Rhinosporidium seeberi proven as a fungus for the first time after a century since its discovery

The 18S rRNA gene sequencing of a pure microorganism isolated in pure culture from human rhinosporidiosis cases coded UMH.48 and preserved at 4oC, and, the fungal extracts of biopsy from new cases of nasal rhinosporidiosis were done. Both the sequences were compared for the presence any identical regions by BLAST tool. Astonishingly both the sequences showed 100% identity with each other. The sequences were further compared with the sequences present in NCBI database, followed by sequences of specific organisms like Mesomycetozoa sp and Synchytrium sp. Based on the morphological features, life cycle and BLAST analysis the organism UMH.48 was categorized as a Fungus. The sequences of UMH.48 and sequences from the fungus extracts from new tissue biopsies were deposited in Genbank with accession numbers JN807465 and JN807466 respectively. This paper reports the identity of 18S rRNA sequences between the pure, preserved, isolate with those obtained from biopsies of nasal rhinosporidiosis obtained from totally new cases. Our isolate has been tentatively identified as a lower aquatic fungus with 100% alignment with Colletotrichum truncatum and Glomerulla sps and lesser score similarity with Synchytrium minutum. Yet the absence of a perfect sexual phase or any asexual fungal spores, very rare  microscopic morphology, life cycle and remarkable resemblance with members of lower aquatic fungi led us to surmise (also through personal communication with NCBI, Taxonomy expert) that the isolate is a Fungus (unknown) and not an Ascomycete

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

An integrated process for Industrial effluent treatment and Biodiesel production using Microalgae

The present day necessities and requirements have emphasized the need for a renewable and alternate energy source is very high. Besides, the present day energy resources posse potential threat to the environment by emitting Greenhouse gases (GHGs) etc. An integrated process which involves a model of the wastewater High Rate Algal Ponds (HRAPs) near the industries and establishment of Biodiesel plants nearer to these ponds to produce algal biodiesel along with other byproducts is elucidated.  Wastewater HRAPs also help in sequestering the CO2 emitted by the industries and is used by the microalga for their photosynthesis in turn providing oxygen to the bacterial population which accumulates and degrades the toxic compounds present in the industrial effluents. This integrated process involving cheaper treatment of industrial effluents, production of algal biodiesel, accumulation of toxic  compounds, sequestration of CO2 and various other non-fuel applications contribute to an effective energy management system

Characterization of Alcaligenes faecalis GPA-1 producing thermostable extracellular α-amylase

The bacterium coded GPA-1(isolated by Dr V Thankamani in 1990) was characterized by standard methods including microscopy, special stains, biochemical tests and growth on various types of media for systematic identification up to genus level. With 16S rRNA gene sequencing, the isolate was identified as Alcaligenes faecalis and deposited in NCBI with GenBank Accession number HQ 848384. The isolate was screened for the production of enzymes like amylase, protease and carboxy methyl  cellulase (CMC). This isolate showed a clear zone of lysis on starch agar, yellow zone on CMC agar when stained with Congo red and a clear zone of casein hydrolysis in skim milk agar indicating amylase, cellulose and proteolytic activity respectively. Preliminary characterization of extra  cellular amylase was done. The strain was found to be alkalophilic as it grew well in pH 9.0 and 10.0. The optimum temperature and salinity were found to be 37oC and 3% respectively. Growth curve experiments of the organism in nutrient broth containing 1% starch at varying physical and nutritional parameters were done up to 72 hrs, and the samples were also  tested for pH changes, biomass, total protein, reducing sugars and  α-amylase activity. Soluble starch (1%) in standard nutrient broth and pH 8.0, 37oC, shaking at100 rpm and 40-44 hours incubation were found to be the optimum conditions for maximal enzyme production

In Vitro Antioxidant Activity of Flowers and Fruits of Alstonia scholaris

The ethnobotanical and pharmacological evaluation of plant based chemicals have shown rapid strides in the last few decades. Plants have been a rich source of important therapeutic agents and form the basis of herbal systems of medicine, like ayurveda, resulting in the revival of ancient traditions of medicine. The present study was carried out to investigate the anti oxidant potential of the inflorescence and fruits of Alstonia scholaris using an in vitro model system like DPPH assay and Beta carotene Assay. The methanol extract of the flower showed powerful antioxidant activity by DPPH and Beta-carotene assays in comparison with the standard butylated hydroxy toluene (BHT), l- ascorbic acid. For DPPH assay the IC-50 value was also calculated and was found to have a significant correlation between benzene extract of flower and methanol extract of fruits. Overall, the methanol extracts of flower showed higher anti oxidant activity than the fruit. Keywords: Alstonia scholaris ,anti oxidant potential, DPPH, Beta Carotene.

In Vitro Antioxidant Activity of Flowers and Fruits of Alstonia scholaris

The ethnobotanical and pharmacological evaluation of plant based chemicals have shown rapid strides in the last few decades. Plants have been a rich source of important therapeutic agents and form the basis of herbal systems of medicine, like ayurveda, resulting in the revival of ancient traditions of medicine. The present study was carried out to investigate the anti oxidant potential of the inflorescence and fruits of Alstonia scholaris using an in vitro model system like DPPH assay and Beta carotene Assay. The methanol extract of the flower showed powerful antioxidant activity by DPPH and Beta-carotene assays in comparison with the standard butylated hydroxy toluene (BHT), l- ascorbic acid. For DPPH assay the IC-50 value was also calculated and was found to have a significant correlation between benzene extract of flower and methanol extract of fruits. Overall, the methanol extracts of flower showed higher anti oxidant activity than the fruit. Keywords: Alstonia scholaris ,anti oxidant potential, DPPH, Beta Carotene.