Vol. 6(1) February 2011
Random Mutagenesis on Wautersia Eutropha for Improving
Poly-β-Hydroxybutyrate Production from Inexpensive Carbon Sources
ADas Santanu, Sikdar Munmun and Veerasamy Muralikrishnan
Wautersia eutropha, a source of homopolymer and heteropolymers
of biodegradable poly-β-hydroxybutyrate [P(3HB)], a potential alternative to recalcitrant
plastics, was subjected to random mutagenesis using ultraviolet irradiation. Five
mutant isolates, namely Wautersia eutropha M1, Wautersia eutropha M2, Wautersia
eutropha M4, Wautersia eutropha M5, Wautersia eutropha M6 were obtained on screening
for poly-β-hydroxubutyrate yield. All five mutant strains and wild-type Wautersia
eutropha rendered better P(3HB) yield in minimal salt broth supplemented with surplus
fructose than nutrient broth. Wautersia eutropha M2 and Wautersia eutropha M5 were
screened for improved P (3HB) yield. Wautersia eutropha M5 efficiently utilized
cane molasses rendering a highest P(3HB) content of 57.42% at 20g.L-1 concentration
whereas the other two strains showed insignificant P(3HB) yield from molasses. P(3HB)
content increased with increasing concentration of sago industry liquid waste by
the screened strains with a highest P(3HB) yield of 56.67% at 30g.L-1 concentration
by Wautersia eutropha M5. Arrack gave least growth and poor P(3HB) yield by the
screened strains.
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Valuable Antioxidant and Antimicrobial Extracts from
Rhizophora Mucronata of Asiatic Mangrove Forests
Haq Imdadul, Sani Wirakarnain, Philip Koshy , Rafat Arash, Hossain A.B.M. Shariff
and Taha Rosna Mat
Abstract: The antioxidant
and antimicrobial properties of ethanol, methanol and chloroform extracts of Rhizophora
mucronata leaves were examined in this study. The antioxidant activities of the
samples were evaluated using a combination of enzymatic and non-enzymatic methods
namely superoxide dismutase determination, erythrocyte haemolysis protection
and 2,2-diphenyl-1-picrylhydrazil free radical scavenging assays. Folin-Ciocalteu
reagent method was used to estimate the amount of total phenolic compounds of the
extracts. Ethanol, chloroform and methanol extracts of the leaves showed the highest
antioxidant potential in superoxide dismutase, erythrocyte haemolysis and free
radical scavenging assays respectively. The highest total phenolic content was found
in ethanol extract followed by methanol extract. Paper disk diffusion method was
applied to determine the antimicrobial activities of ethanol and methanol extracts
of the leaves. Both ethanol and methanol extracts could inhibit the growth of Escherichia
coli, Staphylococcus aureus and Bacillus cereus while no inhibition was detected
against Pseudomonas aeruginosa.
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Effect of Corn Oil as a Vehicle on Kidney of Female
Swiss Albino Mice Mus Musculus by Gavages Method
Anjali S.,Tanuja, Nath A., Singh J. K. and Ravish K.
Abstract: The purpose
of this study was to investigate the effects of corn oil on histological and biochemical
parameter of kidney of Female Swiss albino mice. Mice were administered corn oil
at the dose of 10 ml /kg body weight for the period of six weeks. No morphological
alterations were observed. But fur soiling was observed among few mice of treated
groups during the experimental period. Histological examination of kidney of female
mice revealed mild alterations like tubular disarrangement. Few foci of hemorrhage
and fatty degeneration were observed only in four weeks and six weeks corn oil treated
groups. Mice of one week and two weeks group showed neither abnormal condition
nor any severe lesions in the kidney. Comparative study of treated groups of mice
with control group of mice was done. The serum biochemical test or kidney function
test (Urea and creatinine) were estimated by Fully Automated Biochemistry Analyzer
(Model No.-Selectra- E” (Vita lab) by Merck. These data showed that corn oil at
the rate of 10 ml/kg body weight can be used as a vehicle for toxicological studies
but may cause adverse effects on the renal tubules if used for prolonged periods.
