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1.
Biodegradation ; 19(5): 739-48, 2008 Sep.
Article in English | MEDLINE | ID: mdl-18288576

ABSTRACT

The untanned proteinaceous tannery solid waste, the animal fleshing (ANFL), was used as substrate in the treatment process (hydrolysis and fermentation) involving Synergistes sp. The nonionic surfactant (Tween 80) was evaluated for its ability to influence on microbial growth and enzyme activity in the hydrolysis and fermentation of ANFL. The addition of Tween 80 in the process significantly increased the activities of hydrolytic and fermentative enzymes like protease (338-360 Um l(-1)) and deaminase (187-206 Um l(-1)) compared to that of control (protease 195-220 Um l(-1) and deaminase 70-83 Um l(-1)). The total viable bacterial count was increased more than twofold, compared to the control in the presence of 0.15% Tween 80. The ANFL fermentation and formation of other metabolites were evidenced by Gas Chromatography and Mass Spectroscopy (GC-MS), Proton Nuclear Magnetic Resonance spectroscopy ((1)H NMR) and Fourier transform infra red spectroscopy (FT-IR). The breakdown of fibrillar proteins in ANFL was confirmed by the scanning electron microscopy (SEM) and the transmission electron microscopy (TEM).


Subject(s)
Fermentation , Industrial Waste , Surface-Active Agents/chemistry , Tanning , Chromatography, High Pressure Liquid , Gas Chromatography-Mass Spectrometry , Hydrolysis , Magnetic Resonance Spectroscopy , Microscopy, Electron , Oxidation-Reduction , Spectroscopy, Fourier Transform Infrared
2.
Bioresour Technol ; 99(7): 2364-72, 2008 May.
Article in English | MEDLINE | ID: mdl-17601727

ABSTRACT

The untanned proteinaceous tannery solid waste, animal fleshing (ANFL), was used as a substrate for acid protease production by Synergistes sp. The strain was isolated from an anaerobic digester used for the treatment of tannery solid waste and was selected for its enhanced protease production at activity 350-420 U/ml. The optimum pH was in the acidic range of 5.5-6.5 and optimum temperature was in mesophilic range of 25-35 degrees C. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the zymogram analyses of the purified protein indicated an estimated molecular mass of 60 kDa. This protease could be classified as aspartic protease based on its inhibition by aspartate type protease inhibitor pepstatin and on non-inhibition by 1,10-phenanthroline, EDTA, EGTA and phenylmethylsulfonyl fluoride. The degradation of ANFL was confirmed by Gas Chromatography-Mass Spectroscopy (GC-MS), Proton Nuclear Magnetic Resonance Spectroscopy (H1 NMR) and Scanning Electron Microscopy (SEM) analyses. In this study we found that the activity of acid protease depended on factors such as calcium concentration, pH and temperature. Based on these lines of evidence, we postulate that this protease is a highly catalytic novel protease of its type.


Subject(s)
Bacteria/metabolism , Fermentation , Tanning , Base Sequence , DNA Primers , Electrophoresis, Polyacrylamide Gel , Gas Chromatography-Mass Spectrometry , Hydrogen-Ion Concentration , Microscopy, Electron, Scanning , Particle Size
3.
J Biomed Mater Res A ; 80(3): 581-91, 2007 Mar 01.
Article in English | MEDLINE | ID: mdl-17031822

ABSTRACT

The silver (0.5-3 at %) substituted nanosize hydroxyapatites (AgHAs) were synthesized by microwave processing. The X-ray diffraction (XRD) peaks are very broad, indicating that the AgHAs were of nanosize (30 nm). Transmission electron microscopy analysis shows needle-like morphology of AgHA, having length 60-70 nm and width 15-20 nm. The AgHA phase was stable up to 700 degrees C without any secondary phases. The antibacterial effect of AgHA against Escherichia coli and Staphylococcus aureus was observed by spread plate method, even for low concentration of silver ions (0.5%) with 1 x 10(5) cells/mL of respective bacterial culture, after a 48 h incubation period. However, some colonies of E. coli were seen with a high dose of 1 x 10(8) cells/mL after 24 h. The zone of inhibition by disc diffusion test method was found to vary with the amount of silver in the sintered AgHA pellets, for both the bacteria, after 24 h of inoculation. Osteoblast cell attachment in varying density was noticed on AgHA samples with 0.5, 1.0, and 1.5% silver substitution. However, osteoblast spreading was significantly greater on 0.5% AgHA compared to 1.0 or 1.5% substituted AgHA samples. Thus, the low amount of AgHA has a potential of minimizing the risk of bacterial contamination, without compromising the bioactivity, and is expected to display greater biological efficacy in terms of osseointegration.


Subject(s)
Anti-Bacterial Agents , Hydroxyapatites , Osseointegration/drug effects , Osteoblasts/cytology , Silver , Tissue Engineering/methods , Bacterial Infections/prevention & control , Biocompatible Materials/chemistry , Biocompatible Materials/pharmacology , Cell Adhesion , Cell Line , Escherichia coli/drug effects , Humans , Nanoparticles/chemistry , Silver/chemistry , Silver/pharmacology , Staphylococcus aureus/drug effects
5.
Vet Res Commun ; 28(4): 339-46, 2004 May.
Article in English | MEDLINE | ID: mdl-15222738

ABSTRACT

An immunocomb-based dot-ELISA, employing specially designed apparatus, was used to measure the antibody status for the three major poultry diseases--Newcastle disease, infectious bursal disease and infectious bronchitis--in single test sera. Positive samples could be classified into strong, moderate and weak positives by comparison with the colour reaction given by known strong and weak positive serum controls. The simultaneous dot-immunobinding assay gave reproducible results and allowed considerable savings on the cost of reagents compared to liquid ELISA. The antigen-coated immunocomb can be stored under refrigeration and the test can be performed rapidly under field conditions by trained personnel.


Subject(s)
Birnaviridae Infections/veterinary , Chickens , Coronavirus Infections/veterinary , Enzyme-Linked Immunosorbent Assay/veterinary , Newcastle Disease/blood , Poultry Diseases/virology , Animals , Antigens, Viral , Birnaviridae Infections/blood , Birnaviridae Infections/virology , Coronavirus Infections/blood , Coronavirus Infections/virology , Enzyme-Linked Immunosorbent Assay/instrumentation , Enzyme-Linked Immunosorbent Assay/methods , Infectious bronchitis virus/isolation & purification , Infectious bursal disease virus/isolation & purification , Newcastle Disease/virology , Newcastle disease virus/isolation & purification , Poultry Diseases/blood
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