Technical sessions-Abstracts-Poster Session 3-PS-31-PS-40

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

POSTER SESSION 3

PS – 31

Exploration of starch loaded habitats & samples for isolation of industrially useful amylolytic yeasts

M. Albuquerque and Nandkumar Kamat
Department of Botany, Goa University, Goa 403 206.

Yeasts represent a group of microbes widely distributed in natural ecosystems & significant for their role in food fermentations & enzyme industry. There is relatively scanty knowledge of tropical yeast diversity in natural & fermented media.
Amylolytic yeasts produce amylases that find application in the clarification of fruit juices, stain remover in detergents, preparation of glucose syrup & monosodium glutamate in food processing units, clarification of beer in breweries, as digestive tablets, prevents layering of chocolate syrup & in Desizing textiles.
The screening survey constitutes the first approach in selecting yeast strains exploitable as enzyme producers from various habitats such as tuber cultivating farms, plant storage organs e.g. Potato, sweet potato, cassava.; flour deposits from flour mills, local bread sellers, traditional bakeries. All these habitats can harbor potentially wild amylolytic yeast strains.
Samples were collected from starch loaded habitats in North & South Goa, documented, transported & processed, resulting in the isolation of 119 yeast isolates, which after enzymological screening 15 hyper amylase producers were obtained, which will form a culture bank of wild type amylolytic strains that would be of tremendous use to the food industry.

PS – 32

Bioefficacy of Lactic acid bacteria in the management of Aspergillus japonicus and Rhizopus stolonifer rot of Apricots (Prunus armeniaca L.)

J. Bhadwal and Y.P. Sharma*
Department of Botany, University of Jammu, Jammu, India 180 006.
*E-mail: yashdbm3@yahoo.co.in

Aspergillus japonicus and Rhizopus stolonifer are the most notorious mycopathogens of fresh apricots inciting serious decay and rots. Keeping this in view, an attempt was made to control these two rots using Lactic acid bacteria, an ecofriendly and biosafe biocontrol agent. In the present investigation, lyophilized cells of Lactic acid bacteria were obtained, rehydrated and used in three varying concentrations (1x1012 cfu/ml, 1x1010 cfu/ml and 1x109 cfu/ml) for both pre- and post-infection treatments. It was found that all the three concentrations of Lactic acid bacteria that were used in both pre- and post-infection treatments of fresh apricots inoculated with A. japonicus showed a rot control percentage ranging from 68.81% ± 0.87 to 97.04% ± 0.57 and 37% ± 0.94 to 69.49% ± 1.59 respectively. On the other hand, pre- and post-infection treatments of apricot fruits inoculated with R. stolonifer gave a rot control percentage in the range of 65.18%
± 0.76 to 93.87% ± 0.88 and 0% respectively, indicating clearly relatively more efficacy of pre-infection treatment than post-infection treatment and positive correlation between the concentration of the biocontrol agent and the percent control of fruit rot. It was, thus, concluded that LAB were efficacious and could be used for increasing the shelf-life of apricot fruits.

PS – 33

Phytochemical composition and in vitro antifungal activity of Citrus limon

Mala Majumdar, C. G. Sangeetha, S. Karthik, B. Seema Pai , Krupa B. Paleja and Shwetha H. Gowda
Department of Biotechnology, Centre for Post-Graduate Studies, Jain University,
#18/3, 9th Main, III Block, Jayanagar, Bangalore 560 011.

Citrus limon (lemon) has natural plant chemicals (phytochemicals) which has medicinal value. Lemon also shows some antimicrobial activity. To evaluate the scientific basis of the plant, the pulp, leaves and rind extracts were evaluated against Candida albicans, Aspergillus niger, Fusarium sp .and Penicillium sp. The antifungal activity of the concentrated extracts was evaluated by determination of the diameter of the zone of inhibition against fungi using agar well diffusion method. Aqueous extract of pulp, leaves and rind showed inhibition against C. albicans whereas ethanol extract of pulp inhibited the growth of Fusarium sp .and Penicillium sp. Standard procedures were used to identify the bioactive principles like alkaloids, tannins, saponins, terpenoids, flavonoids and cardiac glycoside. Qualitative analysis revealed the presence of saponins, terpenoids and cardiac glycosides in pulp and tannins, terpenoids, flavonoids and cardiac glycosides in lemon rind. Lemon pulp showed the presence of tannins, terpenoids and cardiac glycosides whereas rind showed the presence of all the phytochemicals except alkaloids and tannins. Quantitative estimation of crude phytochemical constituents showed highest percentage of flavonoids in pulp and saponins in pulp and rind. Antifungal activity of the phytochemical tested showed that flavonoids extracted from pulp showed inhibition against C. albicans whereas saponins from rind and pulp also inhibited C. albicans.

