Biotechnology and Bioforensics

The main objective of this work is amplification and sequence analysis of TPI gene which is a structural gene of operon from Lactobacillus delbrueckii. The cow milk was collected under good conditions and analyzed for bacterial load. Many strains of lactic acid bacteria were obtained, out of which 3 strains were homofermentative lactobacilli. Different biochemical tests were done for the identification of Lactobacillus delbrueckii. The TPI gene of Lactobacillus delbrueckii is involved in the maximum production of bacteriocin. Genomic DNA was isolated from Lactobacillus delbrueckii and its TPI gene was successfully amplified using polymerase chain reaction. Further, sequence analysis of TPI gene was done using the bioinformatics tools and the phylogenetic study was made. From the sequence analysis we can infer that the TPI gene sequence of Lactobacillus delbrueckii has close resemblance with the Escherichia coli sequence.

In this era Bacteria are mostly considered as pathogenic. Bacteria though are pathogenic but still a few are friendly and essential for human growth and immunity; called as Probiotics. Lactobacillus model moiety is one of such probiotic bacterium which when introduced in sufficient colony serves as new prophylaxis and curing agent. This will increase the innate immunity of humans. Indian market lacks the quality uni-strain product of the probiotics as Nutraceuticals and as Health enhancing drug product. Global market is full of multi-strain microbes dosage form. The best mode to utilize Nutraceutical aspect of probiotics is to convert in dry form. These purified colony; later converted into the solid dry form by spray dry (JISL mini-spray drier) technique. Starch, lactose were used as thermo-protective agent while heat drying technique. Anti-microbial screenings carried out along with In vitro cytotoxicity studies. In vitro cytotoxicity screening evaluated that Lactobacillus powder showings anti-cancer activity nearly same as standard drug with no side effects. Dry powder increased the shelf life of the microbes that resulted in maintenance of viability and activity.

Release of toxic and recalcitrant chemicals including synthetic dyes from industries profoundly affects soil fertility and aquatic life. Use of physical and chemical methods for removal of dyes creates disposable problem of remaining dye sludge, whereas biotechnological approach provides viable, less sludge and eco-friendly method. In the present study, a fungus Aspergillus sp. isolated from the soil was employed for decolorization and degradation of five dyes. At first the biodegradation of these dyes was first studied in potato dextrose broth and maximum decolorization of 80.86, 40.05, 92.44, 38.12 and 95 % of decolorization was observed for Methylene blue, Bromophenol blue, Congo red, Malachite green and Rose Bengal respectively. As the maximum decolorization was observed with Congo red and Rose Bengal, the effect of different parameters like carbon source, nitrogen source, pH and temperature was also studied in these dyes. Maximum decolorization of (91 and 52 %) was achieved when supplemented with carbon source, (87 and 46 %) with nitrogen source, (86 and 51 %) at pH at 4–5, (90 and 41 %) at 25–30 °C for Congo red and rose Bengal respectively. The degradation of dyes was observed by the change in original color and visual disappearance of color from the fungus-treated cultures. Degradation/decolorization of dyes was also observed as accumulation of dyes by the fungal mycelium, and it was confirmed by the presence of colored fungal mycelium in fungus-treated cultures. Biosorption studies were also done by using different concentrations of dyes and dead and inactive mycelial mat of Aspergillus sp. This indicated that binding of the dye to dead mycelial mat occurred rapidly with more than 50 % within 1 day and the dye absorbed to the mat increased with increase in the dye concentration, a maximum of 93 and 90 % decolorization was attained at 5 mg Congo red and rose Bengal concentrations with same mycelia mat and maximum of 92 and 88 % decolorization was attained at 750 mg of dead mycelial mat concentration with same dye concentration. Thus the decolorization achieved by metabolically inactive mycelium was equal to that attained by the live mycelium and Aspergillus sp. showed maximum decolorization with Congo red and Rose Bengal dyes. Further this can be commercially used to treat industrial effluents.

Anticancer activity of marine macro algae (seaweeds) Sargassum wightii, Ulva fasciata and Gracillaria corticata against HeLa, K-562 and MDA-MB cell lines was studied. During the present investigation crude extracts of seaweeds were prepared using soxhlet apparatus. Hexane, Butanol and Methanol were used as solvents. Crude extracts were dissolved in DMSO. In vitro anticancer activity of seaweeds at various concentrations (12.5–200 μg/ml) was studied against the chosen cell lines using MTT assay (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, a yellow tetrazole). Butanolic extract of Gracillaria corticata (200 µg/ml) has shown greater anticancer activity than hexane and methanolic extracts of G. corticata and butanol, hexane and methanolic extracts of Sargassum wightii and Ulva fasciata.

Diabetes mellitus is considered to be one of the leading causes of death in adult and children. The most challenging complex heterogeneous group of conditions is prevalent in developing countries as well as the western world is T II DM. Rise in upcoming cases of diabetes has posed it as a global health problem. The predisposition factors are responsible for identifying at risk individuals for this polygenic disorder. Several controllable and non-controllable predisposition factors are responsible for identification of at-risk individuals to T II DM. The complex interactions between genetic and environmental and biochemical predisposition factors unmask the advancement of the disease. This review gives brief description and table form of the work done by the researchers for predisposition of Type II diabetes and some of the related complications among Asian Indian phenotypes and world wide.

