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Asian Journal of Biotechnology

This study is aimed at investigating the most efficient and cost effective sterilization condition for initiation of sweet potato culture in-vitro. Different concentrations of common or local household bleach (Power zone and Dettol) and non-readily available mercuric chloride as well as variation in their duration for sterilization were used to sterilize sweet potato buds to study their effectiveness in preventing microbial growth in sweet potato culture. The most common microbial contaminants observed in the sweet potato culture were fungi (Fusarium sp., Aspergillus nger, Penicillium sp. and Neurospora crassa). Mericuric chloride in combination with other treatment was found to be the most effective in suppressing microbial contaminants. Dettol in combination with other treatments was found to be ineffective in suppressing the microbial contamination. Duration of 20 min treatment was found to be the most effective in suppressing the microbial contamination but was very toxic to the explants. Treating explants for 10 min (control) was found to be ineffective in suppressing the microbial contamination but non-toxic to explants. Subjecting the sweet potato explants to sterilization for 12 and 15 min were found to be the most efficient, since there was no significant difference between them. These findings will help alleviate much of the burden associated with initiation of sweet potato cultures thereby reducing cost of plantlets production. 2014/07/26 - 11:07

Agrobacterium-mediated transient gene transfer or agroinfiltration is a method to induce transient expression of heterologous proteins in plants. It offers speed, low cost and convenience when compared to traditional plant transformation. This study was aimed to set up a simple but feasible laboratory-scale vacuum agroinfiltration system. Basically, the procedures involved whole plant immersion into transformed Agrobacterium tumefaciens suspension and pressure application via vacuum drawing the agrobacteria into the plant tissues homogeneously. Optimizations of its parameters such as vacuum pressure and treatment duration towards the expression activity of a reporter, Green Fluorescence Protein (GFP) were conducted. The transformed A. tumefaciens strain LBA4404 carrying the recombinant plant viral vector, pEAQ-HT-GFP was infiltrated into the leaves of N. benthamiana via vacuum mode. In general, vacuum pressure and duration of infiltration had significant effects (p<0.05) on transient expression of GFP. Both fluorescence microscopy and Western blotting showed that vacuum agroinfiltration at 600 mmHg for 3 min had the highest and 400 mmHg for 2 min had the lowest mean GFP expression levels. This modified vacuum agroinfiltration approach utilized only distilled water and A. tumefaciens as components of the infiltration solution had substantially reduced the chemical cost. In conclusion, a simple and cost-effective laboratory-scale vacuum-assisted agroinfiltration system has been developed for recombinant protein production in planta. 2014/04/17 - 22:20

Earlier investigators have reported early Oral Contraceptives Pills (OCPs)
to be hepatotoxic. Despite the modifications on early OCPs in term of content
and dosage to lessen their side effects, paucity of literatures existed on the
effects of newer OCPs on liver function and integrity. This study is designed
to investigate the effect of OCP containing low doses of synthetic hormones
on liver function and integrity. The study involves 15 adult female rabbits,
divided into three groups (A, B and C). Group A serves as the control, while
B and C served as the test groups involving lower and higher weight rabbits
respectively. Throughout the period of the study water, rabbit feed and grasses
were given ad libitum to animals in each group. In addition, animals
in group B and C received for 7 and 14 days; 0.14 and 0.3 mL kg-1
b.wt. of OCP, respectively. At the end of the experiment, blood sample was collected
for the analysis of liver function parameters. The results showed progressive
increase in all the parameters studied in test group (B and C) compared to control
(group A). Specifically, there were significant increases in total bilirubin,
AST and ALT between the groups. While group B presented a non significant increase
in direct bilirubin, total protein and ALP compared with control values, group
C presented significant increase in these parameters compared to control and
test group B. The results of this study therefore suggest that OCP regardless
of their dosages, have potentials for impairing liver function and may induces
liver damage. 2014/04/17 - 22:20

Various plants are used in herbalism because of its phytochemical constituents
which have been proven to have medicinal effects by rigorous science and approved
by regulatory agencies. In the present investigation an attempt has been made
on qualitative and quantitative analysis of flavoniods from the leaf and flower
extract of Tagetes erecta. After the confirmation for the presence of
flavonoids in both the samples (Leaf and Flower) by qualitative analysis, quantitative
analysis (Ammonium chloride method) was carried out. In the present investigation
the recorded flavonoids ranged from 9.2 to 24.5 μg mg-1 in the
leaf sample and from 7.3 to 30.4 μg mg-1 in the flower sample,
respectively. 2014/04/17 - 22:20

Because of the diminishing fossil fuel reserves and increased CO2
accumulation in the atmosphere due to their burning, biofuels have been looked
as an alternative for sustainability and protecting the environment. Use of
plant byproducts such as lignocellulosic material as feedstock would be a viable
strategy avoiding the use of food (sugar) for bioethanol production. Lignocelluloses
are the most abundant resource in soil, its susceptibility to enzymatic digestion
favor the release of simple sugars which could be fermented to produce ethanol.
The cellulolytic enzymes that hydrolyze cellulosic feedstocks economically are
critical to develop the biofuel energy sector. Numerous highly effective pathways
for degrading biomass have evolved but enzymes for effective digestion of biomass
have been characterized only from few culturable organisms. These biomass-degrading
organisms are ill suited for genetic engineering or industrial applications
while conventional methods for identifying and cloning their individual enzymes
are inefficient. The discipline of metagenomics as the culture independent genomic
analysis of entire microorganisms in a particular environmental niche was evolved
as an effort to discover novel microbial enzymes to improve biomass utilization.
This review, focus on metagenomics as a promising approach for biofuel research
and its application in the NGS era. 2013/07/15 - 16:44

