Assessment of Iron Biosorption Potential by Live and Dead Biomass of Bacillus subtilis (MN093305) from Aqueous Solution.

Heavy metals polluted aquatic ecosystems and become a global environmental issue due to their toxic effect on all forms of ecosystems and further on all forms of life. Heavy metals are non- degradable and accumulated in different life forms by accumulating in the food chain; this increases the need for the development of a sustainable method for the removal of these metals. Biosorption is an eco-friendly and cost-effective convenient technique of heavy metal bioremediation from the contaminated aquatic ecosystem. The current investigation involves biosorption of iron using Bacillus subtilis strain (MN093305) isolated from Ganga river at different physical parameters with the highest rate of biosorption was 96.64%, 98.91%, 97.88%, and 99.44% at pH 5, 60 min incubation period, 35 °C temperature and 2.5 mg/ml of biomass respectively for dead biomass. Living biomass biosorption rate was 87.32%, 96.74%, 96.94% and 95.02% at pH 7, 72 h, 35 °C and 2.5 mg/ml respectively. Functional groups involved in the biosorption of iron by Bacillus subtilis were fitted to a second-order kinetic model. Langmuir and Freundlich's isotherm are used to evaluate data; both isotherms indicate iron absorption as a favorable process.

Production of polyhydroxyalkanoates from renewable resources: a review on prospects, challenges and applications.

Bioplastics replace synthetic plastics of petrochemical origin, which contributes challenge to both polymer quality and economics. Novel polyhydroxyalkanoates (PHA)-composite materials, with desirable product quality, could be developed, thus targeting the global plastics market, in the coming years. It is possible that PHA can be a greener substitute for their petroleum-based competitors since they are simply decomposed, which may lessen the pressure on municipal and industrial waste management systems. PHA production has proven to be the bottleneck in industrial application and commercialization because of the high price of carbon substrates and downstream processes required to achieve reliability. Bacterial PHA production by these municipal and industrial wastes, which act as a cheap, renewable carbon substrate, eliminates waste management hassles and acts as an efficient substitute for synthetic plastics. In the present review, challenges and opportunities related to the commercialization of polyhydroxyalkanoates are discussed and presented. Moreover, it discusses critical steps of their production process, feedstock evaluation, optimization strategies, and downstream processes. This information may provide us the complete utilization of bacterial PHA during possible applications in packaging, nutrition, medicine, and pharmaceuticals.

Optimization of Growth Conditions to Produce Sustainable Polyhydroxyalkanoate Bioplastic by Pseudomonas aeruginosa EO1.

Polyhydroxyalkanoates (PHAs) are intracellularly synthesized by bacteria as carbonosomes that exhibit biodegradable thermoplastics and elastomeric properties. The use of cheaper edible oils as a source of carbon assists in the reduction of the production cost of such biopolyesters. In this work, different edible oils, such as groundnut oil (GNO), mustard oil, sesame oil, and soybean oil (SBO) were used to check their effect on PHA production from Pseudomonas aeruginosa EO1 (MK049902). Pseudomonas aeruginosa EO1 was used in a two-stage production system. In the first stage, bacterial growth was favored and, in the second, PHA was synthesized. GNO was found as the best carbon source for PHA production. The use of 2% (v/v) GNO, rich in saturated fatty acids, allowed PHA content of 58.41% and dry cell weight (DCW) of 10.5g/L at pH7 and temperature 35°C for 72h. Groundnut has a high potential for oil production and for the diversification of co-products with some potential of value aggregation. Such a perennial and sustainable species will almost certainly meet the criteria for becoming a significant commercial oilseed crop. Fourier transform infrared spectroscopy (FTIR) spectra showed strong characteristic bands at 1,282, 1,725, 2,935, 2,999, and 3,137cm-1 for the PHA polymer. Gas chromatography-mass spectrometry (GC-MS) detects the presence of PHA copolymers.

Head growth of undernourished children in rural Nepal: association with demographics, health and diet.