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The Extraction Efficiency of Apigenin from Scutellaria
barbata comparing pressurized Liquid extraction and Soxhlet extraction
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Lee Hanna and Shim Sang Hee
Abstract: Pressurized
liquid extraction of apigenin from Scutellaria barbata was compared to the traditional
Soxhlet extraction using methanol. The conditions for pressurized liquid extraction
were as follows: the extraction solvent was 100% ethanol, 50% aq ethanol or water;
the extraction temperature was 120˚C; static extraction time was 5 min. For the
traditional Soxhlet extraction, the sample was boiled at 50˚C for three hours. According
to calculations by regression parameters using HPLC analysis, the extraction yields
of apigenin were 1.25 mg/g by pressurized liquid extraction (extraction solvent,
100% ethanol; temperature, 120˚C; static extraction time, 5 min), 0.32 mg/g by pressurized
liquid extraction (extraction solvent, 50% aq ethanol; temperature, 120˚C; static
extraction time, 20 min) and 0.88 mg/g by Soxhlet extraction using methanol. Pressurized
liquid extraction using 100% ethanol yielded 74.4 % higher apigenin than using 50%
aq ethanol and using 100% ethanol yielded 29.6% higher apigenin than using the Soxhlet
extraction technique.
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Antibacterial and Antifungal Activity of different
Lichens Extracts and Lichen Acid
Kosanić Marijana and Ranković Branislav
Antibacterial and antifungal activity of the acetone,
methanol and aqueous extracts of the lichens Lecanora atra, Parmelia reticulata,
Parmelia oomphalodes and salazinic acid from Parmelia reticulata lichen were explored
in vitro against six species of bacteria and eight species of fungi. The antimicrobal
activity was estimated by the disc-difusion method and determination of the minimal
inhibitory concentration (MIC) by the Broth tube dilution method. The aqueous extracts
of all of the tested lichens were inactive, whereas the acetone and methanol extracts
showed antimicrobial activity against the majority of the tested organisms. Among
the extracts, the strongest activity was found in the acetone extract of the lichen
Parmelia omphalodes where the least measured MIC value was 0.78 mg/ml. Salazinic
acid also showed strong activity against bacteria and fungi. The lowest measured
MIC value was 0.39 mg/mL related to the Klebsiella pneumonie species. The bacteria
were more sensitive related to the tested fungi. Generally, Bacillus subtilis and
Bacillus mycoides were the most sensitive of the tested bacterial species while
Botrytis cinerea and Candida albicans were the most sensitive fungal species. There
was no antimicrobial activity against Escerichia coli species. The results obtained
show that the tested lichen and their compound demostrated a strong antimicrobal
effect against the tested microorganisms.
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In Vivo Immunomodulating Activities of Ethanolic Leaf
Extract of Stevia rebaudiana in Albino Rats
Shukla Shruti , Mehta Archana, Kim Myunghee and Ayyadurai N.
Abstract The present
study was undertaken to explore the in vivo immunomodulating activities of ethanolic
leaf extract of Stevia rebaudiana (Bertoni) in albino rats. The immunomodulatory
activities were determined by neutrophil adhesion test, haemagglutinating antibody
(HA) titre, delayed type hypersensitivity (DTH) response and phagocytic activity.
The response produced by oral administration of ethanolic leaf extract showed
a significant increase in neutrophil adhesion and delayed type hypersensitivity
(DTH) response. With 300, 400 and 500 mg/kg/day doses of the extract, the DTH responses
of 24 h (mean ± SD % increase in paw volume) were found to be 0.41 ± 0.01, 0.50
± 0.01 and 0.58 ± 0.01 respectively as compared to the corresponding value of 0.18
± 0.01 for the control group. These differences in DTH response were statistically
significant (P<0.05). The augmentation of humoral immune response to sheep red blood
cells by ethanolic extract (200 - 500 mg/kg) was evident by increase in antibody
titres in rats. A dose-related increase in antibody titres was observed. The ethanolic
leaf extract also evoked potent response towards phagocytosis in carbon clearence
assay. This study demonstrates potent immunomodulatory activities and therapeutic
potential of Stevia rebaudiana ethanolic leaf extract for the prevention of immunological
disorders.