Palm Oil Mill Effluent (POME) cause serious pollutions of soil, water and environment
but could be exploited as substrate for microbial citric acid production because
of its rich mineral and high carbohydrate contents. Therefore in this study,
Aspergillus niger ATCC 9642 was grown in the different concentrations
(25, 50, 75 and 100%) of cellulase-digested POME for 7 days. Result showed that
citric acid was highest (0.78±0.02 g L-1) in the 75% POME
but lower (0.32±0.04 g L-1) in the undiluted POME. When supplemented
with methanol (3%), citric acid production increased (1.02±0.03 g L-1)
only in the 100% POME. Citric acid was not detected in the 25% and the non-digested
POME. Biomass measurement showed that the 100% POME gave the highest (10.2±0.6
g L-1) mycelial weight among the digested treatments. As the dilution
increased, the biomass concentration decreased proportionally. Biosolids from
the non-digested POME supplemented with glucose (1%) weighed 13.4±0.15
g L-1. The Chemical Oxygen Demand (COD) of the digested POME (100%)
dropped by 47.4% but the presence of methanol did not significantly affect the
COD kinetics. However, at higher POME dilutions, COD reduction rate was greater
than 63% but in the non-digested batches, COD reduction was negligible. The
sugar utilization patterns showed that glucose was completely used up in the
different POME dilutions except the undigested and glucose (1%) supplemented
batches which had 75 and 40% sugar uptake, respectively. The use of POME hydrolysates
as substrate could lower the present cost of citric acid as well as save treatment
costs and help solve environmental problems. 2013/07/15 - 16:44

In this review most of statistical tools currently applied in the bioprocess area were discussed and classified. The main three categories were: fair comparison of results, mathematical modeling for little studied systems and taking advantage of large volume of data for enhance robustness and efficiency. For each statistical technique, an example from literature was commented to demonstrate its utility in bioprocess problems. Besides, others statistical tools without a wide application, at the moment, in bioprocess were also discussed, taking into consideration the near future use of them. As a conclusion, a chart was constructed for guiding researchers to select the correct statistical technique according to the specific bioprocess problem. 2013/06/18 - 08:41

Marine sponge Aaptos-aaptos is known as antimicrobial compound aaptamine and its derivatives producer. To establish whether associated bacteria were the truly produced bioactive compounds, for preliminary, study of antimicrobial properties was done to know the potency of antimicrobial production in various carbon source. In this research, approximately 10 bacterial colonies were isolated from sponge Aaptos aaptos. Among isolated bacteria, colony Sp.2.11 was the most potential producing antibacterial compound. The partial gen 16S-rDNA sequencing of this bacteria showed 99% similarity with Rhodobacteracea bacterium species. Extra and intra cellular bacterial extract of this strain strongly inhibited the growth of pathogenic bacteria Staphylococcus aureus and Vibrio eltor while moderately against Bacillus subtilis. Optimization of activity was conducted by culturing Rhodobacteraceae bacterium in various carbon sources. Anti-gram positive bacterial production of extra cellular bacterial extract was highest when culturing this bacteria using minimum carbon source containing pepton, yeast extract in sea water medium (SYP), while anti gram negative bacterial was maximum in SYP medium with glucose as carbon source. Anti-gram positive bacterial production of intra cellular bacterial extract was highest in SYP with glycerol as carbon source, while anti-gram negative was highest in SYP with glucose as carbon source. In this research the highest biomass production didn’t increase the antibiotic production. 2013/06/18 - 08:41

Every year thousands of tones of Bolti fish (Tilapia nilotica) viscera are either discarded leading to environmental pollution and wastage of bioresources. This biomass has however a potential to generate considerable revenue and can be turned into a commercially viable business. The objective of this study was to evaluate the proximate protein characterization of the aqueous soluble phase of acidified and autolyzed Bolti fish viscera. Solubilization of fish viscera at natural pH (6.2); slight acidified pH (5.2) as well as acidified pH (4.2 and 3.2) was determined. Autohydrolysis reactions were conducted on freshly thawed viscera utilizing gradient temperatures and terminated at various time points (0, 8, 18, 32 and 47 h) by heat inactivation of the endogenous enzymes. Viscera were then characterized with respect to total nitrogen content, soluble protein, ammonia content, amino acid compositions and protein molecular weight distribution. Interestingly, endogenous enzyme activities were sufficient for obtaining a solubilization yield protein of up to 30 g L-1. SDS-PAGE profile showed that no characteristic band can be individualized for each hydrolysate and at the same time, greater variation was found only in the hydrolysate digested under initial pH 4.2 for 47 h. Amino acid analysis revealed that several amino acids were present in considerable amounts in the obtained hydrolysates, among them alanine, glutamic acid and proline. In addition, many hydrolysates contained useful quantities of the essential amino acids such as leucine, isoleucine, phenylalanine, histidine and valine and that may represent an interesting approach to upgrade these byproducts for food or feed purposes. In fact the results of our research would provide the incentive for commercial developments leading to large-scale and cost-effective production of fish peptone hydrolysates rich with essential amino acids from Bolti fish viscera. 2012/07/05 - 04:29

An efficient protocol was developed in vitro micropropagation of Gloriosa superba by using corm bud explant. MS basal medium supplemented with different concentration and combination of 2,4-D (2,4-dichlorophenoxy acetic acid), IAA (Indole-3-acetic acid), BAP (6-benzyl aminopurine), GA3 (Gibberellic acid), Zen (Zeatin), NAA (α-naphthaleneacetic acid), AC (Activated Charcoals) and CW (Coconut Water) were used. The 98.30±0.84% of yellowish Callus initiation was observed in MS media with (2, 4-D 1.0 mg L-1, IAA 0.5 mg L-1) after 4 week culture. This corm bud callus transferred to the shoots initiation medium. The maximum number (94.00±2.92) of multiple shoots was obtained on half strength of MS medium with (Kn (Kinetin) 1.0+BAP1.5+20% CW). The shoot lets transferred to the roots initiation medium. The 96.20±2.59% of multiple roots was obtained at the concentration of MS medium with BAP(8.0)+GA3(1.0)+Zen(0.5)+NAA(1.0)+2 g L-1 AC. In other case without addition of activated charcoal in the MS medium, only 92.20±1.92% of root initiation was occurred, 16% of root induction depends on the addition of activated charcoal present in the MS medium. The rooted plantlets were transferred into small plastic nursery tray which was containing vermi compost, sand and red soil in the ratio of 1:2:2 and kept in a mist house. After acclimatization in the mist house for 2-months, the regenerated plantlets were hardened in the greenhouse and successfully transferred into soil which shows 90% survival rate. This new protocol was standardized for easy mass propagation of such an endangered medicinal plant G. superba by using corm bud explants. 2012/07/05 - 04:29

