Technical sessions-Abstracts-Poster Session 5-PS-51-PS-58

36th Ann. Meeting of MSI & Nat. Seminar on Fungal Biodiversity & Bioprospecting….. Goa, India Oct., 29-30, 2009

POSTER SESSION 5

PS – 51

Herbal antimicrobials: A biological approach for purification of water

Awadhesh Kumar*, Rohit K. Mishra, Shalu Srivastava, Amit Kumar Tiwari, Kumar Pankaj Pandey, Ahsan Kamran, Rita Gupta*, Ameritesh Chandra Shukla* and Anupam Dikshit
Biological Product Laboratory, Dept. of Botany, University of Allahabad, Allahabad 211 002.
*Department of Horticulture Medicinal and Aromatic Plants, Mizoram University, Aizwal.

Water purification, is the process of removing bacteria, viruses and other biological contaminants from surface water or groundwater to make it safe and potable for human consumption. A large number of Indian populations residing in rural areas are dependent on the supply of untreated water, which is the root cause of their ailments. The 80% population of rural areas deprived of organized system of collecting, treating and supply of drinking water. The dug wells, hand pumps and step wells are the main sources of drinking water in rural areas. Somewhere the water ponds, lakes or rivers are used for domestic purpose. So that 90% of the rural diseases may be attributed to the polluted water. Unlike cities whereas fairly large population is using water filters based on ultraviolet radiation technology, ozonised water and RO system. The rural population is thriving on the contaminated water supply due to lack of financial resources and other pressing essentialities of life. Therefore, it is vital that with increased emphasis on augmenting the source of drinking water, efforts should also be made simultaneously for ensuring its quality. In rural areas the availability of plant materials used in the purification and its acceptability as environmentally safe has to be ensured. Essential oils, extracted from wide variety of plant species like Cinnamomum zeylanicum, Syzygium aromaticum, Eucalyptus globulus, Mentha piperita, Ocimum kilimandcharium, Vicia faba and Parkinsonia aculeata have potent antimicrobial activity for removing, inhibiting or killing the growth of various microorganism in drinking water.

PS – 52

Aeromycometric investigation in industrially polluted environment (ambattur) in Chennai

**S. Bhuvaneswari, N. Jawahar*, S. Vinodh*, S. Hariharan*, Ashok Livingstone Alex* and N. K. Udaya Prakash*

Research and Development, MARINA LABS, 40, Anna Nedum Pathai, Choolaimedu, Chennai 600 094.
*Department of Biotechnology, Veltech Hightech Dr. Rangarajan Dr. Sakunthala Engineering College,
Alamadhi Road, Avadi, Chennai 600 062.
*E-mail: marinalabs@yahoo.co.in

Ambattur Municipality is situated on the western side of Chennai city at a distance of about 20 km on Chennai - Thiruvallur High Road. Ambattur is fast developing as an urban center. Government of Tamil Nadu has established an Industrial Estate in Ambattur which has attracted reputed manufacturers like TVS, DUNLOP, T.I. CYCLES, etc, to set up industries. A number of housing colonies have sprung up in the area. The people are mainly employed in industrial establishments. The Municipality spreads over an area of 40.36 Sq.km., and the population of Ambattur as per 2001 Census is 3,08,764. The area is densely occupied different industries emanating physical and chemical pollutants within the area. People living on this location report suffering due to suffocation and chemical irritants. In this environment a study on presence of Atmospheric fungi was conducted. The study was conducted during the period of July – August. Altogether a total of 20 different spots were selected and atmospheric fungi were studied by exposing Potato Dextrose Agar through Andersen single stage sampler (Aerotech Inc, Arizona, USA). The exposed plates were brought in the laboratory and the growing colonies were analyzed. The isolated colonies are classified under 26 species among which Aspergillus was represented by maximum (14 spp.) number of species followed by Penicillium (5 spp.). The detailed report on this study will be discussed.


PS – 53

Prevalence and seasonal periodicity of airborne fungal spores in indoors and outdoors of a coastal village in Pondicherry region

B. K. Nayak*, R. Anandu, K. Usha and Anima Nanda**
Dept. of Plant Biology and Biotechnology, K. M. Centre for P. G. Studies (Autonomous), Pondicherry 605 008.
**Dept. of Biomedical Engineering, Sathyabama University, Rajiv Gandhi Salai, Chennai 600 119.

