Screening and Characterization of Calcite Solubilizing Bacteria isolated from Marble Slurry dumped in Sukher, Udaipur, Rajasthan, India

Rajasthan is known as mineral majestic state as more than 50 types of minerals are produced including marble. Various types of marbles are spread over in 16 belts in 33 districts of the State. Large amount of marble slurry is discharged as a waste generated by the quarries which is dumped in empty pits in the forest area; thereby creating huge dumping area. The present study was undertaken for screening and characterization of calcite solubilizing bacteria from marble slurry collected from dumping yard in Sukher, Udaipur, India. Screening of calcite solubilizing bacteria was done on calcite agar medium. Calcite solubilizing efficiency (CSE) and calcite solubilizing activity index (CSAI) of the isolates were determined using 0.1% calcite agar medium at 37C for 10 days of incubation. Characterization of the isolates was done on the basis of morphological, biochemical and molecular methods. A total of 27 isolates appeared on calcite agar medium after an incubation of 24 hours at 37°C. Among them only 3 isolates namely isolate CS6, CS13 and CS25 showed clear halo zone of 20 mm, 29 mm and 2mm diameters respectively after 10 days of incubation indicating demonstrable calcite solubilizing activity. The maximum CSE and CSAI 362% and 4.62 respectively were observed for isolate CS13. Isolates CS6, CS13 and CS25 were identified as Exiguobacterium aquaticum CS6 (accession no. MK353511), Staphylococcous aureus CS13 (accession no. KY694446) and Bacillus endophyticus CS25 (accession no. MK353513) respectively. Two isolates that showed remarkable calcite solubilizing activity can be further analyzed for their possible use in restoration of marble slurry contaminated soil.

African histoplasmosis of the penis

African Histoplasmosis is deep mycosis caused by Histoplasma duboisii and genitourinary involvement is extremely rare. We report a case of African histoplasmosis in a 27-year-old subject with painful penis ulcer. Ulcer edge biopsy had revealed inflammatory granulomas made of epithelioid cells, lymphoplasmocytes, polynuclear eosinophils and giant multinucleated cells, with ovoid yeasts surrounded by a clear halo. PAS and Grocott stains revealed numerous fungal structures with a morphology measuring 7 to 15 nm. The diagnosis Histoplasma capsulatum var. duboisii was placed and the patient put on itraconazole (400 mg/day) for six months with a good course. African histoplasmosis of the subject penis is an extremely rare entity. The diagnosis of certainty often makes use of histology and mycological examination, and makes it possible to eliminate differential diagnoses such as cryptoccocosis, tuberculosis or cancer.

Comparative Evaluation on Tannase Production by Lasiodiplodia plurivora ACN-10 under Submerged Fermentation (SmF) and Solid State Fermentation (SSF)