Bacteria have potential ability to decolorize synthetic commercial dyes used for textile dyeing. Effluents from textile and dyeing industries cause serious pollution of soil, water and environment. Therefore, this study was aimed to isolate potential dye degrading bacteria from textile effluents and to evaluate their capability to decolorize commercially used textile dyes. Effluent samples were collected from different discharge locations of one selected textile industry. Six bacteria were isolated using a modified nutrient media. Results of biochemical test parameters of these bacteria matched with Bacillus subtilis (isolate B2, B7 and C1), Bacillus megaterium (isolate D3), Erysipelothrix (isolate C4) and Amphibacillus xylanus (isolate A1). These isolates were cultured with three different concentrations of seven different textile dyes viz. Cibacron red FN-R, novacron blue, terasil green, novacron navy, novacron orange and novacron yellow. Degradation ability of dye stuffs by the isolates (0.01, 0.05, 0.1 mg L-1) was observed by dye decolorization assay. Almost all dyes except novacron red were decolorized up to 99% by bacterial isolates after 3 days of incubation. 2012/07/05 - 04:29

Microbial Fuel Cells (MFCs) have attracted considerable attention over the last decade as a sustainable technology for electricity generation. Although, the concept of electricity production from bacteria was conceived nearly a century ago, only recently the technology has been sufficiently improved to make it useful as a method for energy generation. Performances of constructed MFCs with different enriched environmental samples were investigated under batch mode of operation after 24 h of incubation using different cathodic electrolytes and electrodes in mediator-less and with methylene blue as mediator. The maximum power density obtained was 48.85 mW m-2 with graphite rod and 42.59 mW m-2 with copper electrode and current density obtained was 108.57 mA m-2 with graphite rod and 88.01 mA m-2 with copper electrode all obtained with KMnO4 solution. 2012/07/05 - 04:29

Plantlets regeneration through callus has been achieved in medicinal plant Aerva lanata of Amaranthaceae. Callus obtained by culturing young leaf discs on Murashige and Skoog-1’s (MS) medium with 1.0 mg L-1 2, 4.D (dichlorophenoxyacetic acid) was subcultured on MS medium containing 2.0 mg L-1 6, Benzyl Amino Purine (BAP), produced 30-40 multiple shoots. Rooting was achieved on half or full strength MS medium with 0.5-0.1 mg L-1 Naphthalene Acetic Acid (NAA) in 14 days. Regenerated plantlets were acclimatized and transferred to soil were showed normal morphological characteristics. Survival of transplants was 90-95% under green house condition. 2012/07/05 - 04:29

Helicoverpa armigera (American bollworm) is a severe pest of many economically important crops such as cotton, pigeon pea, chickpea and tomato in the Indian subcontinent. Insecticidal cry genes from the bacterium Bacillus thuringiensis (Bt) have been used for developing transgenic crops. Transgenic cotton expressing cry1Ac gene has shown good level of protection from H. armigera. However, there is a threat of eventual development of resistance in insects upon large-scale cultivation of transgenic crops. Therefore, prospecting of Bt strains for new types of cry genes is desirable. This study was undertaken for screening of native Bt isolates for the presence of cry1Aa,b,c-type genes with the objective of cloning, sequence analysis, expression and evaluation of toxicity. Sixty three native Bt isolates recovered from different soil and grain samples from diverse agricultural and non-agricultural locations in India, along with 10 known Bt strains used as reference, were screened for the presence of full length cry1A-type genes by Polymerase Chain Recation (PCR) using a set of primer. The full length gene was obtained in the native Bt isolate SK-711 recovered from Red gram field, Lam, Guntur Andhra Pradesh and in 3 Bt strains. The gene from the native Bt isolate was cloned into an E. coli expression vector. The sequence of the cloned gene (GenBank accession No. GQ866913) was analyzed by comparison with previously known cry1A-type genes and was assigned the name cry1Ac33 by the Bacillus thuringiensis Nomenclature Committee. The gene was expressed in E. coli and evaluated for toxicity towards H. armigera. The cry1Ac33 gene, cloned from a native Bt isolate, has been found to be more toxic towards H. armigera than the holotype cry1Ac1 used as a control, based on LC50 toxicity analysis. 2012/03/12 - 21:01

The present study was aimed to purify and characterize extracellular lipase from Trichosporon asahii MSR54. An extracellular 27 kDa lipase from T. asahii was purified by anion exchange chromatography with 2.14 purification fold. It was magnesium stimulated. Reversible inhibition of EDTA was found to be by adding 15 mM magnesium chloride confirming its metallo nature. The temperature and pH optima for activity was 40°C and pH 9.0, respectively. It’s Km and Vmax was found to be 55.5 μM and 6.66 mM min-1, respectively using p-nitrophenyl palmitate as a substrate. The present enzyme hydrolysed a large array of oils and triacylglycerides with better specificity on corn oil and triolein, respectively. It was 1,3-regioselective during hydrolysis of triolein in aqueous as well as in micro-aqueous environment. It was stable towards most of the polar and non-polar solvents including methanol, DMSO, benzene. The present enzyme yielded 87.6% conversion of coconut oil to biodiesel after overnight incubation at 45°C. 2012/03/12 - 21:01