PS – 34

Evaluation of phytochemical composition and antifungal activity of Aegle marmelos

Mala Majumdar, C. G. Sangeetha, Krupa B. Paleja, Shwetha H. Gowda, S. Karthik and B. Seema Pai
Department of Biotechnology, Centre for Post-Graduate Studies, Jain University,
#18/3, 9th Main, III Block, Jayanagar, Bangalore 560 011.

An assessment of phytochemical composition and antifungal activity of aqueous and ethanol extract of bark, leaves, pulp and rind of Aegle marmelos (Bael) against Candida albicans, Aspergillus niger, Fusarium sp .and Penicillium sp. was carried out. The antifungal activities of extracts were compared with standard antibiotics. The antifungal activity of both the extracts was performed by agar well diffusion method. Maximum inhibition of A. niger was recorded by aqueous extract of the leaves and pulp showed inhibition against C. albicans and rind extract showed inhibition against all the tested fungi. The ethanol extract did not show any inhibition against any of the tested fungi. Phytochemical tests were carried out on the aqueous extract using standard procedures to identify the bioactive principles like alkaloids, tannins, saponins, terpenoids, flavonoids and cardiac glycoside. Pulp showed the presence of all the above constituents whereas except tannins all the other active principles were present in bark. Flavonoids and cardiac glycoside were not present in leaves and rind did not show the presence of alkaloid. Quantitative estimation of the percentage of crude phytochemical constituents showed highest percentage of alkaloid in bark and flavonoid in rind. Antifungal activity of the phytochemicals tested showed that the flavanoids extracted from pulp and rind showed inhibition against Candida albicans. Alkaloids and flavonoids from bark extract, saponins from rind extract and alkaloid from bark extract showed inhibition against Penicillium sp. The study suggests that the rind and bark extracts has active principles which could be harnessed as an effective phytomedicine

PS – 35

Biological control of post harvest diseases of brinjal

Reeny Mary Zacharia
Rice Research Station, Moncompu, Alappuzha district, Kerala.

The fungi associated with the spoilage of brinjal fruits were studied. Aspergillus niger and Trichothecium roseum were occasionally present, while fungi like Alternaria solani, Fusarium solani, Colletotrichum gloeosporioides, Botrytis cinerea, Penicillium sp., Rhizopus niger, Curvularia lunata and Botryodiplodia theobromae caused damage through the year. Pythium vexans, Phytophthora palmivora, Pythium aphanidermatum and Rhizoctonia solani were seasonal in appearance and confined to rainy periods. 26 phylloplane fungi were isolated from the leaf surface of brinjal plants. The type of mycoparasitsm between the pathogen and the phylloplane fungi was studied. The pathogens viz., F. solani, A. solani were overgrown by the phylloplane fungi viz., T.viride, A.niger, and A.flavus. Homogenous intermingling was noticed with Phoma sp. and A. solani. Cessation of growth at line of growth was shown by P.palmivora with both pathogen, while a clear zone of inhibition was noticed with F. solani and A.terreus. The mode of action was studied for T.viride alone. Penetration and coiling of hyphae was noticed with F. solani, while penetration and disintegration was common with A. solani. In Trichoderma treated fruits eventhough symptoms were visible by fourth day, it caused only 48 percent damage by 12th day, while in control samples of F.solani infected fruits symptoms were observed by 12th day and complete rotting by 12th day. The study thus highlighted the possibility of utilizing Trichoderma as a biocontrol agent against the major diseases of brinjal.