Aim: Tannase (tannin acyl hydrolase, E.C. 3.1.1.20) catalyzes the hydrolysis of ester bonds from complex hydrolysable tannins with the production of gallic acid and glucose and possess broad applications in biotechnology. This study is aimed at the production of tannase by Lasiodiplodia plurivora ACN-10 in SmF and SSF using Terminalia cattapa (almond leaves) and Magnifera indica (mango leaves) as substrates. Study Design: The design adopted to evaluate the production of tannase is submerged (SmF) and solid state (SSF) fermentation. Study Area: Federal Institute of Industrial Research Oshodi, Lagos State Nigeria. Methodology: Fifteen different soil samples were indiscriminately collected within Oshodi, Lagos, Nigeria and were inoculated on PDA plates at 30°C for 3-5 days. A total of 30 isolates was screened on Czapek dox minimal agar incorporated with 1% tannic acid and plates were incubated for 96 h at 30°C. Fungal isolates which were able to disintegrate tannic acid produced a clear halo zone around the colony diameter and were selected to be positive for tannase activity. The best isolate was identified based on its morphological, microscopic and molecular characteristics. Thereafter production and extraction of tannase was carried out in SmF for 0-120 h and in SSF for 0-144 h using Terminalia cattapa (Almond leaves) and Magnifera indica (Mango leaves) as substrates. Results: The total fungal count ranged from 1.0×104 to 3.5×105 CFU/g. A total of 30 fungal isolates produced clear halo zones (ranging from 20 to 70 mm) around the colonies during the screening with tannic acid. Isolate ACN-10, which showed the highest tannic degradation was identified based on its morphological and microscopic characteristics. On the basis of 18S rRNA gene sequence studies, the isolate was identified as Lasiodiplodia plurivora strain ACN-10 and the sequence was submitted to the Genbank with the accession number: MG250374. Results obtained in this study indicated that both substrates can be used by the isolate for tannase production in both SmF and SSF. The result of our investigation on the use of Terminalia cattapa (almond leaves) and Magnifera indica (mango leaves) as substrates for tannase production showed that optimum yield (6.064 U/ml) was obtained at 120 h in SSF while optimum production (4.623 U/ml) was observed in SmF at 96 h using Terminalia cattapa as substrate. Conclusion: Results obtained from this study indicated higher tannase production in solid-state fermentation compared to submerged fermentation. This is the first report to the best of our knowledge that Lasiodiplodia plurivora strain is implicated in tannase secretion. The result also demonstrated high production of extracellular tannases from low cost substrates which can be optimized and scaled up for industrial processes.

Characteristics of Inorganic Phosphate-Solubilizing Bacteria from the Sediments of a Eutrophic Lake

Inorganic phosphate-solubilizing bacteria (IPB) are an important component of microbial populations in lake sediments. The phosphate that they decompose and release becomes an important source of phosphorus for eutrophic algae. The IPB strains were screened and isolated from the sediments of Sancha Lake using National Botanical Research Institute’s phosphate (NBRIP) plates. Their taxonomy was further determined by the 16S rDNA technique. The tricalcium phosphate-solubilizing ability of obtained IPB strains was evaluated using NBRIP- bromophenol blue (BPB) plates and Pikovskaya (PVK) liquid medium. Then, the ability of IPB strains to release phosphorus from the sediments were investigated by mimicking the lake environment. In this study, a total of 43 IPB strains were screened and isolated from the sediments of Sancha Lake, belonging to three phyla, eight families, and ten genera. Among them, two potentially new strains, SWSI1728 and SWSI1734, belonged to genus Bacillus, and a potentially new strain, SWSI1719, belonged to family Micromonosporaceae. Overall, the IBP strains were highly diverse and Bacillus and Paenibacillus were the dominant genera. In the tricalcium phosphate-solubilizing experiment, only 30 of the 43 IPB strains exhibited clear halo zones on plates, while in the liquid culture experiment, all strains were able to dissolve tricalcium phosphate. The phosphate-solubilizing abilities of the strains varied significantly, and the strain SWSI1725 of the Bacillus genus showed the strongest ability with a phosphate-solubilizing content of 103.57 mg/L. The sterilized systems demonstrated significantly elevated phosphorus hydrochloride (HCl–P) decomposition and release from the sediments after the inoculation of IPB strains, whereas no significant effect was demonstrated on the phosphonium hydroxide (NaOH-P). Thus, the IPB strains in the sediments of Sancha Lake possessed rich diversity and the ability to release phosphorus in sediments.

Screening of Cellulase and Pectinase by Using Pseudomonas fluorescence and Bacillus subtilis