Comet assay is a sensitive, simple and well validated tool for measuring DNA damaged level in single cell. The assay is based on embedding cells in a thin layer of low melting agarose, which is located on the microscopic slide. Comet assay is a method used as an assessment tool for early biological effect biomarker to the occupational exposure. This investigation evaluated the occupational exposure effects in the workers acting as an enhancement factor of ageing. In order to do this assessment 120 workers as exposed and 120 controls as non-exposed were considered. DNA damage was evaluated by measuring the extent of DNA migration in buccal cells. It was found that the mechanical workshop workers who were occupationally exposed to the genotoxic agents for different periods of time recorded significant increased in the levels of DNA damage as compared to the controls. The data were considered for assessment of their correlation with early ageing. The investigation indicated that the responses of DNA damage among the exposed people in the work places could be related to confounding factors such as life styles as well. Comet assay in the evaluation of DNA damage reflected the sensitivity of this method at moment of collecting samples. 2012/03/12 - 21:01

Sago palm (Metroxylon sagu) is an emerging important food plant for marginal lands. The palm reproduces in the wild slowly by means of suckers. Thus, in vitro culture methods must be employed to aid its multiplication. In this study, varying concentrations of auxin and cytokinin were evaluated to establish appropriate media for shoot regeneration of sago palm (Metroxylon sagu Rottb.) leaf explants. Shoots that regenerated from modified Murashige and Skoog (MS) medium containing 10 mg L-1 2,4 D and 0.2 mg L-1 kinetin with 0.3% activated charcoal, 3% sucrose and 2 g L-1 gelrite at pH 5.7 showed the greatest weight increases. However, shoot explants inoculated in media adjusted to pH 6.8 showed faster morphological changes. 2012/03/12 - 21:01

Extraction of good quality DNA from Nothapodytes nimmoniana, an important anti cancerous medicinal plant containing many secondary metabolites, was a challenging task as many of these compounds co-precipitate with DNA and render the DNA unsuitable for downstream applications. Previously reported protocols yielded low quality brownish DNA, that was not amenable to further analysis. In this study, protocol was optimized with cetyltrimethylammonium bromide (CTAB) for isolation of genomic DNA. Important modifications in the optimized protocol included composition of homogenization buffer, volume of tissue: buffer ratio, modification of both extraction and precipitation temperature and duration. The efficiency and reliability of the method was tested by assessing quantity and quality of the extracted DNA by restriction analysis and Polymerase Chain Reaction (PCR) using Inter-Simple Sequence Repeat (ISSR). The yield of DNA ranged from 10-250 μg g-1 of leaf tissue. The PCR protocol comprising 50 ng template DNA, 0.5 mM dNTPs, 0.5 U Taq DNA polymerase and 0.5 μM primer concentrations found optimum for carrying out ISSR analysis in N. nimmoniana. The present optimized protocol for DNA isolation and ISSR technique serve as an efficient tool for carrying out further molecular studies in this species. 2012/03/12 - 21:01

Recent breakthrough in industrial biotechnology offer important economic opportunities for the utilization of agro-industrial residues such as Banana Peels (BP). BP as a complex biological material is an important by-product of several cottage and major hospitality industries. Chemically, it contains cellulose, hemicelluloses, lignin and simple sugars. Due to its availability and value, it is capable as an ideal substrate for microbial process for production of valued-added products. Several efforts have been made to produce protein enriched animal ration, industrial enzymes, citric acid and other industrially viable products. Pre-treatment operations have resulted in improved substrate utilization by microorganisms. Solid State Fermentation (SSF) and submerged state fermentation (SmF) are two promising techniques majorly employed for bioconversion of BP for its value addition. The article reviews recent developments in process and products developed for valorizing BP through biotechnological approach. Information on most recent applications and developments are carefully documented. 2012/02/23 - 06:22

Relationship between fungal growth morphology and the ability to produce lipase in solid state fermentation was studied employing four fungal strains (A. niger DSMZ 2466, A. niger SDTC SRW-4, T. viride SDTC EDF 002 and A. terreus DSMZ 5770). The fermentations were performed in 500 mL flask using palm kernel cake as substrate. Scanning Electron Microscope (SEM) was used to examine growth morphology of the fungi. Pearson correlation and regression analyses performed on quantified morphological dimensions and the fungal growth, enzyme productivity and total protein were able to generate predicted values of equation models. All four strains showed good growth on palm kernel cake at day 6 of fermentation ranging from 0.49 to 0.71 mg g-1 dry substrate. Among these strains, substantial lipase activity was only obtained by A. niger DSMZ 2466 and A. niger SDTC SRW-4 which also showed positive relationship between spore diameter (quadratic, R2; 0.999) and branch diameter (linear, R2; 0.971) with lipase activity. T. viride SDTC EDF 002 and A. terreus DSMZ 5770 showed positive correlations for tip length and tip diameter and size of hyphae diameter and the total protein content in the fermentation substrate. Results from this study showed that the fungi with larger spore diameter and branch diameter contributed to higher ability in producing lipase. 2012/02/23 - 06:22

Genetic relationship among 15 citrus fruits (one from the genus Poncirus and the rest from the genus Citrus) was assessed using Randomly Amplified Polymorphic DNA (RAPD) markers. Seven primers were selected after screening of 39 decamer primers which generated 91 clear and bright polymorphic bands. The Unweighted Pair-group Method using Arithmetic averages (UPGMA) dendrogram based on Nei’s genetic distance segregated 15 citrus fruits into 2 main clusters where Poncirus trifoliata alone formed one cluster and the rest 14 species grouped together into another cluster. Inter-species similarity index and the UPGMA dendrogram based on Nei’s genetic distance revealed that Citrus megaloxycarpa and C. limonia as the most closely related species among the 15 citrus fruits under study. 2012/02/23 - 06:22