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In Vitro Regeneration of Chlorophytum Borivilianum
using Different Explants – A Rare Medicinal Herb
Dwivedi Rashmi
Abstract: Chlorophytum
borivilianum Sant et Fernard, commonly known as safed musli belonging to the Lilliaceae
family is an important medicinal herb of commercial importance. Chlorophytum borivilianum
with about 17 species found in India, is a very popular herb in traditional Indian
medicine and constitutes a group of herbs used as ‘Rasayan’ or adaptogen. In the
present study, shoot growth and multiplication responses using different explants
were compared on solid medium. Crown and fruit were used as an explant and cultured
on MS medium with different concentrations of BAP (6-benzyl amino purine) at 1.0,
2.0, 3.0, 4.0, 5.0 mg/l and IAA (Indole acetic acid) at 1.0 and 1.5 mg/l. Shoot
proliferation was maximum (75%) with 5.0 mg/l BAP when compared with other combinations
of phytohormones. Fruit explants also showed response in the treatment using 2 mg/l
BAP and 5 mg/l BAP without further dedifferentiation. In vitro grown shoot buds
produce good profuse roots with 75% rooting in half MS medium with 500 mg/l activated
charcoal. 90% survival rate of the plantlets was observed during acclimatization.
This protocol can be successfully utilized for the large scale clonal multiplication
and germplasm conservation of rare valuable medicinal herb.
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PCR-Based Differentiation of Phenotypically similar
Gram-Negative Bacilli recovered from CSF Samples
Nagaveni S., Rajeshwari H., Oli Ajay Kumar, Patil S. A. and Kelmani Chandrakanth
R.
Broad-range bacterial PCR combined with DNA sequencing
was used as a means of identifying clinically relevant isolates of gram-negative
bacilli recovered from cerebrospinal fluid (CSF). Genes coding for bacterial 16S
rRNA genes were amplified. The study was designed to compare phenotypic with molecular
identification. These results indicated that preliminary phenotypic testing had
misidentified several isolates due to the morphological changes induced by the antibiotics.
This invites efforts to be focused on the application of PCR with broad-range bacterial
primers to improve the etiologic diagnosis of bacterial infections.
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Efficacy of Probiotic Toothpaste on subjects with
Periodontitis - A Randomised Double-Blind Study
Philip Koshy , Swaminathan Dasan, Muniandy Sekaran and Yaacob Hashim
Abstract: Probiotics
have been successful in the treatment of digestive related diseases. However, its
use to overcome dental disorders is an innovative approach in Oral Biology. Probiotics
has been shown to possess antibacterial activity and it thus can control pathogens
from causing dental diseases like gingivitis. Gingivitis can lead to periodontitis
when not treated and eventually to loss of teeth. Past studies indicate that probiotics
also lowers the pH so that bacterial dental plaque cannot form mature dental plaque
and calculus that can lead to periodontal disease. In this clinical study, Probiotic
toothpaste is assessed in this randomized parallel, double blind study to evaluate
its efficacy, in comparison to a placebo, on patients with chronic gingivitis or
early periodontitis. The toothpaste contains bacteriocin extracted specifically
from lactic acid bacteria found in Probiotics. This randomized, parallel, double
blind study, in which 60 adult patients diagnosed with chronic gingivitis or early
periodontitis, 30 of whom were subjected to the test Probiotic toothpaste and the
remaining 30 used a control toothpaste evaluated the healing of gingival tissues
using clinical indices. This parallel study was carried out over a period of 2 months.
The clinical indices used in this study were Plaque Index (PI), Gingival Index (GI),
Papillary Bleeding Index (PBI) and Stain Index. The results of this study indicated
that the Probiotic toothpaste containing bacteriocin was marginally significant
in its efficacy to control plaque and gingivitis when compared to the placebo toothpaste
and both the test and control toothpastes had no effect on dental staining.
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Spectrometric Analysis of Quinazoline Alkaloids from
Methanolic Leaf Extract of Adhatoda vasica Nees.
Khadiker S. and Tenguria R.K.
Abstract: Methanolic
extract of leaves of Adhatoda vasica Nees. were prepared using Soxhletion method.
