UMH.48 (NCBI JN807465) the Fungus causing Rhinosporidiosis is sensitive to anti fungal

UMH.48, JN807465, a fungus causing Rhinosporidiosis was isolated in pure culture from biopsies from patients with nasal Rhinosporidiosis. It was  identified as a lower aquatic fungus by 18S rRNA gene sequencing which compared 100% similar to the sequences from fungal extract of the tissue, thus establishing the etiologic role of UMH.48 in Rhinosporidiosis. UMH.48 18S rRNA sequence showed significant similarity with Synchytridium minutum and very low varying percentages of similarity with Mycobacterium sps., Corynebactrium sps., and Actinomycetales. The organism was tested for susceptibility and sensitivity to antibiotics and antifungal drugs such as  Norfloxacin, Dapsone, Rifampicin and Amphotericin B. UMH.48 was highly sensitive to Amphotericin B and Rifampicin. It was resistant to Dapsone at the concentrations tested

Bacillus isolates VTGP. A-D. 30808 Alcaligenes sp., Exiguobacterium sp., B. pumilus and B. fusiformis producing extracellular alkaline proteases, amylases and cellulases - a preliminary report

Garden soil samples collected from Angamali, Kerala, India were screened for potent bacteria capable of synthesizing extracellular hydrolytic enzymes. Four bacteria were obtained in pure culture. The isolates were systematically identified by microscopy, Gram and special staining techniques for capsule and spores, biochemical reactions and phylogeny by molecular techniques like 16 S rRNA ene sequencing followed by Blast analysis. Production of protease, cellulase and amylase were detected by inoculating nutrient agar containing casein/ skim milkagar, carboxy methyl cellulose and soluble starch respectively.  Alkalophilic and thermophilic properties were investigated by inoculation and incubation of the isolates on specific nutrient media at pH 7-12 and at a wide range of temperatures 28-30, 37, 50 and 650.C. The isolates were coded VTGP. A-D 30808. All the four expressed significant alkalophilic growth at pH 7-12. With respect to protease activity all  except A showed marked protease activity over a high pH range pH 7-12(A-115, B-1119, C-1500, D-1350 Units / ml of liquid culture   supernatant). Both C & D secreted protease as early as 8-12 hours on nutrient agar with 0.1% skim milk forming a clear wide zone of casein hydrolysis. Hence the proteases produced were highly alkalophilic. Amylase activity was marked in all (A-37.38, B-27.58, C-27.92, D-34.82 units per ml culture supernatant). On CMC agar, all the four isolates showed CMCase activity indicated by pale yellow zone of hydrolysis of carboxy methyl cellulose agar when tested with Congo red reagent. A, B and C were strongly positive with minimal visible activity in D. But when tested in CMC broth culture the activities were A-6.71, B-4.30, C-6.56 and D 0.58 units/ ml of culture supernatant). 16S r RNA gene  sequencing of isolates A to D showed maximum alignment with Alcaligenes sp., Exiguobacterium sp., Bacillus pumilus and B. fusiformis. The sequences have been deposited in GenBank with Accession  numbers HQ 848384, HQ 848385, HQ 848386, and HQ 848387

Biological characterization of a fast growing non-sporing alkalophilic lignin degrading fungus MVI.2011

MVI.2011 a rapidly multiplying alkalophilic non sporing fungus was isolated in 1990 and preliminarily identified as a Deuteromycete. The isolate was characterized in detail. The original isolate produced highly fluffy, cottony, fragile aerial mycelia on SDA and a similar growth in liquid SDB also. The organism grew out even on the surface of the conical flask containing the liquid medium inoculated indicating the high aerobic nature. With frequent sub culturing over 20 years the colony morphology on the same media appeared very confined with regular margin and dry surface. Yet there were no reproductive structures. LP staining showed dimorphism with apical fragmentation and no conidia, spores sexual or asexual etc. The pH range was very wide 5-11. The optimum cultural conditions for lignin degradation were pH 8.5, temperature 25-28oC, 12-18 hours and medium- 1% glucose, 0.5% peptone in basal mineral medium. The isolate could breakdown and decolourise commercial lignin (0.1-5%) and alkaline wood extract (1-50%) within 12-18 hours in static cultures evidenced by a clear reduction in absorption at 380 nm (lignin) and a marked shift to increased absorption at 360 nm and between 180 and 300 nm indicating appearance of lignin breakdown products. In optimised  media containing commercial lignin (0.1%) and alkaline wood extract (10%), MVI.2011 secreted Lignin peroxidase (9.39 units/ml), Manganese peroxidase (2.093 units/ml) and laccase (3.5 units/ml) enzymes. The  above data led us to conclude that the isolate was novel being highly alkalophilic, capable of rapid growth, decolourisation of lignins and  secretion of lignin degrading enzymes. Based on microscopic morphology and colony features, the isolate coded MVI.2011 has been identified as “Uncultured Fungus†with NCBI Accession No JN606084. It has been  deduced to be a member of Mycelia sterilia group