bollworm, Helicoverpa armigera. Methods and Results: cry gene profiling of 50 Bt isolates showed the presence of cry1, cry2, cry3, cry4, cry7, cry8 and cry9 genes. None of the isolates harboured cry1 gene alone. It was always found in combination with cry3. There was no isolate positive for cry10 gene. Considering isolates with single cry genes, the frequency of cry4 was predominant (22%) followed cry2 (6%), cry3 (4%) and cry8 (2%). Isolates having two cry genes in combination had 14% incidence for cry2 + cry4, 12% for cry3 + cry4 and 10% for cry1 + cry3. The most dominant three gene linkage was cry1 + cry3 + cry4. Further profiling of cry1 gene showed that cry1K gene was abundantly present in all combinations such as cry1A, cry1D, cry1F and cry1I. However, cry1C existed independent of other subtypes. Finally, the Bt isolates with cry1A were analyzed for 16S rRNA gene, which showed two distinct groups of isolates on the basis of sequence homology. Bioassays of spore-crystal mixtures of 8, 17, 21 and 26 harbouring cry1 against neonate larvae of H. armigera showed LC 50 1288, 1202, 467AE7, 524AE8 and 108AE5 lg ml )1 . The SBS-Bt26 showed fourfold higher toxicity than the cry 1Ac harbouring positive control, HD-73. Conclusions: None of the isolates harboured single cry 1 gene. They were always in combination of two or three genes. A Bt isolate (Bt26) had fourfold higher toxicity against H. armigera larvae compared with the positive control HD 73 and hence can be commercially exploited to control insect pest. Significance and Impact of the Study: The inter relationship between the cry genes content and the toxicity may allow better understanding of Bt ecology.

The six most toxic Pakistani isolates of Bacillus thuringiensis (SBS Bt-23, 29, 34, 37, 45 and 47), which were previously characterized for their toxicity against larvae of mosquito, Anopheles stephensi, and the presence of cry4 gene, were used for cry11 (cry4D) gene amplification. A 1.9-kb DNA fragment of cry11 gene was PCR-amplified, cloned in expression vector pT7-7, and then used for transformation of E. coli BL21C. The optimum expression was obtained with 1 mM IPTG at 37 degrees C for 3 h. This gene showed different percentage homologies at protein level with scattered mutations in the toxic region. Biotoxicity assay of recombinant protein showed that Cry11 of SBS Bt 45 (DAB Bt 5) was the most toxic protein against third instar larvae of mosquito, A. stephensi, and has potentiality of a bioinsecticide against mosquitoes.

Probiotics are the beneficial microorganisms, catalase negative which restore microbial balance inside the gut of humans as well as animals. Lactobacillus the earliest probiotic that have the beneficial impact on health. These “Good Microorganisms” can be obtained not only from various non-dairy products but also from processed dairy products like. Another economically viable method is microencapsulation for preserving probiotics and the stability is improved by glucose. Even the vitamins manufacturer the probiotic bacterial agents. The health benefits of probiotics include increased immunological responses, relief of lactose intolerance symptoms, therapy for diarrhea, reduction in serum of cholesterol, production of vitamin, anticarcinogenic. Probiotics play a wide range in the host body (e.g., decreasing illnesses and stress, enhancing immunity, modulation of gut microbiota, nutritional assistance, improving quality of water, etc.). So, the positive effects of probiotics help to boost animal feed value and growth and improve aquaculture breeding and hatching rates. Probiotics can lower the prevalence and severity of illnesses, showing their promise to cure or prevent COVID-19. Lactobacillus casei also interact with epithelial cells with Toll-like receptors (TLRs) to improve the production of cytokines that are important in the enhancement of cell productivity and prevent apoptosis during restoration, which promote survival and proliferation. The preservation of the human GI or lung microbiota might help prevent COVID-19, as dysbiosis plays an essential role in people’s vulnerability to infectious illnesses. Most of the experimental studies proved that bacteria isolated from processed dairy products belonged to lactic acid bacteria and are declared as probiotic bacteria. In present review, various research studies regarding significance of probiotics as well as their extraction from processed dairy products are discussed.

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