Obtained leaf extract were purified with the help of column and thin layer chromatography
followed by acid hydrolysis and methylation. These isolated and purified compounds
were subjected to spectral analysis. On the basis of Mass spectrum, IR spectrum
and NMR spectrum analysis, it may be concluded that leaf extract of Adhatoda vasica
Nees. had the compounds namely Phenoxy Vasicolinone with molecular weight 413 and
Phenyl Adhatodine with molecular weight 397. Both obtained compounds are the quinazoline
alkaloids in which two fused benzene rings are present.
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Genetic Structure of an Endangered Cycas Revoluta
using RAPD Markers
Sarethy Indira P., Gulati Natasha, Bansal Akanksha, Gupta Vibhuti, Malhotra Kunal
and Gabrani Reema
Abstract: Cycads
are one of the most primitive species and represent a key point in evolution between
ferns and angiosperms. Cycas revoluta is an introduced species in India, widely
exploited for its ornamental and medicinal features, facing extensive habitat destruction.
Eight accessions were collected from three regions. These were considered to belong
to three populations assuming each population to be homogeneous. Genetic diversity
within and amongst accessions was studied using fifteen RAPD primers. Multiple bands
of scorable intensity were reproducibly generated from 10 decamers. A total of
51 loci (82.26%) were found to be polymorphic. Co-efficient of gene differentiation
(GST) was 0.4260, indicating that 57.4% of variation existed within populations.
To remove the upward bias of GST, AMOVA analysis was used to confirm that a non-significant
amount of genetic diversity (25%) was attributable to variance between populations
while a larger share (75%) of the diversity occurred within populations. The principal
co-ordinates analysis also confirmed AMOVA results. Since our sampling was from
locations widely separated (2000 km), this level of genetic diversity could be because
of genetic drift or ancestral polymorphisms. Based on our results and the high level
of genetic diversity observed, conservation strategies should be formulated focusing
on collection of samples from as many locations as possible.
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Molecular Identification of ‘Candidatus Phytoplasma
Cynodontis’ associated with Bermuda Grass Disease in Rajasthan, India
Gaur R.K., Raizada Richa and Gupta Vijai K.
Abstract: Bermuda
grass showing symptoms of white leaf disease has been observed in the area of Rajasthan,
India. The diseased plants showed typical white leaf symptoms, proliferation of
auxiliary shoots, bushy growing habit, small leaves and shortened stolons. Using
polymerase chain reaction (PCR) amplification with P1/P6 universal primer followed
by R16F2n/R16R2 primers, all symptomatic plants tested positively; whereas no amplification
product was obtained from non-symptomatic plants. RFLP analysis of PCR products
with HaeIII, RsaI or AluI endonuclease generated fragment profiles that were identical
for all white leaf samples. The phytoplasma 16/23S intergenic region was sequenced
directly with primers R16F2n/R16R2 and compared by BLAST analysis with those of
other phytoplasma rDNA sequences archived in GenBank. The highest sequence homology
(95%) obtained was with that of ‘Candidatus Phytoplasma cynodontis’ (GenBank Accession
No.Y14645), the type member of the BGWL (16SrXIV) group. This is the first molecular
identification of ‘Ca. Phytoplasma cynodontis’ (16SrXIV group) associated with Bermuda
grass white leaf disease in West India (Rajasthan).
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Isolation, Identification and Media Optimization of
Thermostable Alpha Galactosidase producing Geobacillus Sp.
Chavan Sanjay and Sabharwal Sushma
Abstract: Thermophilic
alpha galactosidase producing bacteria have been isolated by enrichment of the soil
samples with soybean meal for 24 hrs at 60 °C. The isolated strain is aerobic and
gram positive growing, optimally at pH 8.0.This bacterial strain was identified
as a Geobacillus sp. by its colony morphology and biochemical tests. The growth
pattern of the bacteria was studied at various stress conditions. The alpha galatosidase
activity was detected in the culture filtrate after 15 hours of incubation period.