Studies on the prevalence of airborne fungal spores with their seasonal periodicity, in the indoors and outdoors of the houses in a coastal village of Pondicherry was carried out by implementing Petri plate sedimentation method from October 2008 to September 2009. Composition and concentration of fungal pores considerably varied from indoors to outdoors as well as from season to season. Outdoor air harbored maximum fungal spores (53%) in comparison to indoor air (47%). Occurrence of fungal species was predominated with more number of propagules during mid winter (December) and early rainy (July) periods in comparison to other months. In qualitative analysis, out of the total species recorded, Aspergillus was found with the highest frequency and had eleven members i.e., A. awamori, A. fumigatus, A. niger, A. flavus, A. flavipes, A. nidulans, A. ochraceous, A. japonicus, A. terreus, A. versicolor and A. wentii, but quantitatively, Penicillium was isolated highest in its contribution to total CFUs followed by Aspergillus. Out of the 33 isolated fungal taxa, Aspergillus fumigatus, A. awamori, A. niger, Rhizopus stolonifer and Alternaria alternata were the predominant aeroallergens, which cause different types of respiratory/lung diseases in atopic human beings. In seasonal periodicity, winter contributed the maximum spore load (41%) followed by rainy (33%), summer was found the least (26%) in harboring the spore mass in the indoors and outdoors in the village environment. Alternaria alternata, which is accounted as a human allergen for sporosis inducer and an agent for hay fever and other pathologies, was also intermittently recorded. In addition to the above aero allergenic fungi, a few plant pathogenic, saprophytic, field and storage fungi were also recorded during the study period.

PS – 54

Photodynamic control of Trichophyton mentagrophytes var. mentagrophytes by acriflavin, an acridine dye

S. Jenefar*, V. Balaji and P. Ebenezer
Centre for Advanced Studies in Botany, University of Madras, Chennai 600 025.
E-mail: jenesfashion@yahoo.co.in

Photodynamic therapy (PDT) is a treatment that involves light to activate a photosensitizing agent (photosensitizer) in the presence of oxygen. The exposure of the photosensitizer to light results in the formation of Reactive Oxygen Species, such as singlet oxygen and free radicals, causing localized photodamage and cell death. Clinically, this reaction is described as cytotoxic and vasculotoxic. The response of Trichophyton mentagrophytes to treatment with acriflavin, a common photodynamic dye has been studied in an attempt to apply the technique of PDT. Acriflavin, an acridine dye was found to have acted as a photosensitizer, exerting a photodynamic effect on the test fungus. Acriflavin was effective in controlling growth of the Trichophyton mentagrophytes in the presence of light and oxygen at a concentration of 100 µg/mL. Radial growth and biomass decreased in the presence of light and dye. Sporulation and spore germination were affected to a considerable extent. The optiumum pH and temperature for photodynamic control was found to be 7 and 35˚C respectively. The overall lipid, protein and sterol metabolism were found to have been affected by the photosensitizer. The syntheisis of protective enzymes, catalase and peroxidase also decreased with an increase in the concentration of the dye. Damage to the fungal membrane was minimal since the photosensitizer is a permeable dye. The response of Trichophyton mentagrophytes to treatment with acriflavin, and their action detected by antifungal assay.

PS – 55

Extraction, estimation and detoxification of aflatoxins present in storage mycoflora of animal feeds

K. P. Kannan*
Department of Biotechnology
Bannari Amman Institute of Technology, Sathyamangalm Erode District.
*E-mail: kp_kannan2001@yahoo.co.in

Aflatoxins are naturally occurring mycotoxins that are produced by many species of Aspergillus, most notably Aspergillus flavus and Aspergillus parasiticus. Aflatoxins are toxic and among the most carcinogenic substances known. After entering the body, aflatoxins are metabolized by the liver to a reactive intermediate, aflatoxin M1, an epoxide. The native habitat of Aspergillus is in soil, decaying vegetation, hay, and grains undergoing microbiological deterioration and it invades all types of organic substrates whenever conditions are favorable for its growth. Favorable conditions include high moisture content (at least 7%) and high temperature. Crops which are frequently affected include spices like chile pepper, black pepper, coriander, turmeric, ginger. At least 13 different types of aflatoxin are produced in nature. Aflatoxin B1 is considered the most toxic and is produced by both Aspergillus flavus and A .parasiticus.Aflatoxin G1 and G2 are produced exclusively by A. parasiticus. While the presence of Aspergillus in food products does not always indicate harmful levels of aflatoxin are also present, it does imply a significant risk in consumption of that product. In the present investigation animal feeds i.e. Cotton seeds, Different refinery cakes (sunflower cakes, groundnut cakes, and sesamum cakes) were collected from the market and were analyzed for the presence of storage mycoflora and the Aflatoxin concentration were employed. The results will be discussed in detail.

