Bio Technology

The study aimed at detoxifying JCSM using biological treatments (spontaneously, fungi cocktail and specific). Shimadzu HPLC/ UV was used to quantify phorbol esters in Jatropha curcas seed meal under isocratic conditions. After six days of fermentation, cocktail of fungi, and spontaneous fermentation of JCSM was effective in reducing PE concentration. Defattening seed meal using cold maceration in petroleum ether reduced phorbol-12-myristate-13-acetate (PMA) in the JCSM from 40.42 to 10.9 ng/ml prior to solid state fermentation. Spontaneous fermentation of defatted JCSM further reduced PMA from 10.9 to 5.05 ng/ml (53.6%). Fungi isolated during spontaneous fermentation included, Aspergillus niger (B), Aspergillus flavus (A), Rhizopus Stolonifer (C), Penicillium chrysogenum (D) and Fusarium species. The reduction of PE ranged between 10.71 – 1.46 ng/ml when single or a combination of the fungi isolates were used in solid state fermentation of the defatted seed meal. P. chrysogenum reduced phorbol ester levels (PE) in defatted seed 76.5%, R. stolonifer 43.3%, A. flavus 35.6% and A. niger 1.72%. When fungi isolates were paired, A. niger + P. chrysogenum (BD) reduced PE levels by 13.7%, A. flavus + A. niger 64.5%, A. flavus + P. chrysogenum 70.3%, A. flavus + R. stolonifer 78.8%, A. niger + R. stolonifer 86.6% and R. stolonifer + P. chrysogenum 86.6%. The treatment combination of A. niger + R. stolonifer + P. chrysogenum reduced PE levels by 84.0%, A. flavus + A. niger + R. stolonifer to 67.2% and A. flavus + R. stolonifer + P. chrysogenum to 62.3%. P. chrysogenum was effective as a single isolate but its activity was suppressed when paired with A. niger. Treatments of the deffated seed with, A. flavus + R. stolonifer; A. niger + R. stolonifer; R. stolonifer + P. chrysogenum; and A. niger + R. stolonifer + P. chrysogenum were the only combination that exceeded the phorbol reductive activity of P. chrysogenum in. It could therefore be concluded that Jatropha curcas seed meals can be detoxified by solid state fermentation.

Intense interest in Asparaginase has resulted from the discovery of its ability to inhibit growth of tumors in mouse, rat and dog to suppress human leukamias in clinical trails. L-Asparaginase is an effective antineoplastic agent, used in the acute lymphoblastic leukemia.Asparaginase catalyzes the deamination of asparaginases into L-aspartic acid and ammonia. The aim of the present investigation was to study production of asparaginase from agricultural waste like citrus limetta pulp using solid state fermentation (SSF).Citrus limetta pulp used as a sole source for growth in SSF showed maximum enzymes production. Optimized process parameters like incubation time: 72 hrs; incubation temperature:280C; pH of the culture medium: 7.5; and moisture content: 60% v/w gave an overall yield of 101 U/g.

The comparative genome analysis of Streptococcus pyogenes and Streptococcus equi sps equi 4047 depict exon, Untranslated Region (UTR), conserved noncoding sequences (CNS), contigs, mobile genetic elements, Insertion sequence (IS) elements, transposons, plasmids and shows high degree of heterogeneity evidenced by the large number of single nucleotide polymorphisms (SNPs). Thus UTR, CNS, and IS elements can be used as drug target.

This study was conducted to explore the biofilm forming capabilities of extended spectrum beta-lactamase (ESBL) producing and non-producing Klebsiella pneumoniae strains by three different methods and to compare the methods as well. Two ESBL producers and two non-producers strains were subjected to biofilm detection methods. These isolates were previously confirmed by standard methods and API (Analytical Profile Index) scoring. Their ESBL production capabilities were confirmed by double discs synergy test as recommended by the Clinical and Laboratory Standards Institute. Biofilm formation was detected by Congo Red Agar method (CRA), Test Tube method (TTM) and Microtiter Plate Assay method (MTP). Both the ESBL producing Klebsiella pneumoniae were characterized as strong biofilm formers. Between the two ESBL non-producers, one was assessed as weak and the other as moderate biofilm former by quantitative TTM and MTP. The MTP and quantitative TTM were considered superior to qualitative TTM and CRA method. Our findings highlighted the relatively higher biofilm forming ability by the ESBL positive Klebsiella pneumoniae that may additionally contribute to their resistance against extended spectrum antibiotics. We can conclude that K. pneumoniae strains, isolated from blood, to form biofilm have a significant association with ESBL production.