The bacterial strain used different carbohydrates as the carbon source; however
the maximum production of alpha galactosidase occurred when 1.5% (w/v) soybean meal
was used. The use of organic nitrogen favored the production of alpha galactosidase
in the isolated strain compared to inorganic nitrogen sources.
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Studies on Establishment of Primary Cell Cultures
from Hepatopancreatic Tissue of Penaeus Vannamei Post Larvae using Different Biological
Fluid Extracts
Rao Jaganmohan P. and Rao Sambasiva K.R.S.
Abstract: A primary
cell culture system was developed for the cells of hepatopancreatic tissue of Penaeus
vannamei post larvae. Minced tissue fragments of hepatopancreas were disaggregated
with 0.1% trypsin. The cells were seeded at different densities in three different
media viz. minimum essential medium (MEM), Hanks balanced salt solution (HBSS) and
L-15 medium. Epitheliod like cells were developed in all media. The cultured conditions
like time of disaggregation, seeding density, temperature, CO2 concentrations
were optimized. The primary cell cultures were also established by supplementing
all the three media with different biological fluid extracts like Fetal bovine serum
(FBS), Prawn muscle extract (PME) and Fish fry extract (FFE).
Maximum number of viable cells were obtained with 30 minutes disaggregation and
when cells were seeded at a density of 106 cells/ml, primary cell cultures were
best established in L-15 medium among the three media used with different biological
fluids like FBS (20%), prawn muscle extract (PME) and fish fry extract (25%). They
were survived for a period of 50 days. Cell cultures were best survived with 5%
CO2 and at 300C temperature.
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Rapid and Reproducible ‘Green’ Synthesis of Silver
Nanoparticles of consistent shape and size using Azadirachta Indica
Anuradha J., Abbasi Tasneem and Abbasi S.A.
Abstract: Rapid
and reproducible biomimetic synthesis of silver nanoparticles was achieved with
the aqueous extract of neem (Azadirachta indica). The extract served as reducing
as well as stabilizing agent in extracellularly synthesizing silver nanoparticles
at ambient temperatures. At stochiometric ratios differing in a fairly wide band,
the nanoparticle formation was complete within 4-6 hours and exhibited characteristic
plasmon vibration peak at 433±3 nm. It is brought out that reproducible synthesis
of nanoparticles of consistent shapes and sizes is achievable in an environment-friendly
manner.
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Full Length cDNA derived Novel Peptides belonging
to Esculentin Family from Skin of Indian Bronzed Frog Hylarana Temporalis
Reshmy Vimaladevi, Kumar Kesavakurup Santhosh and George Sanil
Abstract: Esculentins
constitute the most potent antimicrobial peptides from frog skin. The present study
describes the successful isolation of 3 novel esculentin -2 peptides from the skin
secretion of Indian bronzed frog Hylarana temporalis. The deduced open reading frames
encoding the biosynthetic precursors of each esculentin consisted of 76 amino acid
residues. Precursors of these novel peptides, named esculentin-2TEa, esculentin-2TEb
and esculentin-2TEc, from H.temporalis showed 12 amino acid differences than the
nearest homologous peptide reported so far. This forms the first report of escuelentin
peptides from Hylarana temporalis, endemic to India and Srilanka.
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Arbuscular Mycorrhizal Symbiosis: An Overview
Jaya Kumari T.R.
Abstract: Arbuscular
mycorrhizas formed by obligately symbiotic soil fungi of the Phylum Glomeromycota
occurs in the roots of more than 95% of plants belonging to diverse ecological groups
in natural as well as disturbed/contaminated habitats. They offer multitudes of
benefits to host plants including enhanced nutrient and water uptake, tolerance/
resistance to biotic and abiotic stresses etc. They have recently been identified
as the most viable strategy to remediate environmental degradations and contaminations
emanating from anthropogenic activities. Their taxonomy is undergoing massive
modifications due to molecular methods of elucidation of the phylogenetic relationships.
The phylum is currently known to comprise approximately 200 species distributed
among ten genera. By far the largest AM fungal genus known is Glomus, with more
than 100 species. Reports on the incidence of arbuscular mycorrhizas in crops of
diverse categories add to the growing body of knowledge that they are ubiquitos,
universal and ecologically significant components of the ecosystems.
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