PS – 56

Bioadsorption of heavy metals by metal tolerant filamentous fungi

Peralam Yegneswaran Prakash*, Vaibhav Gupta**, Indira Bairy**
*Mycology Laboratory, Department of Microbiology, Kasturba Medical College.
**Division of Biotechnology, Manipal Life Sciences Centre, Manipal University, Karnataka.

The property of certain types of inactive, dead, microbial biomass to bind and concentrate heavy metals from dilute aqueous is referred to as bioremediation. Certain algae, fungi and bacteria have been employed to affect bioremediation in the event of oil spillages, sludge disposal and industrial waste cleanup in a large scale and involve the solid and liquid phase. Contaminated soils with toxic heavy metal pose a ecological threat and use of filamentous fungi for Bioremediation remains a choice owing to its robust nature and a potential of toxic metal recovery. To investigate bioabsorption properties of metal tolerant filamentous fungi isolated from metal contaminated soil sources by atomic absorption spectrophotometer. The study involved 3 phases; (i) Sampling for isolation of fungi from polluted soil sites (ii) Screening the heavy metal tolerance of the fungi to Copper, Lead, Iron and Cobalt using the Agar dilution and Spore germination assay (iii) Investigation of the bioadsorption properties of the metal tolerant fungi by atomic absorption spectrophotometer. Aspergillus niger, Aspergillus terreus, Absidia corymbifera, Paeciliomyces varioti and Trichoderma viridae were isolated from the polluted soil sites. In comparison to the 5 fungi studied; A. terreus was found to be tolerant to a relatively higher amount of Copper, Lead and Iron in the concentration of 35ppm, 25ppm and 200ppm respectively. The bioadsoprtion was influenced by physical parameters like temperature, pH and live versus dead mycelial biomass of Aspergillus terreus. The potential of Aspergillus terreus in bioremediation owing to their bioadsorption properties were found to be encouraging. Further studies by dose modulation effects on the genetic improvement of the strains could yield a promi
sing bioremediation agent to quench the increasing soil heavy metal toxicity making the planet a better place to live.

PS – 57

Growth control of the dermatophyte Trichophyton mentagrophytes var. mentagrophytes by a dual mode of action with Griseofulvin

D. Raynukaa*, V. Balaji V. and P. Ebenezer
Centre for Advanced Studies in Botany, University of Madras, Chennai 600 025.
*E-mail: raynukaa@gmail.com

Photodynamic therapy (PDT) involves light-sensitive photosensitizer, light, and molecular oxygen. After excitation with visible light, highly cytotoxic singlet oxygen and other reactive oxygen species (ROS) are generated by electron transfer. Singlet oxygen is a very reactive molecule that can damage many components of the target cell, resulting in cell death. Singlet oxygen can also destroy the photosensitizing agent itself preventing further action, a process referred to as photobleaching. Griseofulvin, an antifungal drug was found to have acted as a photosensitizer, exerting a photodynamic effect on the test fungus. The response of Trichophyton mentagrophytes to treatment with Griseofulvin, an antifungal drug, has been studied in an attempt to apply the technique of PDT. Griseofulvin was effective in controlling the growth of Trichophyton mentagrophytes in the presence of light and oxygen at a concentration of 100 µg/mL. Radial growth and biomass decreased in the presence of light and dye. Sporulation and spore germination were affected to an appreciable extent. The optiumum pH and temperature for photodynamic action of grieseofulvin were found to be 7 and 35˚C respectively. The overall lipid, protein and sterol metabolism were found to have been affected by the photosensitizer. The syntheisis of protective enzymes, catalase and peroxidase also decreased with an increase in the concentration of the dye. Damage to the fungal membrane appeared to be minimal since the photosensitizer is a permeable dye and the damage was endogenous. The response of Trichophyton mentagrophytes to treatment with grieseofulvin, and their action detected by antifungal assay.