With increasing demand for steel and depletion of high-grade iron ore deposits, more research efforts are being directed toward extending the life of existing ore reserves and developing technology to treat low-grade iron ore resources. Interaction between Bacillus subtilis and iron ore minerals such iron oxide and, quartz brought about significant surface chemical changes. Quartz was rendered more hydrophilic, while iron oxide became more hydrophobic mineral after bacterial interaction. The surface properties were studied using zeta potential, SEM, adhesion of strains to the minerals’ surface and contact angle. In this work Bacillus subtilis was isolated from Egyptian iron ore surface. The results revealed that a strong interaction was occurred. A concentrate contains 90.88% Fe2O3 [64 % Fe (total)] and 4.46% SiO2 was obtained through bio-flotation from a feed contains 70.04% Fe2O3 [49.33% Fe (total)] and 21.18% SiO2.

The primary objective was to evaluate if batch staining of malaria blood films results in false positive smears. False positive smears (>1%) may cause a serious underestimate of a drug’s or vaccine’s protective efficacy, as well as affect evaluation of diagnostics, estimates of malaria prevalence, and clinical management. Thick blood films may float from a glass slide during staining and adhere to other films if batch staining is used resulting in false positive readings. Venous blood in EDTA anticoagulant from malaria positive samples of ? 20 parasites per high power field and a true negative sample was utilized to make thick and thin smears. Two true negative smears were stained with Giemsa stain with eight positive smears in batch in Coplin jars for 10 minutes or overnight. Two control negatives were stained alone with the same batch of stain. Blinded microscopists read these slides using a rereading paradigm. Thick film loss was graded by gross appearance ranging from 0 (none) to 4+ (> ¾ loss). A total of 602 slides were evaluated in this study, of which 392 were true positives (65%) and 210 (35%) were true negatives. Of the true negatives, 110 were batch stained with true positives, and 100 were true negative controls stained alone. Of the initial readings, 11-20% were reported falsely positive. “Fishing” or cross-contamination was infrequently noted by one of the microscopists, but was uniformly present in these smears on reexamination. Of the true positive smears (high density), 1-3% were read falsely negative. On reexamination of these slides, the cause was found to be reporting of results from very poor quality smears. Thick film loss was clearly more severe for the positive slides with 10 minute versus overnight drying (means score 0.97 vs 1.97, p <0.001). This experiment confirmed that false positive smears result from cross-contamination during batch staining using methods employed today. Since low frequencies of false positive smears can adversely impact research and product development results, single slide staining should become the norm in this setting. Reporting of false negative results occurred in malaria smears with high densities of parasites. Microscopists should be trained not to report results when smear quality is not adequate.

Visceral leishmaniasis (VL) is an important cause of morbidity and mortality that is characterized by fever, lymphadenopathy and hepato-splenomegaly. Hepatic toxicity greatly contributes to VL morbidity. This study aimed to evaluate liver damage in Sudanese patients with VL as evidenced by apoptosis and lipid metabolism derangement. In a prospective analytical, hospital-based and case-controlled study and following informed consent, eighty patients with parasitologically confirmed visceral leishmaniasis and eighty apparently healthy age and sex unmatched volunteers [comparators] were enrolled in the study. Serum cytochrome C was measured by ELISA while serum lipids were measured using BioSystems A15 Chemistry Auto-analyzer. Cytochrome C concentrations in VL patients were significantly higher compared to apparently healthy volunteers with no significant difference between pre and post-treatment samples. Patients with VL showed marked hypo-cholestereamia, very low serum levels of LDL and HDL with most patients showing markedly increased triglycerides levels. Deranged lipid metabolism in VL patients could be due to hepatotoxicity or sequestration and/or degradation of lipoproteins in enlarged livers and spleens. In conclusion, hypocholestereamia, low levels of LDL/HDL, high triglycerides levels and increased serum cytochrome C are important features of hepatotoxicity in VL. Increased serum cytochrome C level is probably an important surrogate marker of hepatocytes apoptosis.