Microalgae are being studied aggressively lately as a potential feedstock for biofuel and other valuable compound production. In this study, marine microalgae were collected at Fisheries Department of Terengganu in Malaysia which were then being isolated, purified and identified using rDNA approach. For the molecular identification analysis, several DNA extraction methods were investigated in order to obtain high quality algal DNA. Chloroplast genomes were amplified successfully using eight sets of universal chlorophyte primers from the genomic DNA extracted. These amplified fragments were then purified and ligated into pGEM-T vector which was subsequently cloned into E. coli competent cell. Plasmids DNA of the selected transformed cells were extracted and prepared for sequencing. The sequences were analyzed and compared with other strains in NCBI database. Based on the chloroplast sequences, the collected strain is identified as Parachlorella kesslari. As for the analysis of biochemical composition of this strain, the result revealed that it composed of 30% lipid, followed by 25% carbohydrate, 15% protein and 8% starch, respectively. Forty percent of the extracted lipid was composed of PUFAs through GC-MS analysis. 2012/02/23 - 06:22

Pogostemon cablin also known as Patchouli belongs to the family Lamiaceae
is an important aromatic plant producing patchouli oil from its leaves extraction.
The main objective of this study was to develop an effective method for in
vitro propagation of Malaysia cultivated P. cablin. The effect of
varying the strength of solid MS medium was investigated using nodes explants.
Different concentrations of 6-Benzlaminopurine, BAP (0, 0.25, 0.5, 1.0, 2.0
mg L-1) and alpha naphthalene acetic acid, NAA (0, 0.25, 0.5, 1.0,
2.0 mg L-1) were added to both full and half strength MS (Murashige
and Skoog) medium for regeneration of this plant. After 4 weeks, the shoots
regeneration from the nodal segments was obtained in all medium. Then, clumps
of shoots (5 shoots) were separated and transferred to five MS medium concentrations
(0.25 mg L-1 BAP, 0.5 mg L-1 BAP, 0.25 BAP/0.25 NAA and
1.0 BAP/0.25 NAA). MS supplemented with 0.25 mg L-1 BAP alone gave
the highest number of shoots (32.93±3.93) and length (3.80±0.27
cm) followed by MS supplemented with combination of 1.0 mg L-1 BAP
and 0.25 mg L-1 NAA hormones. There was also a significant effect
of hormones and media strength on the number of shoots obtained. The statistical
analysis done showed that there was very high interaction between both factors
of hormones concentration as well as the medium strength. In the other words,
the number of shoots obtained to hormones concentration depends on medium strength
and vice-versa. In conclusion, in vitro regeneration of P. cablin
from node explants in MS medium supplemented with BAP or NAA was successfully
obtained. The healthy propagules that were obtained in this research can be
used for further procedure of acclimatization. 2012/02/23 - 06:22

The present study deals with the standardization of different cultural conditions for extracellular α-amylase production by thermophilic Streptomyces sp. MSC702 in submerged fermentation (SmF). The appropriate incubation period (48 h), temperature (50°C) and pH (7.0) were determined. The effects of derived and natural carbon sources, inorganic and organic nitrogen sources were also examined. Maximum α-amylase production i.e., 435.71 and 373.89 U mL-1 were achieved by employing derived (D-inositol) and natural (rice bran) carbon sources, respectively. Among the tested nitrogen sources, ammonium sulphate and peptone were found the best inorganic and organic sources, respectively. The C:N ratio found to be the optimum was 1:1. The highest α-amylase activity (807.64 U mL-1) was obtained by utilizing rice bran and wheat bran in 1:2 ratio as the substrate with supplements of D-inositol (1% w/v), ammonium sulphate (0.5% w/v) and peptone (1% w/v). By using the optimized cultural conditions with further characterization, this α-amylase may be utilized in wide spread applications like detergent, saccharification and pharmaceutical industry. 2011/12/03 - 13:05

The yield of banana and plantain which is an important food is affected especially by black leaf streak disease. Pollen from resistant wild types is used by conventional breeding to create new hybrids resistant to black leaf streak disease. A strategy of pollen genetic resource preservation and storage is necessary for this breeding. The main objective of this study was to optimize the conditions for in vitro germination, conservation and storage of pollen from three banana diploids (Musa acuminata). Pollen grains used in this study were collected from Njombe and Ekona banana collections. The basal media used for in vitro culture were those of Heslop-Harrison and Brewbaker and Kwack. Different temperature and pH values tested allowed for the evaluation of germination rates. Pollen were desiccated and stored in a freezer (-20°C) and in a refrigerator (+10°C). The results obtained showed that the basal medium of Heslop-Harrison supplemented with 5% sucrose allowed to obtain optimum pollen germination rate. The optimum temperature was 30°C and the optimum pH values were 6.5 for Calcutta and 6.8 for M53 and Zebrina. The pollen grains of the three varieties can be conserved freshly in the refrigerator at +10°C for at least two months. These results are very important for banana and plantain breeding programs because the conditions of pollen storage for the three varieties are well established and permit genetic material sharing and diffusion between different banana research centers. 2011/12/03 - 13:05

An efficient protocol for in vitro propagation of Terminalia arjuna is described by callus regeneration. Four different explants were used to establish callus to develop shoot and root regeneration method. MS (Murashiege and Skoog), LS (Linsmaier and Skoog) and B5 (Gamborg) basal medium supplemented with 2,4-Dichlorophenoxyacetic acid (2,4-D, 0.1-20.0 mg L-1), with combination of Naphthalene acetic acid (NAA 0.1-20 mg L-1), Indole-3-acetic acid (IAA 0.1-20 mg L-1) and Benzylaminopurin (0.1-20 mg L-1). Ms medium was found to be the most favorable for callus induction compare with LS and B5 media. Maximum number of callus regeneration was obtained on MS medium containing 2,4-D 3.0 mg L-1. The callus culture to develop the shoot and root initiation in MS basal medium substituted with 5 mg L-1 2, 4-D+0.01 mg L-1 Kinetin and 1.0 mg L-1 Gibberellic acid (GA3). The rooted shoot plantlet was transferred in to small plastic cups containing sterile vermiculate, sand and red soil in the ratio of 1:2:2 and were kept in a mist house. The regenerated plantlets were hardened in the greenhouse and successfully transferred in soil with 87% survival rate. This in vitro micro-propagation method with possibility of developing a new protocol was standardized the T. arjuna plant. 2011/12/03 - 13:05