BACKGROUND: Brain development in early childhood is a key determinant of later cognition, social achievement and educational success. Head circumference (HC) measurements are a simple method to assess brain growth, yet reports of these measurements are uncommon in nutritional surveys of undernourished children. OBJECTIVE: To evaluate HC measurements in a population of rural Nepali children and relate these measurements to demographics, health and diet. METHODS: An observational study of head growth was nested within a longitudinal evaluation of a livestock-based agricultural intervention in rural Nepal. Between 538 and 689 children (aged 6 months to 8 years) were measured (height, weight, HC) at each of six survey visits. A total of 3652 HC measurements were obtained. Results were converted to Z-scores (WHO Anthro). RESULTS: Mean head circumference Z-scores (HCZ) diminished progressively over the first 4 years of life; a decline of 30% occurred between 3 and 4 years of age (-1.73 to -2.45, P < 0.0001). Overall, 56% of HCZ were <-2. Gender-adjusted HCZ (but not other measurements) were significantly lower for girls than boys [mean (SD) -2.31 (1.0) vs -1.99 (0.094), P < 0.0001]; girls more often had microcephaly (61% vs 50%, P < 0.0001). For children <3 years of age, HCZ were better in those who had eaten two or more animal-source foods (ASFs) within the previous 24 h [-1.69 (.05) vs -2.08 (0.10), P = 0.001] than in those who had eaten none or only one; HCZ correlated with the number of ASFs consumed (P < 0.001). Regression analyses demonstrated that the main determinants of HCZ were age, weight-for-age Z-scores (WAZ) and gender; 43% of the variance in HCZ in younger children was explained by WAZ and ASF consumption. CONCLUSION: HCs reflect brain size in young children; brain size is linked to cognitive function. Poor head growth represents another facet of the 'silent emergency' of child undernutrition. Routine HCZ assessments may contribute to better understanding of the links between poverty and cognitive development.

Community development and livestock promotion in rural Nepal: effects on child growth and health.

BACKGROUND: More than 50% of children in Nepal are malnourished. Economic growth and poverty reduction are not always sufficient to improve the health and nutritional status of children. Heifer Nepal uses livestock training as a tool for community development and poverty alleviation but does not directly address child health and nutrition. OBJECTIVE: To systematically assess the effects of Heifer activities on child health and nutrition. METHODS: The study was a 2-year, longitudinal, randomized, controlled trial in six communities in Nepal (both Terai and hills), pair-matched for specific characteristics, randomly assigned to receive Heifer community development activities at baseline (intervention) or 1 year (control). At 6-month intervals over a period of 2 years, child anthropometric and comprehensive household surveys were performed. RESULTS: Four hundred fifteen households were enrolled containing 607 children 6 months to 5 years of age. The intervention and control communities were equivalent for baseline socioeconomic status, household size, ownership of land and animals, and child nutrition and health. At 12 months (prior to animal donations), the Terai intervention group had improved child weight (p = .04), improved child height (p = .05), and reduced sick days (p = .03), as well as increased household income (p = .004), increased ownership of animals (p = .04) and land (p = .04), and improved sanitation practices (p < .01). In all districts, longer participation in Heifer activities corresponded to more improvement in child height-for-age z-scores. CONCLUSIONS: Heifer interventions resulted in improved socioeconomic status and household income per family member. Children under 60 months of age in the intervention group had greater incremental improvement in height-for-age and weight-for-age z-scores than children in the control group, and longer participation in Heifer activities was associated with better growth. Poverty alleviation programs, such as Heifer, may indirectly benefit child growth.

Small-molecule quinolinol inhibitor identified provides protection against BoNT/A in mice.

Botulinum neurotoxins (BoNTs), etiological agents of the life threatening neuroparalytic disease botulism, are the most toxic substances currently known. The potential for the use as bioweapon makes the development of small-molecule inhibitor against these deadly toxins is a top priority. Currently, there are no approved pharmacological treatments for BoNT intoxication. Although an effective vaccine/immunotherapy is available for immuno-prophylaxis but this cannot reverse the effects of toxin inside neurons. A small-molecule pharmacological intervention, especially one that would be effective against the light chain protease, would be highly desirable. Similarity search was carried out from ChemBridge and NSC libraries to the hit (7-(phenyl(8-quinolinylamino)methyl)-8-quinolinol; NSC 84096) to mine its analogs. Several hits obtained were screened for in silico inhibition using AutoDock 4.1 and 19 new molecules selected based on binding energy and Ki. Among these, eleven quinolinol derivatives potently inhibited in vitro endopeptidase activity of botulinum neurotoxin type A light chain (rBoNT/A-LC) on synaptosomes isolated from rat brain which simulate the in vivo system. Five of these inhibitor molecules exhibited IC(50) values ranging from 3.0 nM to 10.0 µM. NSC 84087 is the most potent inhibitor reported so far, found to be a promising lead for therapeutic development, as it exhibits no toxicity, and is able to protect animals from pre and post challenge of botulinum neurotoxin type A (BoNT/A).