Elaeocarpus ganitrus Roxb. (Elaeocarpaceae) is a large evergreen broad-leaved tree and found in Himalayan range in India and Nepal. Fruits and leaves are known for various medicinal properties and used in traditional medication system for the treatment of diseases. In this study, antimicrobial activity of the aqueous extract of leaves of E. ganitrus was tested against clinical isolates of bacteria and fungi. In vitro antimicrobial activity was performed by agar well diffusion method on Mueller Hinton agar and Sabouraud Dextrose agar for bacterial and fungal cultures respectively. The extract exhibited a broad spectrum of antimicrobial activity as it inhibited the growth of Staphylococcus aureus, Bacillus cereus, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Penicillium sp, Aspergillus flavus, Candida albicans and C. tropicalis. The extract showed maximum relative percentage inhibition against B. cereus (124.16%). Minimum inhibitory concentration test was performed by modified agar well diffusion method. Minimum inhibitory concentration values of the extract varied from 125-2000 µg/ml; however minimum value was reported against B. cereus and A. flavus (125 µg/ml). The results indicate the potential use of E. ganitrus leaves for the development of antimicrobial compounds.

In this study, two cotton cultivars viz. LD 694 (Gossypium arboreum L.) and LH 2076 (Gossypium hirsutum L.) were used to investigate factors influencing efficiency of transformation from the shoot apices of aseptically germinated seedlings for the Agrobacterium mediated transformation using the Agrobacterium strain GV3101 having the plasmid pPZP200 vector, carrying cry1Ac gene. The objective of study was to develop genotype independent Agrobacterium mediated transformation method. Antibiotics sensitivity assay showed that Kanamycin @ 50mg/l inhibited the regeneration of shoot tips in both LD 694 and LH 2076 genotypes and Cefotaxime @ 250mg/l check the bacterial growth. For Agrobacterium mediated transformation, a dilution of 500 µl /25ml of water using dip method and co-cultivation for 72 hours were resulted in better infection and survival of agroinfected explants with GV3101 strain in both genotypes. Average survival of 11.42% and 13.51% was recorded for Agrobacterium transformed shoots on selection medium supplemented with 50 mg/l Kanamycin and 250mg/l Cefotaxime in LD 694 and LH 2076 genotypes, respectively. GUS expression of transformed tissues ranged from 5.5-12.0 % in shoot tips. PCR analysis confirmed the presence of Cry 1Ac gene in putative transformed tissues.

Inhibition of ?-amylase, enzyme that plays a role in digestion of starch and glycogen, is considered a strategy for the treatment of disorders in carbohydrate uptake, such as diabetes and obesity, etc. Plants are an important source of chemical constituents with potential for inhibition of ?-amylase and can be used as therapeutic or functional food sources. In this study methanolic extracts from Mimosa pudica plant source that have been tested for ?-amylase & acid phosphatase inhibitory activity has been done. Amongst the phyto-constituents that have been investigated, flavonoids are one of them that demonstrated the highest inhibitory activities with the potential of inhibition related to number of hydroxyl groups in the molecule of the compound.The inhibition of acid phosphatase could be due to the rupture of lysosomal membrane in the presence of Mimosa pudica. Methanol crude extracts of the (whole plant) of Mimosa pudica (Mimosaceae) were screened in vitro for cytotoxicity studies by brine shrimp lethality bioassay and antioxidant activity using the 1,1-diphenyl-2-picrylhydrazyl-hydrate (DPPH) free radical scavenging assay. The methanol crude extracts of the whole plant showed potential cytotoxic activities (IC50 82.50?g/ml). The methanol crude extract of the whole plant showed moderate antioxidant activity (IC50 48.54?g/ml) compared to Rutin (IC50 34.60?g/ml). The % of inhibition of alpha Amylase & Acid Phosphatase of Mimosa pudica was found to be 41% & 47.8% in the concentration of 5mg/ml respectively with membrane stability 61.5%. The overall experimental results suggested that biologically active constituents present in the methanolic extract of Mimosa pudica and justifies its use in folkloric remedies.