Genotoxic effects of hazards at the environmental and occupational exposure contribute to ageing acceleration. These compounds induce genetic damages in the exposed cells such as buccal cells. The study was carried out on the mechanical workshop workers to investigate micronuclei (MN) formation in the exposed subjects. The formation in the cells was evaluated by using MN test. The applied buccal cells as sensitive sources showed the genotoxic and cytotoxic damages which could accompany ageing. Difference in MN frequency between the workers and controls was statistically significant (p = 0.001). It showed statistically significant difference between the older and younger groups in the workers and controls (p = 0.001) as well. The results showed that the occupational exposure increased MN frequency in the workers. It was concluded that the induced genomic damages could act as an accelerating factor to enhance the speed of ageing in the workers. 2011/12/03 - 13:05

This study was carried out in order to determine the effect of three test rations comprising Brachiaria ruziziensis grass silage and sawdust/poultry litter silage which replaced some conventional feed ingredients (wheat offal, soya bean cake and maize offal) at varying levels in the diets of goats. Twelve young West African dwarf does with mean ages and weights of 6±0.17 months and 4.2±0.44 kg, respectively, randomly divided into three groups of four replicates were used for the study which lasted for forty-five days. The animals were fed three test rations with sawdust/poultry litter silage replacing 50% (Diet 1), 25% (Diet 2) and 0% (Diet 3 or Control. In addition, all groups received Brachiaria ruziziensis grass silage at 5% inclusion level. Parameters measured include Production attributes, Haematology and serum biochemistry. Results showed that the haematological and serum biochemical parameters showed no significant differences (p>0.05) among groups. However, significant differences (p>0.05) existed in the production attributes with respect to mean final gain, mean daily gain, mean daily intake, mean total feed intake and apparent protein digestibility, feed conversion efficiency, but not in their rumen motility. Cost of production was significantly (p<0.05) lowest in diet 1 compared to diets 2 and 3, respectively. It was observed that sawdust/poultry litter silage was most beneficial at 25% inclusion level as animals fed this diet had significantly higher weight gain and better feed conversion efficiency compared to those of 50% and Control groups. 2011/12/03 - 13:05

The aim of this study was to clone and optimize production of a recombinant antibody against human leptin receptor which showed inhibitory effects on leptin signalling. DNA sequence encoding Fab fragment of a mouse monoclonal antibody was cloned into pComb3 vector. E. coli was transformed with the plasmid and recombinant antibody was produced. In order to achieve high level of protein expression, recombinant antibody production was performed using different E. coli strains and culture conditions. Antibody production was detected using ELISA (Enzyme-linked immunosorbent assay), Western blotting and Dot blotting techniques. Recombinant Fab fragment of antibody was produced in E. coli successfully at reasonable quantities with comparable affinity to the antigen compared to antibody produced by original hybridoma cells. The results showed that among different culture media, SB medium was the best medium and among different temperatures 22°C was the best temperature for recombinant antibody production. From three different E. coli strains (JM109. XL1-Blue and BL21) tested, JM109 produced the highest levels and among different IPTG concentrations 1 mM induced the highest level of antibody production. 2011/07/27 - 01:06

This study was conducted to assess microbiological quality of a local fresh water fish of Bangladesh named mola, Amblypharyngodon mola. The effects of freezing temperature, irradiation and their combination treatments on the associated microorganisms during storage of the fish were also evaluated. For this purpose, fish samples were collected from a local market and irradiated with different doses (2.5, 5.0, 7.5 and 10.0 kGy) of gamma radiation and kept at -20°C for 6 months followed by microbiological assay after each month interval. The average Total Viable Bacterial Count (TVBC), Total Staphylococcal Count (TSC), Total Coliform Count (TCC), Total Faecal Coliform Count (TFCC), Total Aeromonas Count (TAC) and Total Fungi Count (TFC) of fresh mola fish samples were 9.74x1007, 3.60x1006, 9.70x1004, 6.40x1005, 4.00x1005 and 2.30x1003 cfu g-1, respectively. Seventy six bacterial strains were isolated and identified from these samples which include Staphylococcus aureus, Micrococcus varians, Aeromonas hydrophila, Pseudomonas aeruginosa, Escherichia coli, Klebsiella ozaenae, Bacillus subtilis, Bacillus megaterium, Klebsiella edwardsii and Micrococcus radiodurans. When treated with gamma radiation dose, the total coliform, faecal coliform, Aeromonas sp. and fungi of the samples were eliminated at 2.5 kGy and total viable bacteria and staphylococci were eliminated at 7.5 kGy. During frozen storage, microbial counts were gradually decreased with time. However, the combination of these two treatments was found more effective than any of the single treatment for elimination of the associated microorganisms. The moisture contents of mola fish were not changed significantly at different stage of storage periods and also at different radiation doses. On the other hand, losses of total protein contents were also very low with increasing storage time and radiation dose. The findings indicated that, the combination treatment can be applied for long time preservation of mola fish in Bangladesh without any significant loss of moisture and protein values. 2011/07/27 - 01:06

