An efficient antagonistic strain of Bacillus subtilis, originally isolated from the rhizosphere of established tea bushes, was found to cause structural deformities in six pathogenic fungi under in vitro culture conditions. This effect was attributed to the production of diffusible and volatile antifungal compounds. Out of the selected test fungi four were phytopathogenic, while the remaining two were of clinical importance. The bacterial strain successfully restricted the growth of all test fungi in dual cultures, and induced morphological abnormalities such as mycelial and conidial deviations. The inhibitory effect caused by volatiles was greater than that by diffusible compounds.
The morphological, biochemical, and physiological characteristics of a phosphate solubilizing and antagonistic bacterial strain, designated as B0, isolated from a sub-alpine Himalayan forest site have been described. The isolate is gram negative, rod shaped, 0.8 x 1.6 microm in size, and psychrotrophic in nature that could grow from 0 to 35 degrees C (optimum temp. 25 degrees C). It exhibited tolerance to a wide pH range (3-12; optimum 8.0) and salt concentration up to 4% (w/v). Although it was sensitive to kanamycin, gentamicin, and streptomycin (<10 microg mL(-1)), it showed resistance to higher concentrations of ampicillin, penicillin, and carbenicillin (>1000 microg mL(-1)). The isolate showed maximum similarity with Pseudomonas putida based on 16S rRNA analysis. It solubilized tricalcium phosphate under in vitro conditions. The phosphate solubilization was estimated along a temperature range (4-28 degrees C), and maximum activity (247 microg mL(-1)) was recorded at 21 degrees C after 15 days of incubation. The phosphate solubilizing activity coincided with a concomitant decrease in pH of the medium. The isolate also exhibited antifungal activity against phytopathogenic fungi in Petri dish assays and produced chitinase, ss-l,3-glucanase, salicylic acid, siderophore, and hydrogen cyanide. The plant growth promotion and antifungal properties were demonstrated through a maize-based bioassay under greenhouse conditions. Although the bacterial inoculation was found to result in significant increment in plant biomass, it stimulated bacterial and suppressed fungal counts in the rhizosphere. The present study is important with respect to enumerating microbial diversity of the colder regions as well as understanding the potential biotechnological applications of native microbes.
Pseudomonas corrugata, a soil bacterium originally isolated from a temperate site of Indian Himalayan Region (IHR) is examined for its antagonistic activities against two phytopathogenic fungi, Alternaria alternata and Fusarium oxysporum. Although the bacterium did not show inhibition zones due to production of diffusible antifungal metabolites, a reduction in growth between 58% and 49% in both test fungi, A. alternata and F. oxysporum, was observed in sealed Petri plates after 120 h of incubation due to production of volatile antifungal metabolites. Reduction in biomass of A. alternata (93.8%) and F. oxysporum (76.9%) in Kings B broth was recorded after 48 h of incubation in dual culture. The antagonism was observed to be affected by growth medium, pH and temperature. The reduction in fungal biomass due to antagonism of bacteria was recorded maximum in the middle of the stationary phase after 21 h of inoculation. The production of siderophore, ammonia, lipase and chitinase in growth medium by P. corrugata were considered contributing to the antagonistic activities of the bacterium.
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A total of 246 fungal isolates representing 36 genera and 72 species were isolated from the soil samples collected from Indian Himalayan region. Twenty-one species belonged to the genus Penicillium alone. All the Penicillium species were screened for phosphate solubilizing activity on Pikovskaya agar at 21°C. Eight species of Penicillium, exhibiting formation of halos (zone of solubilization) around the fungal colonies in qualitative plate assays, were selected for quantitative estimations. In quantitative estimations that were conducted upto day 30 (at 3 days interval), seven species of Penicillium brought maximum solubilization after day 15, while P. oxalicum showed maximum solubilization after day 21 of incubation. The increase in solubilization coincided with decrease in pH of the broth. Acid phosphatase activity was 1.5-2.0 times higher in comparison to alkaline phosphatase. Many of these species showed wide range of tolerance for temperature, pH and salt concentration.
Carrier-based preparations of two plant growth-promoting rhizobacteria (PGPR) viz. Bacillus subtilis and Pseudomonas corrugata, developed in five formulations were evaluated for their growth promotion, rhizosphere colonization, and viability under storage. The effect of these formulations as fresh preparations, and after 6 months of storage at 4°C and room temperature, was also determined. The bacterial inoculants in all the formulations were found to enhance the growth parameters of the test plant species; best results were obtained in case of alginate-based formulations. Maximum numbers of inoculated bacteria were recovered from the rhizosphere of alginate-based formulation-treated plants after 6 weeks of growth. Viability of bacterial inoculants was maximal in alginate beads, and alginate beads supplemented with skim milk formulations, after 180 days of storage at 4°C.
Myiasis of different organs has been reported off and on from various regions in the world. We report a human case of external ophthalmomyiasis caused by the larvae of a sheep nasal botfly, Oestrus ovis, for the first time from Meerut city in Western Uttar Pradesh, India. A 25-year-old farmer presented with severe symptoms of conjunctivitis. The larvae, 3 in number, were observed in the bulbar conjunctiva, and following removal the symptoms of eye inflammation improved within a few hours.
The present study aims to investigate the effect of extraction method on the recovery of flavonoid glycosides, antimicrobials and antioxidants from Ginkgo leaves collected from six different locations in Uttarakhand, Indian Himalaya. Four extraction methods, namely maceration, reflux, shaker and soxhlet were considered, where reflux extracts showed higher antimicrobial antioxidant activity and higher content of flavonoid glycosides. The reference standards of Ginkgo flavonoid glycosides (quercetin, kaempferol and isorhamnetin) and crude extracts were tested for their antimicrobial activity against gram positive and gram negative bacteria and fungi following disc diffusion method and minimum inhibitory concentration (MIC). All the test microorganisms were observed to be inhibited significantly by Ginkgo flavonoids in plate based assays. Correlation coefficients exhibited the extent of contribution of flavonoid glycosides in antimicrobial activity and confirmed the reflux method as a potential method for extraction. Moreover, antioxidant activity as measured by DPPH assay was also found to be higher in reflux method. Significant variation (p < 0.05) in the flavonoid glycosides among the locations was also observed and sample collected from GB6 location was found to be the best for quercetin and isorhamnetin, while GB5 for kaempferol. Significant correlation (r < 0.05, r < 0.001) was obtained while developing the relationship between total flavonoid glycosides and antimicrobials. The present study, thus suggests the reflux method of extraction to be the best for maximum recovery of flavonoid glycosides with higher antioxidant and antimicrobial activities from Ginkgo extract.
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