A study was conducted to determine the Production of cellulase and pectinase enzyme by using Plant growth promoting rhizobacteria like Pseudomonas fluorescence and Bacillus subtilis. These to micro organism are isolated by serial dilution method. One gram of soil sample was diluted in to 10 ml of sterile distilled water and 1 ml of sample solution was serially diluted in 9ml of sterile water up to 10 dilution. Each sample from dilution 10-5 and 10-6 were taken and streaked in to KB and NA medium and incubate at 24 hrs. After 24 hrs Pseudomonas fluorescence and Bacillus subtilis was observed in the medium of KB and NA medium. Both the culture was sub cultured and maintain in the same for the further work. CMCase medium was prepared and sterilized by autoclave for 121 °C for 15 minutes after sterilization these medium contain petriplate was streaked by bacteria and incubates for 48h after incubation period a clear halo zone was produced by these bacteria among these bacteria Pseudomonas fluorescence are able to produce high amount of cellulose compare to Bacillussubtilis. Pectin agar medium was prepared and sterilized by autoclave for 121 °C for 15 minutes after sterilization these medium contain petriplate was streaked by bacteria incubates for 48h after incubation period a clear halo zone was produced by these bacteria, among these bacteria Pseudomonas fluorescence are able to produce high amount of Pectinase compare to Bacillus subtilis. Plant growth promoting rhizobacteria (PGPR) are beneficial bacteria that colonize plant roots and enhance plant growth by a wide variety of mechanisms.

Validation of RAPID' Staph for Enumeration of Staphylococcus aureus in Foods

RAPID' Staph (Bio-Rad Laboratories, Hercules, CA) is a medium for differentiation and enumeration of coagulase-positive Staphylococcus aureus in food. RAPID'Staph medium is based on a Baird Parker formula optimized for the detection and enumeration of S. aureus in 24 h. The principle of the medium relies on the capacity of S. aureus to reduce tellurite (production of black colonies) and to provoke proteolysis of egg yolk (production of clear halo around the colony). Four foods (pasteurized whole milk, custard pie, processed ham, and smoked salmon) were selected to compare the performance of RAPID' Staph agar to AOAC Official Method 975.55. Method comparison studies demonstrated excellent agreement between the methods. Inclusivity and exclusivity rates of the medium were 100%. RAPID' Staph agar performed as expected when minor procedural variations were introduced, validating the ruggedness of the method. There was no difference in performance between the dehydrated and ready-to-use formulations of the media.

Detection of Early Cytomegalovirus Infection in Pathologial Specimens by H&E, IHC, ISH, Grocott’S and Modified Steiner Silver Stain for LM and EM: A Review

The diagnosis of cytomegalovirus (CMV) in tissue sections is usually quite easy because of the characteristic appearance of intranuclear inclusions. At times the morphological features are equivocal, the nuclear findings may be atypical or only a single cell may be affected, thereby creating uncertainty about the diagnosis. This report reviews our experience with the various microscopic techniques that can be used to make definitive identification of CMV.EM - Visualizing the viral particles in a single or even lysed cell by routine electron microscopy can make the definite diagnosis, but section size limits the usefulness in the detection of early CMV infection (fig. 1). However, if a diagnosis can not be made with certainty by the following methods, EM can be performed on stained paraffin sections for confirmation.H&E - A Hematoxylin & Eosin stain that discloses characteristic intranuclear inclusions surrounded by a clear halo indicates CMV infection (fig.2A).

The core cylinder in the junctional SR of single intact skeletal muscle fibers of the frog after quick-freezing: Its topography

The "core cylinder" (CC) represents confluent electron lucencies seen in the junctional SR (JSR) when skeletal muscle is fixed in the presence of cations such as ruthenium red. CCs are a consistent feature in quick-frozen muscle fibers (Fig.l), both in the resting state and after various intervals between stimulation and freezing, incl. tetanus (5 sec, 50Hz). The CCs are excentrically located inside the JSR and are separated from the junctional membrane toward the transverse tubule by the coextensive line(CL) and from the rest of the membranous envelope of the JSR by granular material (junctional granules, JG). JGs are also found elsewhere in the free SR. Whereas the JGs fill the JSR completely in many cases, often they are separated from the JSR envelope by a peripheral clear halo except in the region of the CL; in favourable sections they form regular rosettes about the CC (Fig.3). Very frequently, the CC loose their more circular configuration (in one dimension)(Fig.4), with flares connecting the CCs with the peripheral halo at either end of the CL (Fig.3). The junction between the electron-lucent flares and the ends of the CL subtends, approximately, the location of the staggered rows of pits on the E face of the freeze-fractured JSR(Fig.2).