The ability to grow plant tissues in vitro and to control their development forms the basis of many practical applications in agriculture, horticulture and industrial chemistry. Establishment of a cell, tissue or organ culture and regeneration of plantlets under in vitro conditions has opened up new avenues in the area of plant biotechnology. Micropropagation is one such method where the plants are grown under in vitro conditions to provide a better survival rate and mass propagation in a short time, it is achieved through the establishment of the explants and their initial growth in vitro, followed by transferring them onto the field or in a greenhouse. This has triggered a great interest in understanding plant physiology and performing a comprehensive study regarding the change in physiology and morphology under influence of various useful bacterial combinations. In this review, plant growth promoting rizobacteria (PGPR) were used in the bio priming of the micro propagated plants (banana), these plants showed increased growth of the root length; shoot length, internode diameter and number of leaves. All parts of the banana plant have medicinal applications: the flowers in bronchitis and dysentery and on ulcers; cooked flowers are given to diabetics; the astringent plant sap in cases of hysteria, epilepsy, leprosy, fevers, hemorrhages, acute dysentery and diarrhea, and it is applied on hemorrhoids, insect and other stings and bites; young leaves are placed as poultices on burns and other skin afflictions; the astringent ashes of the unripe peel and of the leaves are taken in dysentery and diarrhea and used for treating malignant ulcers; the roots are administered in digestive disorders, dysentery and other ailments; banana seed mucilage is given in cases of diarrhea in India. The changes in protein expression as a consequence of environmental stimuli were evaluated which led to the conclusion that the bio hardened plants showed high protein content but had the similar genetic content when compared against the control plants. Thus increase in the protein content enhances these medicinal qualities of banana plant. Further, a fidelity test was conducted on the bio primed plants to check the genetic variations.

Plant growth and developmental processes are very much regulated by certain chemical substances called Growth regulators. Growth regulators are known to improve the physiological efficiency including photosynthetic ability and can enhance the effective partitioning of accumulates from source and sink in the field crops. The present study was conceptualized and executed with the prime objective of study the effect of chlormequat chloride, NAA, Mepiquat chloride and Brassinosteroids on morphological, physiological and biochemical parameters of soybean. The field trial was conducted following randomized block design with nine treatments replicated thrice. The basic material for the present investigation consisted of soybean cv. Js-335 and two growth promoting (NAA and Brassinosteroid) and growth retarding substances (chlormequat chloride and mepiquat chloride). These growth regulators were sprayed at flower initiation stage. The Morphophysiological parameters, namely, Plant height, number of branches, number of trifoliates per plant, dry matter accumulation in leaf, stem and reproductive parts, LAI, CGR and RGR was observed to increase significantly with the application of NAA (20 ppm) and brassinosteroid (25 ppm). However, it decreased with the application of chlormequat chloride and mepiquat chloride. The Biochemical parameters, namely, chlorophyll content was observed to increase significantly with application of NAA (20 ppm), brassinosteroid (25 ppm), mepiquat chloride 5 %, AS (5 %) and chlormequat chloride 50 % SL at different concentrations compared to control and water spray but whereas fluorescence emission and photosynthetic rate were noticed to be non-significant. A significant increase in the seed protein content was also noticed with the application of NAA (20 ppm), brassinosteroid (25 ppm), mepiquat chloride 5 %, AS (5 %) and chlormequat chloride at different concentrations, compared to control and water spray. In conclusion, the study revealed the superiority of NAA (20 ppm) treatment for majority of the morphological, physiological and biochemical parameters at different growth stages, compared to other growth regulator and control treatments studied in the present investigation for Rabi soybean.

The absolute necessity for rational therapy in the face of rampant drug resistance places increasing importance on the accuracy of malaria diagnosis. Giemsa microscopy and Triple test method Rapid Diagnostic Test (RDT) represent the two diagnoses most likely to have the largest impact on malaria control in this present study. These two methods have their characteristic strengths and limitation. These tests were carried out on 156 patients of endemic areas of Gajapati district of Odisha State. These tests were evaluated by two methods, i.e. microscopic slide test and RDT kit method. RDT kits belong to Advantage MAL CARD Malaria pLDH (Plasmodium Lactate Dehydrogenase) antibody Pre-coated (J. Mitra & Co, India) card test, ParaHIT PfHRP2 (Histidine-Rich Protein 2) antibody pre-coated dip stick (Span Diagnostic Ltd, India) and S.D BIOLINE pf/pv capture antigen MSP (Merozoite Surface Protein) pre-coated card (S.D. BIO Standard Diagnostic Pvt. Ltd, India). A little amount of whole blood (5 μl) was taken using the plastic loop given with the kit, at the same time two slides are also taken. Out of 156 patients it was detected that 128 were negative and 28 were positive according to microscopic detection. Out of these 28 positives 23 were only P. falciparum, 2 were only P. vivax, 3 were having both PF and PV infection detected. On the other hand 32 were positive and 4 were false positive as shown by Advantage MAL CARD and ParaHIT, where as 49 were positive and 21 were false positive by the S.D. MSP pre-coated antigen card test kits.

Brain Derived neurotrophic factor (BDNF) is very well reported in development of neurons and it plays major role in memory and interpretation. It is evident that BDNF is involved in maintaining the equilibrium of body weight and glucose homeostasis mechanism. Out of 96 Subjects included in this study Plasma BDNF level was found low in the patients with Type 2 Diabetes. We didn’t find positive indications wrt association of BDNF G196A (Val66Met) polymorphism in diabetes or obesity. Type 2 Diabetic patients with the complaint of Joint pains were found to have even lower Plasma BDNF levels as compared to the diabetic patients without any Neurological problems or joint pains. Worsening BDNF Levels may be an alarming factor for type 2 diabetic patients with respect to development of Neurological disorders.