PS – 58

X-ray induced mutation in Ganoderma species for higher anti thrombotic activity

P. Samuel Naveen* and M. Eyini
Research Centre in Botany, Thiagarajar College (Autonomous), Madurai 625009.
*E-mail: jeyini2005@yahoo.co.in

Thrombosis refers to the formation of a blood clot, thrombus within a vein which can partially or completely block the flow of the blood in the vein causing an embolism in the major organs and is life threatening. The mortality and morbidity resulting from thrombosis has been a major and increasing public health problem in India with a prevalence rate of 200 per 100,000 of the population per year. Thrombosis may be attributed to a sedentary lifestyle due to which disruption to blood flow occurs in the lower limbs and to an inappropriate diet that predisposes to acute thrombotic events. A regular diet containing ingredients with proven anti-thrombotic effect might be a convenient and effective way to prevent the disease. The use of Ganoderma lucidum as a dietary supplement has been documented in Chinese pharmacopoeia since approximately second century B.C. and it has been implicated against several diseases and physiological disorders like hypertension, diabetes and aging-related malfunctions. The present study investigated the effect of a physical mutagen, X–ray in increasing the antithrombotic activity of Ganoderma species in vitro. Physical mutation by X–Ray was induced by exposing PDA plates containing 6 days-old cultures of three pure cultures/wild isolates of Ganoderma lucidum (GL1, GL3, and GL5) and one pure culture of G. applanatum (GA2 IARI4655) for 0.8, 1.2, 1.6, 2.0 and 2.4 seconds at the rate of 40 mA / 75 KV from a X-ray machine (100 mA GE, INDIA). Based on mycelial morphology, growth rate and ligninase enzyme plate assays, eighty mutants were selected for the screening experiments. In the preliminary set of experiments, the PD broth aliquots of 14 days- old cultures of the four parent strains and their respective mutants were used to study the anti-thrombotic effect in whole blood. Among the parent strains, GL1 showed a significant increase in clotting time of 28sec after correcting for the dilution factor. The increase in clotting time of the other parent strains ranged between 21 sec (GA2),18.3(GL3) and 12sec (GL5) compared to the time required for clotting to occur under normal conditions (216sec). The mutants 2XLa4, 2.4XLa4, 2.4XLc4, 2.4XLc2, 1.6XLd4, 2.4XLd3, 1.6XAb3 and 2.4XAb1 showed significantly higher anti-thrombotic effect by extending the clotting time by 71 to 200 seconds compared to the parent strains at room temperature. These eight mutant strains were further subjected to coagulability tests, viz. Prothrombin Time (PT) and the Activated Partial Thromboplastin Time (APTT) using citrated plasma. The parent strains showed an increase in Prothrombin time which ranged between 19sec (GA2) and 49 sec (GL1) over that of the control (22sec) while the APTT values shown by the parents were extended by 6sec (GA2) and 29sec (GL3) over the control (25 sec) after allowing the effects of dilution. The pure culture of G. lucidum (GL1- IARI 1995) and its mutants (2XLa4, 2.4XLa4) showed the highest anti-thrombotic activity. PT & APTT duration of G. applanatum was less than that of the G. lucidum strains but its mutant, 2.4XAb1 showed significantly higher PT & APTT values than the latter. The mutants selected after the exposure time of 1.6, 2.4 and 2.0 seconds viz. 2XLc4, 2.4XLc2 (GL3 mutants) and1.6XLd4 (GL5 mutant).showed the maximum prothrombin time. The longest APTT was shown by 2XLc4, 2.4XLc2 (GL3 mutants) and 2.4XAb1, 1.6 XAb3 (GA2 mutants). PT and APTT results were more selective and confirmatory as some of the mutants of GL1 (2XLa4) and GL5 (2.4XLd3) which showed significant results in the screening tests using whole blood were found to be less efficient than their parents. The liquid growth media and the mycelia of the screened mutants thus represent a potential source for new therapeutic agents to treat thrombosis. However further characterization to study molecular level changes in the mutants are needed which may also pave the way for the production of new nutriceutical compounds for the treatment of thrombosis.