PS – 36

Occurrence and distribution of keratinophilic fungi in soils of Khardung La, Ladakh (India)

Sandeep Kotwal* and Geeta Sumbali
Department of Botany, University of Jammu, Jammu 180 006.
*E-mail: s.kotwal@yahoo.com

Keratinophilic fungi are an ecologically important group of microorganisms, which cycle keratin that is one of the most abundant and highly stable animal proteins on the earth. This unique group of fungi consists of vigorous and self- sufficient saprophytes as long as environmental conditions are favourable. However, they are opportunists and may become parasitic by accident. After becoming pathogenic, they are able to survive and multiply at host’s body temperature, causing further infection by invading fresh keratinized tissues. It is the only type of fungal infection that is known to be of contagious type.
Realizing the significance of this unique group of fungi, research work was undertaken to study the occurrence and distribution of keratinophilic fungi from the unexplored soils of Khardung La (Ladakh), which is the highest motorable pass of India. It is about 17,582 feet above sea-level, geographically diverse region with cold climate, thick snow cover and comparatively less activity of humans and animals. Soil samples were collected from this area and keratinophiles were recovered by using keratin-bait technique.
Examination of the soil samples showed that 80% of them were positive for keratinophilic fungi. A total of seven keratinophilic fungal species were recovered and all of them produced only the imperfect state on Sabouraud’s dextrose agar (SDA) medium. The recovered fungal species consisted of Aspergillus flavus, A. sydowii, A. ustus, Beauveria bassiana, Fusarium oxysporum, F. sporotrichioides and F. moniliforme var. subglutinans. Among these, Aspergillus flavus, Beauveria bassiana and Fusarium oxysporum are known to be potential pathogenic fungi. During the study, Beauveria bassiana was found to be the most dominant keratinophilic species with 40% frequency occurrence; followed in decreasing order by Aspergillus flavus (30%), Fusarium oxysporum (30%), A. sydowii (20%), A. ustus (20%), F. sporotrichioides (20%) and F. moniliforme var. subglutinans (10%).
Low occurrence and distribution of keratinophilic species from Khardung La soils clearly indicates that harsh environmental conditions and meagre presence of keratinous substrates affect the growth, development and diffusion of keratinophilic fungi.

PS – 37

Study on the efficacy of the vegetative and spore inoculum of Pisolithus albus on the Casurina junghiniana

R. Shenbhagaraman* and V.Kaviyarasan
Centre for advanced studies in Botany, University of Madras, Guindy campus Chennai 600 025.
*E-mail: shenbhagaraman@rediffmail.com, shenbhagaraman@yahoo.co.in

The fruiting bodies of ectomycorrhizal fungus Pisolithus albus were collected from Eucalyptus plantations in around Chennai. The fungus was isolated and maintained over MMN medium. The mycelial growth was optimized and grown over the sterile vermiculite in order to use as vegetative inoculum. Spores were collected from matured fruit bodies were used as spore inoculum. The vegetative inoculum and spore inoculum were applied to one month old seedlings of Casurina junghuniana grown in sterile sand: red soil mixer in the ratio of 2: 1. The seedlings were maintained in the glass house for a period of one month and then transferred to the field. Ten seedlings were sacrificed from each group at the end of the fourth, six, eighth month and were studied for biomass, shoot height, root height, percentage of mycorrhization, starch, total chlorophyll, total carbohydrates, total protein, total nitrogen and micronutrients. The vegetative inoculum enhanced more growth than the spore inoculum. However the seedlings treated with spore inoculum is better than control. The results are discussed in detail.


PS – 38

Studies on characterisation of mycotoxigenic fungi from oil seed cakes and animal feeds

G. L. Sujatha and Ramalingappa
Department of studies in Microbiology, Kuvempu University, Shivagangothri Campus,
Davangere 577 002, Karnataka.

