Potential of Microbispora sp. V2 as biocontrol agent against Sclerotium rolfsii, the causative agent of southern blight of Zea mays L (Baby corn)–in vitro studies.

The study was undertaken with the aim of exploring novel and beneficial agro activities of rare actinomycetes like Microbispora sp. V2. The antagonistic activity of Microbispora sp. V2 was evaluated as a biocontrol agents against Sclerotium rolfsii, a soil-borne fungal plant pathogen. The methodology performed for evaluation of biocontrol agent was in vitro evaluation assay which comprised of three tests viz., cellophane overlay technique, seed germination test and Thiram (fungicide) tolerance of Microbispora sp. V2. The isolate was found to inhibit the fungal pathogen Sclerotium rolfsii to 91.43% in cellophane assay. In seed germination assay, Microbispora sp. V2 treated seeds resulted in 25.75% increased germination efficiency, as compared to seeds infected by Sclerotium rolfsii. The isolate Microbispora sp. V2 could tolerate 1000 µg mL-1 of Thiram (fungicide). The in vitro assay studies proved that Microbispora sp. V2 can be used as antifungal antagonist and thus posses’ great potential as biocontrol agent against southern blight caused by Sclerotium rolfsii in Zea mays L (Baby corn) which causes large economical losses.

Heat stable antimicrobial activity of Allium ascalonicum against bacteria and fungi.

To study antimicrobial activity of shallot in comparison with that of garlic and onion against 23 strains of fungi and bacteria, water extracts of garlic, shallot and onion bulbs were prepared. Each extract was studied in different forms for their antimicrobial activity viz., fresh extract, dry extract and autoclaved extract. Minimal inhibitory concentration and minimal lethal concentrations of these extracts were determined against all organisms by broth dilution susceptibility test. Fresh extract of garlic showed greater antimicrobial activity as compared to similar extracts of onion and shallot. However, dried and autoclaved extracts of shallot showed more activity than similar extracts of onion and garlic. Fungi were more sensitive to shallot extract than bacteria. Amongst bacteria, B. cereus was most sensitive (MIC=5 mg ml(-1)). The lowest minimum bactericidal concentration of shallot extract amongst bacteria tested was 5 mg ml(-1) for B. cereus. Amongst fungi, Aureobasidium pullulans and Microsporum gypseum were most sensitive (MIC= 0.15 mg ml(-1)). The lowest minimum lethal concentration was 2.5 mg ml(-1) for Microsporum gypseum and Trichophyton mentagrophytes. It was therefore, expected that the antimicrobial principle of shallot was different than the antimicrobial compounds of onion and garlic. In addition, the antimicrobial component of the shallot extract was stable at 121 degrees C.

A non-polyene antifungal antibiotic from Streptomyces albidoflavus PU 23.

In all 312 actinomycete strains were isolated from water and soil samples from different regions. All these isolates were purified and screened for their antifungal activity against pathogenic fungi. Out of these, 22% of the isolates exhibited activity against fungi. One promising strain, Streptomyces albidoflavus PU 23 with strong antifungal activity against pathogenic fungi was selected for further studies. Antibiotic was extracted and purified from the isolate. Aspergillus spp. was most sensitive to the antibiotic followed by other molds and yeasts. The antibiotic was stable at different temperatures and pH tested and there was no significant loss of the antifungal activity after treatment with various detergents and enzymes. Synergistic effect was observed when the antibiotic was used in combination with hamycin. The antibiotic was fairly stable for a period of 12 months at 4 degree C. The mode of action of the antibiotic seems to be by binding to the ergosterol present in the fungal cell membrane resulting in the leakage of intracellular material and eventually death of the cell. The structure of the antibiotic was determined by elemental analysis and by ultraviolet (UV), Fourier transform infrared (FTIR), nuclear magnetic resonance (NMR) and liquid chromatography mass spectra (LCMS). The antibiotic was found to be a straight chain polyhydroxy, polyether, non-proteinic compound with a single double bond, indicating a nonpolyene antifungal antibiotic.

Production of a growth dependent metabolite active against dermatophytes by Streptomyces rochei AK 39.