Prevention of aggregation and autocatalysis for sustaining biological activity of recombinant BoNT/A-LC upon long-term storage.

Protein aggregation during expression, purification, storage, or transfer into requisite assay buffers hampers the use of proteins for in vitro studies. The formation of these aggregates represents a major obstacle in the study of biological activity and also restricts the spectrum of protein products being available for the biomedical applications. The catalytic light chain of botulinum neurotoxin type A undergoes autocatalysis and aggregation after purification upon long-term storage and freeze-thawing. In present study the conditions for the high level expression and purification of biologically active light chain protein of botulinum neurotoxin were optimized from a synthetic gene. Several co-solvents were screened in order to prevent autocatalysis and aggregation of rBoNT/A-LC. The effect of the co-solvents is studied on endopeptidase activity during long term storage of the recombinant protein. The purified rBoNT/A-LC was also evaluated for its immunogenicity.

Potentiation of antimicrobial activity of ciprofloxacin by Pelargonium graveolens essential oil against selected uropathogens.

The recent approach of using herbs and antibiotics in combination constitutes a strategy to overcome the problems of resistance and side effects associated with conventional antibiotics. In the present study, the antimicrobial effect of Pelargonium graveolens L' Hér essential oil in combination with ciprofloxacin was evaluated on uropathogens, namely, Klebsiella pneumoniae KT2, Proteus mirabilis PRT3 and Staphylococcus aureus ST2. Minimum inhibitory concentrations of P. graveolens essential oil and ciprofloxacin were determined by the microbroth dilution method and further, the interaction between these two agents was studied by a checkerboard method. The fractional inhibitory concentration index (FICI) was calculated to be 0.375 for both K. pneumoniae KT2 and P. mirabilis PRT3, while for S. aureus ST2 it was found to be 0.5. The values of FICI for the tested microorganisms were found to be ≤0.5, which indicates synergism between P. graveolens essential oil and ciprofloxacin. The concave shaped curve in the isobolograms also depicted a synergistic effect of P. graveolens essential oil and ciprofloxacin against the tested microorganisms. Hence, the synergistic action of P. graveolens essential oil and ciprofloxacin may be applied for the treatment of UTIs, which have hitherto been treated by using only synthetic drugs.

Characterization of LC-HCC fusion protein of botulinum neurotoxin type A.

Botulinum neurotoxins (BoNTs) are highly potent toxins that inhibit neurotransmitter release from peripheral cholinergic synapses. The gene for encoding the full length light chain with H(CC) (binding) domain of Clostridium botulinum neurotoxin A was synthesized and cloned into a bacterial expression vector pQE30-UA and produced as an N-terminally six-histidine-tagged fusion protein (rBoNT/A LC-H(CC)). This protein was expressed in two different strains of Escherichia coli namely BL21(DE3) and SG13009. Expression at 37 °C revealed localization of rBoNT/A LC- H(CC) in inclusion body whereas it was expressed in soluble form at 21°C. The recombinant fusion protein was purified by nickel affinity gel column chromatography and identified by monoclonal antibody and peptide mass fingerprinting. The recombinant protein was shown to bind with synaptic vesicles and gangliosides (GT1b) using enzyme-linked immunosorbent assay. The rBoNT/A LC-H(CC) was also found to be highly active on its substrate (SNAP-25) from rat brain, indicating that the expressed and purified rBoNT/A LC-H(CC) protein retains a functionally active conformation. Biologically active recombinant fusion protein was also evaluated for its immunological potential.

Botulinum toxin: bioweapon & magic drug.