Biochemical Oxygen Demand (BOD) is an important parameter for water pollution monitoring. Since the conventional BOD assay by American Public Health Association (APHA) takes 5 days to complete (termed BOD5), development of an alternative method that can rapidly measure the amount of biodegradable organics in wastewater is highly desirable. In the present study, single and mixed microbial cultures isolated from different environments in Malaysia were tested as potential biocatalyst in an assay called Ferricyanide Mediated-Biochemical Oxygen Demand (FM-BOD) used for rapid BOD detection. Unlike the BOD5 assay, the FM-BOD is based on biochemical degradation of organic compounds via reduction of ferricyanide as an alternative terminal electron acceptor to dissolved oxygen. The increase in concentration of microbially produced ferrocyanide during incubation of the microorganisms in the presence of ferricyanide and organic substrates was measured voltammetrically using a 10 μm platinum working electrode. Measurements were conducted using linear sweep voltammetry at potential 0-600 mV versus Ag/AgCl reference electrode. Limiting currents were determined by voltammetry at Eapp = +450 mV (vs Ag/AgCl). The amount of ferrocyanide produced was found to be proportional to the amount of biodegradable organics. From the study, it is shown that mixed microbial consortium acted as a better biocatalyst for rapid BOD detection compared to both single cultures and a commercial consortium. The microbial consortium consists of four species of microorganisms. FM-BOD assays were conducted in optimized conditions with microbial consortium at final absorbance of OD600 2.5 and ferricyanide at concentration 60 mM. Results showed that the FM-BOD assay was able to detect biodegradable organics in 1 h compared to the current BOD5 assay of 5 days. This FM-BOD assay with optimized conditions is applicable to food wastewater consisting of easily degradable carbon source. 2011/07/27 - 01:06

Vibrios are halophilic bacteria and are ubiquitous in marine environments. The diversity of these bacteria in tropical areas is not clearly understood. In this study, Vibrio spp. including Vibrio parahaemolyticus were investigated at the Andaman Tarutao Island, Thailand. Water samples were collected at Son Bay and Ruesri Bay located on opposite sides of the island. The total numbers and the predominance of Vibrio spp. were determined by cultivation on thiosulfate citrate bile- salts sucrose agar. Enumeration of V. parahaemolyticus was performed on CHROMagar Vibrio. There was no correlation between the total numbers of vibrios, V. parahaemolyticus, temperature and salinity detected in both areas. By cultivation, V. parahaemolyticus was regularly observed at Son Bay except in December whereas it was detected only in April and November at Ruesri Bay. The diversity of Vibrio spp. was investigated using the Denaturant Gradient Gel Electrophoresis (DGGE) technique. The diversity of Vibrio spp. detected at Son Bay was homogeneous whereas it was more heterogeneous at Ruesri Bay. Interstingly, in December although V. parahaemolyticus was not detected by cultivation; it was present in this month according to the DGGE technique. Thus, the number of this bacterium was probably low or it was in a non-culturable phase. In this study, V. harveyi was detected in both areas every month by DGGE. This indicates that this bacterium is part of the normal microbiota and is present in the Andaman Sea throughout the year. Correlation between copepods and the numbers of V. parahaemolyticus was not observed in this study. 2011/07/27 - 01:06

SE clan of serine proteases have Lysine, Serine and Tyrosine residues as catalytic triad in their active site which is an exception to all other serine proteases having conserved residues Serine, Aspartic acid and Histidine in their active sites. The aim of this study was to explore the structural distinctiveness of different S12 family (D-Ala-D-Ala carboxypeptidase B family) serine peptidases from diverse groups of species belonging to SE clan, using molecular modeling techniques. Amino acid sequences of those proteases from each group of organisms, i.e., archean, protozoan, fungal, plant and human were taken from the MEROPS database in FASTA format. Homology and threading modeling approach were used to construct all the structures by SWISS MODEL and LOOPP server respectively. Structural quality assessing validation programs (PROCHECK, MODELYN, MOLPROBITY and PROSA) of the final model have demonstrated its reliability for further studies. A full structural database was constructed starting from bacteria up to human, focusing on the catalytic triad of the proteases. MODELYN study showed statistically that the region of the catalytic triad is highly conserved from bacteria to human. Phylogenetic analysis also supports their evolutionary connection. MOLMOL study showed that the catalytic sites of all the SE Clan proteases exhibit acidic regions in the surface electrostatic potential maps but a few of them contain both large patches of positive and neutral potentials. Hence some of the proteases with predominant positive and neutral regions around the catalytic site can be used as potential drug target against bacterial and protozoan pathogens. 2011/07/09 - 12:22

Cold-active α-amylases provided a large biotechnological potential and offers numerous economical and ecological benefits through energy saving process. It also minimizes undesirable chemical reactions that could occur at high temperatures. The objective of present study was to isolate novel bacterial strain for production of cold-active α-amylase. On the basis of primary screening thirty cold-active α-amylase producing bacteria were isolated from soil of Gangotri glacier, Western Himalaya, India. The isolated bacteria were subjected to enzyme production and one potential isolate (GA2) was selected for production optimization. The α-amylase production was found maximum (5870 units) at 20°C and pH 9 after 120 h incubation. Among the carbon and nitrogen sources, lactose (1%) and yeast extract (1%) was found best source, respectively. The isolate GA2 (Gangotri amylolytic) was resistant to penicillin (10 μg) among tested antibiotics and as per plasmid curing results, amylase production was a plasmid mediated characteristic. The phylogenetic analysis revealed that GA2 have highest homology with Microbacterium foliorum (99%). This was the first report on cold-active α-amylase production by M. foliorum GA2 and on the basis of 16S rRNA sequences also got an accession number HQ832574 from NCBI. 2011/07/09 - 12:22

