full issue - Association of Biotechnology and Pharmacy

More documents

Recommendations

Info

Current Trends in Biotechnology and PharmacyVol. 5 (1) 1043-1053 January 2011. ISSN 0973-8916 (Print), 2230-7303 (Online)1045Serratia liquefaciens (JNMC), Pseudomonasaureginosa (JNMC), Penicillum crysogenum(JNMC), Candida albicans (JNMC-1),Candida albicans (JNMC-2) and from ITRC(Indian Toxicology Research Center, Lucknow)Bacillus cereus (ATCC 10876), Escherichia coli(ATCC 35218). Bacterial cultures were grownat 37°C on nutrient agar medium and fungi weregrown at 28°C on Sabouraud’s dextrose agar. Allthe cultures were preserved at 4°C and subculturedregularly.Morphological, Physiological and biochemicalcharacteristics: The potent Streptomycesisolates selected after primary screening wascharacterized by morphological, biochemical andphysiological studies (Table III). They were thenobserved for their mycelia structure, and sporearrangements on the mycelia under lightmicroscope at 1000X resolution by using Gramstaining. The observed morphology of the isolateswas compared with the actinomycetesmorphology provided in Bergey’s Manual for thepresumptive identification of the isolates. Thebiochemical tests performed were catalase,lecithinase (egg yolk rection), oxidase, citrateutilization, nitrate reduction, starch hydrolysis,tween-20 hydrolysis (lipolysis), urea hydrolysis,gelatin hydrolysis, ability to produce H 2S,degradation of elastin, arbutin, pectin, guanine,tyrosine, esculin and xanthine. Carbon utilizationsuch as D-glucose, D-fructose, D-galactose, D-xylose, D-mannitol, D-sorbitol, D-lactose, D-arabinose, D-cellubiose, meso-inositol, dextran,melibiose, melezitose, xylitol, L-rhamnose, L-raffinose was determined on plates containingbasal salt agar media (KH 2PO 42.64 g,K 2HPO 4.3H 2O 5.65 g, (NH4) 2SO 42.64 g, tracesalt solution (CuSO 4.5H 2O 0.64 g, FeSO 4.7H 2O0.11 g, MnCl 2.4H 2O 0.7 g, ZnSO 4.7H 2O 0.15 g,distilled water 100ml) 1ml, agar 20 g and distilledwater 1000 ml) to which carbon sources wereadded to a final concentration of 1.0% (14). Theplates were incubated at 28°C and growth wasassessed after 7, 14, 21 days using glucose aspositive control and carbon source free mediumas negative control. The ability to utilize nitrogensources such as L-cystein, L-valine, L-leucine,L-asparagine, L-hydroxyproline, L-serine, L-phenylalanine and L-histidine was determined ina basal medium (glucose 1.0 g, MgSO 4.7H 2O0.05 g, FeSO 4.7H 2O 0.001 g, K 2HPO 40.01 g,NaCl 0.05 g, agar 2.0 g and distilled water 100ml) to which 0.1% nitrogen sources were added.The plates were incubated at 28°C and growthwas recorded after 7, 14, 21 days using L-asparagine as positive control and a mediumwithout nitrogen source as a negative control (16).Determination of cell wall L, L-diaminopimelicacid was performed by the method of Becker etal., (15). Antibiotic sensitivity against the differentantibiotics (Himedia, Mumbai, India) gentamycin(G 30 ), chloromphenicol (C 10 ), linezolid (LZ 30 ),ciprofloxacin (CF 5 ), amoxicillin (AM 10 ),methicillin(M 10 ), nalidixic acid (NA 30 ), novobiocin(Nv 5 ), tetracycline (T 10 ), oleondamycin (OL 15 ),streptomycin (S 25 ), vancomycin (VA 5 ),kanamycin (K 30 ), polymixim B (PB 100 ), neomycin(N 30 ), rifamycin (R 5 ), erythromycin (E 15 ),cephalosporin (CR 30 ), penicillin G (P 10 ),amphotericin B (AP 30 ), fluconazole (F 10 ),miconazole (MIC 30 ) were performed using MullerHinton agar medium by paper disc method (16).ResultsThe genus Streptomyces, which is the mostabundant and a recoverable actinomycete groupisolated from the soil. Total fifty sevenStreptomyces isolates were recorded from thesoil of Durg District of Chhattisgarh according todifferences in colony morphology (17) on starchcasein nitrate agar media (Fig. 1). Out of 57isolates, only 19 isolates showed the antimicrobialactivity. Among this 19 isolates, 11 showed bothIsolation and Characterization of Streptomyces sp.
Current Trends in Biotechnology and PharmacyVol. 5 (1) 1043-1053 January 2011. ISSN 0973-8916 (Print), 2230-7303 (Online)1046Fig. 1. Growth pattern of bioactive Streptomyces isolates on starch casein nitrate agar media fromvarious location of Durg.antibacterial and antifungal activity while 8exhibited only antibacterial activity. The diameterof zone of inhibition (in millimeter) showed by theculture filtrate of 19 isolates is represented in table(1). In present study results indicated that graycolored isolates were showing activity against themost of the test organisms when subjected in tosubmerged culture. The yellow colored isolates(Y8, MS38) and grey colored isolates (A8, MS02and D15) exhibited highest antibacterial andantifungal activity (Fig. 3). Among these isolateY8 (yellow) recorded to show broad spectrumactivity against all gram positive bacteria, gramnegative bacteria and fungi. Results of antibioticsensitivity test was presented in table-2 showedthat Bacillus cereus (MTCC 1305) was the mostsensitive organism inhibited by the extract of 14isolates followed by Bacillus subtilis (MTCC1789), Salmonella typhi (MTCC 531),Escherichia coli (MTCC 1667), Bacillus cereus(ATCC 10876), Staphylococcus aureus (MTCC96), Proteus vulgaris (MTCC 1771), Candidaalbicans (MTCC 1637), Klebsiella Pneumoniae(MTCC 2405), Penicillium citrinum (MTCC1751), Staphylococcus epidermis (MTCC 435),Alternaria alternate (MTCC 1779),Escherichia coli (ATCC 35218), Aspergillusniger (MTCC 872), Staphylococcus aureus(JNMC), Serratia liquefaciens (JNMC)Penicillium crysogenum (JNMC), Candidaalbicans (JNMC-1), Candida albicans (JNMC-2), Bacillus megaterium (MTCC 1684),Pseudomonas aureginosa (JNMC). Antibioticsensitivity profile of isolates is shown table-(II)Most of the isolates showed resistance to theamoxicillin (AM 10 ), methicillin (M 10 ), nalidixicacid (NA 30 ), cephalosporin (CR 30 ), penicillinG(P 10 ), amphotericin B (AP 30 ), fluconazole (F 10 )and showed sensitivity towards the gentamycin(G 30 ), linezolid (LZ 30 ), streptomycin (S 25 ),Singh and Rai
Page 3 and 4:
ISSN 0973-8916Current Trends inBiot
Page 5 and 6:
Current Trends in Biotechnology and
Page 7 and 8:
Page 9 and 10:
Page 11 and 12:
Page 13 and 14:
Page 15 and 16:
Page 17 and 18:
Page 19 and 20:
Page 21 and 22:
Page 23 and 24:
Page 25 and 26:
Page 27 and 28:
Page 29 and 30: Current Trends in Biotechnology and
Page 79: Current Trends in Biotechnology and
Page 123: Current Trends in Biotechnology and
show all

full issue - Association of Biotechnology and Pharmacy

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?