Mycotoxigenic fungi are the potent contaminants of storage foods and feed. The major contaminants are Aspergillus, Penicillium, Fusarium, most species of Aspergillus and Penicillum are able to grow on a wide range of organic substrate. Most of the feed and or feed ingredients are tested for the presence of toxigenic fungi in India. The oil seed Cakes and animal feed samples from Davangere were mycologically investigated in terms of overall fungal diversity and toxicological potential of isolated fungi. Totally 10 samples, 3 samples are oil seed Cakes and all other are animal feeds were randomly collected at intervals from new packets of feed samples in the market in Davangere and brought to laboratory and samples named as S1, S2, S3, S4, S5, S6, S7, S8, S9 and S10 respectively. From these samples mycotoxigenic fungi were isolated by Serial dilution method and direct plate method using different mycological media. Totally 13 genera of fungi was isolated. Aspergillus and Penicillium was identified upto species level. A.flavus detected rapidly on Aspergillus differential medium (ADM) and its toxigenicity tested by ammonia Vapour test. For Penicillium which produces cyclopiazonic acid or other alkaloids detected by Ehrlich test. Aflatoxins from A.flavus and A.parasiticus was purified and separated by Analytical Thin layer chromatographic method. Identified as AFB1, AFB2 & AFG and compared with standard AFB2. Cyclopiazonic acid from P. commune was also purified and identified by characteristic bluish-violet Pigment. By this the incidence of contamination of fungi in oil seed Cakes and animal feeds can be determined.

PS – 39

Laboratory Evaluation of the Entomopathogenic fungi, Aspergillus niger and Metarrhizium anisopliae for Controlling Selected pests

K. N. Varalakshmi and B. S. Kumudini
Department of Biotechnology, Centre for Post Graduate Studies, Jain University, #18/3, 9th Main, 3rd Block, Jayanagar, Bangalore 560 011.

Continuous use of chemicals at high rates against key insects led to pest resurgence resulting from development of resistance and destruction of natural enemies. Entomopathogenic fungi which are very virulent have been used in recent days to develop a logical approach. Application of integrated pest management greatly decreases the use of conventional pesticides, while crop yields remain high. In an attempt to identify such entomopathogenic fungi a study was conducted. In this study fungi were isolated from adult cockroaches, house fly, worms from cauliflower and cabbage from different locations. After proper identification of the fungi, infectivity trials were conducted on Sitophilus oryzae (rice weevil) and eggs of Periplanata americana (cockroach). Different fungi that were isolated were identified as Aspergillus niger, A. flavus, Trichoderma sp. and Penicillium sp. Infectivity trials with A. niger spores showed that by day 8 percentage mortality was 96+0.4 whereas, in control boxes percentage mortality was only 10+0.3. Results of the experiment showing the hatching rate (%) of P. americana eggs sprayed with A. niger spores was very encouraging. In the experimental boxes the percent hatching was greatly affected by A. niger infection. By 7 weeks after infection with A. niger spores, the hating rate was only 6.7% whereas in the control eggs of P. americana the hatching rate is 66.7%. Results of experiment showing mortality (%) of P. americana young ones sprayed with Metarrhizium anisopliae spores showed 100% mortality by the end of day 9. Further field infectivity trials will be carried out for evaluating the feasibility of using these fungi for the biological control of pests of economic importance.

PS – 40

Keratinophilic fungi of domestic animals at Gulbarga Region

G. M. Vidyasagar* and Narayan Hosmani
Medicinal Plants and Microbiology Laboratory, Department of P G Studies and Research in Botany, Gulbarga University, Gulbarga 585 106, Karnataka.
*E-mail:gmvidyasagar@rediffmail.com

Domestic animals play an important role in the transmission of dermatophytes and other potentially pathogenic fungi. Cow, ox, buffalo, sheep and goat are the common domestic animals in Gulbarga region and are very close to human life. Fifty hair samples from each animal were collected in sterilized polythene bags from different places of Gulbarga and incubated them on moistened sterile soil for six weeks at room temperature. The fungi grown on hair samples were transferred on to Sabouraud’s dextrose agar medium and identified with the help of literature. A total of 32 fungal species were identified. Of which, 10 were dermatophytes and 22 were other related ones. Ox hairs yielded maximum of 21 keratinophilic fungi, followed by Cow (20), goat (18), sheep (17) and buffalo (16). Maximum numbers of dermatophytes were isolated from cow and ox hairs (7). This was followed by buffalo (5), goat (4) and sheep hairs (3). Microsporium gypseum was recorded on ox, buffalo and cow hairs, while Trichophyton mentagrophytes recorded only on cow and ox hairs. No incidence of these two dermatophytes was observed on sheep and goat hairs. The occurrence of dermatophytes correlates with the relation of man with animals.