Botulinum neurotoxins, causative agents of botulism in humans, are produced by Clostridium botulinum, an anaerobic spore-former Gram positive bacillus. Botulinum neurotoxin poses a major bioweapon threat because of its extreme potency and lethality; its ease of production, transport, and misuse; and the need for prolonged intensive care among affected persons. A single gram of crystalline toxin, evenly dispersed and inhaled, can kill more than one million people. The basis of the phenomenal potency of botulinum toxin is enzymatic; the toxin is a zinc proteinase that cleaves neuronal vesicle associated proteins responsible for acetylcholine release into the neuromuscular junction. As a military or terrorist weapon, botulinum toxin could be disseminated via aerosol or by contamination of water or food supplies, causing widespread casualties. A fascinating aspect of botulinum toxin research in recent years has been development of the most potent toxin into a molecule of significant therapeutic utility . It is the first biological toxin which is licensed for treatment of human diseases. In the late 1980s, Canada approved use of the toxin to treat strabismus, in 2001 in the removal of facial wrinkles and in 2002, the FDA in the United States followed suit. The present review focuses on both warfare potential and medical uses of botulinum neurotoxin.

Biomethanation under psychrophilic conditions.

The biomethanation of organic matter represents a long-standing, well-established technology. Although at mesophilic and thermophilic temperatures the process is well understood, current knowledge on psychrophilic biomethanation is somewhat scarce. Methanogenesis is particularly sensitive to temperature, which not only affects the activity and structure of the microbial community, but also results in a change in the degradation pathway of organic matter. There is evidence of psychrophilic methanogenesis in natural environments, and a number of methanogenic archaea have been isolated with optimum growth temperatures of 15-25 °C. At psychrophilic temperatures, large amounts of heat are needed to operate reactors, thus resulting in a marginal or negative overall energy yield. Biomethanation at ambient temperature can alleviate this requirement, but for stable biogas production, a microbial consortium adapted to low temperatures or a psychrophilic consortium is required. Single-step or two-step high rate anaerobic reactors [expanded granular sludge bed (EGSB) and up flow anaerobic sludge bed (UASB)] have been used for the treatment of low strength wastewater. Simplified versions of these reactors, such as anaerobic sequencing batch reactors (ASBR) and anaerobic migrating blanket reactor (AMBR) have also been developed with the aim of reducing volume and cost. This technology has been further simplified and extended for the disposal of night soil in high altitude, low temperature areas of the Himalayas, where the hilly terrain, non-availability of conventional energy, harsh climate and space constraints limit the application of complicated reactors. Biomethanation at psychrophilic temperatures and the contribution made to night-soil degradation in the Himalayas are reviewed in this article.

Antibodies against recombinant shiga toxin subunit B neutralize shiga toxin toxicity in HeLa cells.

Shigella dysenteriae type 1 and Escherichia coli O157:H7 produce Shiga toxin 1 (Stx) and Shiga toxin1 (Stx1), respectively and these two toxins are almost identical. E. coli O157:H7 is the major cause of diarrhea-associated hemolytic uremic syndrome. Stx and Stx1 are AB5 type of toxin with a molecular weight of 70 kDa, comprising an enzymaticaly-active A subunit (32 kDa) and five receptor-binding B subunits (7.7 kDa). In this study DNA fragment (289 bp, Gene Bank Accn No. EF685161) coding for B chain of Stx was amplified from S. dysenteriae type1 and cloned. Shiga toxin-binding subunit was expressed and purified in native conditions by affinity and gel permeation chromatography with the yield of 5.1 mg/L in shake flask culture. For the purpose of immunization, the polypeptide was polymerized with glutaraldehyde. Hyper immune serum produced in mice reacted with the purified polypeptide and intact Shiga toxin. The anti-StxB antiserum effectively neutralized the cytotoxicity of Shiga toxin towards HeLa cells.

Isolation of hydrocarbon degrading bacteria from soils contaminated with crude oil spills.

A feasibility study was conducted to evaluate the capability of bacterial strains to degrade crude oil under in vitro conditions. Pseudomonas strain PS-I could degrade alkanes (70.69%) and aromatics (45.37%). Alkanes and aromatic fractions separated by column chromatography were analyzed by gas chromatography. In case of Pseudomonas strain PS-I, nC17/Pr, nC18/Ph ratios decreased from 2.5100 to 0.1232 and from 7.2886 to 0.3853, respectively. It was concluded that out of the isolated strains, Pseudomonas strain PS-I, PS-II and PS-III were comparatively better and potent hydrocarbon degraders. Pseudomonas strain PS-I was almost comparable with standard strain of Acinetobacter calcoaceticus in crude oil biodegradation potency.