BACKGROUND & OBJECTIVE: Dermatophytes responsible for causing dermatophytoses in humans have acquired resistance to certain antimycotic drugs. We isolated naturally occurring actinomycetes with an ability to produce metabolites having antimycotic property. The timecourse of antifungal metabolite production in terms of arbitrary units (AU) under optimum conditions was studied. METHODS: Water and soil samples were collected from various locations. The actinomycetes were isolated on starch casein medium and screened for their antifungal activity against yeasts and molds including dermatophytes. One promising isolate which showed a unique, stable and interesting property of inhibiting only dermatophytes was selected and characterized. Optimization of antifungal metabolite production in terms of AU using Trichphyton rubrum as target was done. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values of the culture supernatant from the isolate and that of griseofulvin were determined for all dermatophytes. RESULTS: Of the 218 actinomycete isolates, 14 per cent produced the metabolites having antifungal activity. The selected actinomycete, identified as Streptomyces rochei AK 39 produced metabolite, which was active against only dermatophytes whereas yeasts and other molds were resistant to it. Starch casein medium was found to be good for inducing antifungal activity in the isolate. The maximum antifungal metabolite production (400 AU/ml) was achieved in the late log phase, which remained constant during the stationery phase, and it was extracellular in nature. The MIC and MFC values of the culture supernatant from the isolate against the dermatophytes were within the range 1.25 to 5 and 1.25 to 10 AU/ml respectively. INTERPRETATION & CONCLUSION: The metabolite from Streptomyces rochei AK 39 was produced during late log phase and was active against only dermatophytes with a greater potency than griseofulvin. However, this needs further investigation using purified powdered form of the active component.

Antimicrobial susceptibility of thermophilic Campylo-bacter spp. isolated from environmental samples.

Environmental samples were subjected to determine frequency of occurrence of pathogenic campylobacters in the environment. The antimicrobial susceptibility of the isolates was tested to evaluate the level of antibiotic sensitive campylobacters in the environment of investigation. In all, 70 Campylobacter isolates were obtained from water and domestic animal faeces samples using Kapadnis-Baseri device and antimicrobial susceptibility of them was determined by disc diffusion test and E- test. The results indicated that all the isolates of Campylobacter were sensitive to ciprofloxacin and resistant to cefotaxime, cephalexin and ampicillin. Lowest MIC values were observed for ciprofloxacin and gentamicin (2 microg/mL) and highest MIC values for ampicillin and chloramphinicol (256 microg/mL). In general, pathogenic Campylobacter spp. were prevalent in large numbers in the environment, however, they were sensitive to ciprofloxacin.

Isolation, characterization and optimization of antifungal activity of an actinomycete of soil origin.

About 312 actinomycetes were isolated from soil samples on chitin agar. All these isolates were purified and screened for their antifungal activity against pathogenic fungi. Out of these, 22% of the isolates exhibited activity against fungi. One promising isolate with strong antifungal activity against pathogenic fungi was selected for further studies. This isolate was from Pune, and was active against both yeasts and molds. Various fermentation parameters were optimized. Based on morphological and biochemical parameters, the isolate was identified as Streptomyces. The correlation of antifungal activity with growth indicated growth dependent production of antimetabolite. Maximum antifungal metabolite production (600 units/ml) was achieved in the late log phase, which remained constant during stationery phase, and it was extracellular in nature.

Effect of gamma radiation on survival of campylobacters in various food samples.

PURPOSE: Campylobacter spp. is a major food borne pathogen and shows resistance towards gamma radiation. In the present study, effect of gamma radiation was assessed on the indigenous strains of Campylobacter spp. inoculated in food and water samples. METHODS: Campylobacter spp. were isolated from river water and faeces of various birds and animals. The growth rate was studied for these isolates by propagating them in Kapadnis-Baseri medium. The survival of Campylobacter spp. inoculated in food and water samples was tested after exposing them to gamma radiation. RESULTS: The isolates survived well in meat and milk samples and were sensitive to 1.8 KGy dose of gamma radiation, which lies with in the FDA limit. The effect of radiation on Campylobacter spp. varied with the species and the type of food. CONCLUSIONS: The results obtained suggest that the dose of gamma radiation should be standardized depending on the Campylobacter spp. and the type of food that is being processed.

Chitin degrading potential of bacteria from extreme and moderate environment.

Five hundred chitin-degrading bacteria were isolated from 20 different locations. High percentage of potent chitin-degraders was obtained from polluted regions. Potent chitin-degrading bacteria were selected by primary and secondary screening. Among the selected isolates, 78% were represented by the genus Streptomyces. Majority of the isolates had good chitinolysis relative to the growth although isolates with better growth were also seen. Such isolates are important for the production of SCP from chitinous wastes. The potent isolates belonged to the genera Streptomyces, Kitasatosporia, Saccharopolyspora, Nocardioides, Nocardiopsis, Herbidospora, Micromonospora, Microbispora, Actinoplanes, Serratia, Bacillus and Pseudomonas. This study forms a comprehensive base for the study of diversity of chitinolytic systems of bacteria.