Evaluation of genetic diversity was made using RAPD as a genetic marker system in allohexaploid wheat (T. aestivum). A set of 30 RAPD markers were used for 43 wheat cultivars to examine their utility for detecting DNA polymorphism in genotypes and estimating genetic diversity among them. A total of 228 different RAPD bands were scored for the presence of bands among all the varieties. Out of 30 oligonucleotide primers used, 5 primers gave reproducible DNA product (bands) suitable for the establishment of genetic diversity among the varieties. The Genetic Similarity (GS) coefficient for all pairs of genotypes was ranged from 0.66 to 1.0. V 35 (HD 2747) and V 39 (HI 8498) had high coefficient of similarity (0.982) while, V 4 (VL 804) and V 6 (VL 809) showed less coefficient of similarity (0.646). It indicated that V 35 (HD 2747) and V 39 (HI 8498) carry closely related ancestry while the cultivars V 4 (VL 804) and V 6 (VL 809) are not close to each other. Present results revealed the potential of RAPD technique in determining genetic diversity and make suitable fingerprints for wheat cultivars. 2011/07/09 - 12:22

The objective of the present study was to determine the effect of genotype and culture media in anther culture of boro rice hybrids. Anthers from three f1 hybrids of boroxhigh yielding indica rice varieties GautamxBPT 5204, Krishna HansaxNDR 359 and IR64xNDR 359 were cultured on modified SK media and modified N6 media. Modified SK media was found to be more suitable for callus formation in all the f1 hybrids. The cross GautamxBPT 5204 showed maximum callus induction frequency followed by Krishna HansaxNDR 359. Only the cross of Krishna HansaxNDR 359 was responsive to green plant regeneration while IR64xNDR359 and GautamxBPT 5204 resulted in only albino plants. Anther culture response is influenced by the genotype and media composition in indica rice hybrids. 2011/07/09 - 12:22

The aim of this study was to purify and characterize citric acid cycle enzyme malate dehydrogenase (MDH; EC 1. 1. 1. 37) from Pseudomonas aeruginosa. The purification steps consisted of ammonium sulphate precipitation, ion-exchange chromatography and gel filtration. A typical procedure provided 638-fold purification with 23.0% yield. Single band was observed in both native gradient-and SDS-PAGE. The molecular weight estimated for the native enzyme was 70.5 kDa whereas subunit values of 36 kDa were determined. Hence, MDH is a dimer of identical subunits. The enzyme was highly active at pH 8.0 when NADH was used as the cofactor and was highly stable at pH 7.0. The optimum temperature for the enzyme activity was recorded to be 40°C. Oxaloacetate was determined as the specific substrate with an apparent km of 10 μM. The characteristics of thermo-stability and its high activity at alkaline pH suggest its potential diagnostic, therapeutic and beverage related applications. This MDH may be of value in developing a serological test for pneumonia which is caused by P. aeruginosa. 2011/07/09 - 12:22

The present study has evaluated the genetic variability among three salt marsh plant belongs to the family of Chenopodiaceae using random amplified polymorphic DNA markers. The method involves a modified Cetyl trimethylammonium bromide extraction, using polyvinyl pyrrolidone while grinding, successive long-term Chloroform: lsoamyalcohol extractions, an overnight RNase treatment with all steps carried out at room temperature. The yield of DNA ranged from 1.5-2.5 μg μL-1 per gram of the leaf tissue. The technique is ideal for isolation of DNA from different salt marsh plant species and used for Randomly Amplified Polymorphic DNA (RAPD) analysis. RAPD protocol was optimized based on the use of lower concentrations of primer (2 μM) and Taq polymerase (2 units), 50 ng of template DNA, higher concentration of MgCl2 (2 mM) and an annealing temperature of 37°C, resulted optimal amplification. The results suggest that the optimized protocol for DNA isolation and PCR was appropriate to diversity analysis of salt marsh plants. 2011/07/09 - 12:22

From time immemorial, man has remained dependent on plants for medicine. From a historical perspective, it is evident that the fascination for plants is as old as mankind itself. The plant kingdom represents a rich storehouse of traditional medicines and organic compounds that may lead to development of novel agents for various disorders. Moringa oleifera Lam (Syn Moringa pterygosperma Gaertn) commonly known by regional names such as drumstick tree, sajiwan, kelor, murungai kaai, saijhan and sajna, is a natural as well as cultivated variety of the genus Moringa belonging to the family Moringaceae. It is a small or medium sized tree, about 10 m high, cultivated throughout India. It is a multipurpose tree known as nature’s medicine cabinet. It is best known as excellent source of nutrition and natural energy booster. Different parts of this plant are being employed for the treatment of various ailments in the indigenous system of medicine. It possesses antitumor, antipyretic, antiepileptic, anti-inflammatory, antiulcer, antispasmodic, diuretic, antihypertensive, cholesterol lowering, antioxidant, antidiabetic and hepatoprotective activities. This plant have broad spectrum activities so, this review focuses on numerous economic application, distribution and commercial importance of Moringa oleifera along with its traditional medicine and culinary uses, so that it can be grown enormously and can be used for various indigenous purposes. Considering its relevance, further research is required to explore the potential from this medicinal tree. 2011/06/10 - 10:02

Small interfering RNA or silencing RNA plays a variety of roles in cellular process of organisms. Studies in bacteria and archaea had gene silencing pathways that are triggered by transgene expression or viral replication. RNA silencing mechanism is first recognized as antiviral mechanisms that protect organisms from RNA viruses, or which prevent the random integration of transposable elements. Clustered Regularly Interspaced Short Palindromic Repeats (CRISPRs) are arrays of prokaryotic DNA sequences that mediate a form of acquired immunity to specific viral pathogens. CRISPR-associated (CAS) proteins play an important role in the initial recognition of phage genetic material and incorporating these proto-spacers in the CRISPR array. Phage-derived spacers are incorporated at the CRISPR 5' leader sequence so that the ordered sequence of spacers essentially gives a temporal record of the infection history in that bacterial population. Hence, CRISPRs offer a high-resolution method for molecular typing of bacterial hosts and their pathogens based on the unique CRISPR signature. 2011/06/10 - 10:02