In vitro callus induction and estimation of plumbagin content from Plumbago auriculata Lam.

The medicinal plant Plumbago contains a very potent secondary metabolite, plumbagin having many therapeutic properties. Callus culture was induced using explants, leaf, stem and shoot apex, from P. auriculata. Murashige and Skoog media fortified with various growth hormones like NAA, IAA, IBA and 2, 4-D individually and in various combinations were checked for callus induction. Among the growth hormones used, 1 mg/L 2, 4-D showed best callusing. The hormonal combinations of 1 mg/L IAA and 1.5 mg/L NAA in the media exhibited best callus induction using stem internode as an explant. Plumbagin content from root, stem, leaf and callus was analyzed by using thin layer chromatographic technique. The callus derived from stem showed comparable plumbagin content to the in vivo plant parts. Quantitative spectrophotometric analysis of plumbagin from plant samples and callus indicated that plumbagin content was maximum in roots which was followed by callus, stem and leaf samples respectively. Generation of in vitro sources for plumbagin, for therapeutic applications will serve as a continuous supply and will contribute to preserve the natural plant recourses.

A robust HIV-1 viral load detection assay optimized for Indian sub type C specific strains and resource limiting setting

BACKGROUND: Human Immunodeficiency Virus Type 1 (HIV-1) viral load testing at regular intervals is an integral component of disease management in Acquired Immunodeficiency Syndrome (AIDS) patients. The need in countries like India is therefore an assay that is not only economical but efficient and highly specific for HIV-1 sub type C virus. This study reports a SYBR Green-based HIV-1 real time PCR assay for viral load testing and is designed for enhanced specificity towards HIV-1 sub type C viruses prevalent in India. RESULTS: Linear regression of the observed and reference concentration of standards used in this study generated a correlation coefficient of 0.998 (p<0.001). Lower limit of detection of the test protocol was 50 copies/ml of plasma. The assay demonstrated 100% specificity when tested with negative control sera. The Spearman coefficient of the reported assay with an US-FDA approved, Taqman probe-based commercial kit was found to be 0.997. No significant difference in viral load was detected when the SYBR Green based assay was used to test infected plasma stored at -20°C and room temperature for 7 days respectively (Wilcoxon signed rank test, p=0.105). In a comparative study on 90 pretested HIV-1 positive samples with viral loads ranging from 5,000 - 25,000 HIV-1 RNA copies/ml and between two commercial assays it was found that the later failed to amplify in 13.33% and 10% samples respectively while in 7.77% and 4.44% samples the copy number values were reduced by >0.5 log value, a figure that is considered clinically significant by physicians. CONCLUSION: The HIV-1 viral load assay reported in this study was found to be robust, reliable, economical and effective in resource limited settings such as those existing in India. PCR probes specially designed from HIV-1 Subtype C-specific nucleotide sequences originating from India imparted specificity towards such isolates and demonstrated superior results when compared to two similar commercial assays widely used in India.

Characterization of Dairy Effluents by Physicochemical Parameters.

Aims: The dairy industry faces growing scrutiny of its environmental stewardship. The potential impact of an individual operation on the environment varies with animal concentration, weather, terrain, soils, and numerous other conditions. It is hoped that management practices found on dairy industry will benefit by the management practices. So properly applied the Management Practices, the factual study of dairy effluent by various physico-chemical characters concern for environmental health and safety. Study Design: Effluent samples were collected from dairy industries. The samples were characterized by physical parameters like pH, temperature, TS, etc. and chemical parameters BOD, COD, DO etc. Place and Duration of Study: Effluent samples were collected from dairy industry of district Kolhapur Maharashtra (India).Physico-chemical characteristics of the effluent during the months between March to August 2011. Methodology: Total 4 samples of dairy effluents were collected by composite sampling at the time 9, 12, 3, 6 o’clock per day and stored at 4oc for further analysis. Then on the next day in quintet it was subjected to analyze the physicochemical parameters like Temperature, pH, DO, TDS, TSS, TS, BOD, COD, chloride, Sulphate, oil and Grease. Results: The study revealed that the dairy effluent is slightly alkaline in nature, and high temperature, BOD & COD values obtained by the analysis of dairy effluents indicate the presence of heavy load of organic substances. Also a higher temperature and oils and Greases which lower the dissolved oxygen activities can cause serious problems in disposal of waste water. Above the standard value suspended and dissolved organic solids are responsible for creating nuisance. Conclusion: Dairy industry tested in this study was found high levels of pH, BOD, COD, TSS It is very important that proper waste water treatment systems should be installed for the protection of the environmental health and for the ecological balance.

Isolation of Microorganisms from Dairy Effluent.

Aim: Every Dairy industry has problems of effluent treatment. This can be revealed by effective treatment of the effluent. The effective treatment can be done by using microorganisms to stabilize the organic and inorganic load of the effluent. The aim of the present work is to study the dairy wastewater micro biota and to identify some new active strains which can bring about fast biodegradation of the organic compounds. Study Design: Isolation and determination of bacteriological characteristics of the dairy effluents. Methodology: Studies were carried out to isolate the microorganisms from collected effluent sample from the dairies under studies. Isolation of microorganisms was done by primary screening, Cultural Characterization, Biochemical characterization and Identified by using Bergey’s Manuals of Systematic Bacteriology. Results: During 2011-2013 from two different districts of Maharashtra (India), dairy industry effluents were collected for the isolation of micro organisms. Effluent samples were collected as per Jacksch and piper method and primary screening was done and totally 7 Isolates were screened out. These isolates were characterized on nutrient agar at room temperature for 24 hrs. Isolates were observed for the cultural characters like size, shape, colour, margin, elevation, opacity and consistency and morphological characters like Gram nature, sporulation, shape and arrangement of cells, motility etc. and were recorded. Physio-biochemical characterization was followed by biochemical tests for enzymatic activities like catalase, oxidase, nitrate reduction, urease, caseinase etc and carbohydrates utilization tests for lactose, maltose, inositol, xylose etc. performed to check their ability for metabolization. On the basis of these characteristics, isolates were identified by using Bergey’s Manuals of Systematic Bacteriology. The identified Bacterial Isolates were of Genus Lactobacillus, Bacillus, Staphylococcus, Enterococcus, Listeria etc. Conclusion: These Bacterial isolates have the ability to utilize the components like nitrate, starch, gelatin, sugars like sucrose, maltose, lactose etc. which was confirmed by the biochemical tests. Bacterial flora from the effluents can be identified and efficiently applied for the biological treatment of the dairy effluents.