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1.
Indian J Microbiol ; 57(2): 188-194, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28611496

RESUMO

In the present study, gold nanoparticles (AuNPs) synthesis was carried out by using a rare bacteriophage which is morphologically similar to 7-11 phages of the C3 morphotype of tailed phage belonging to Podoviridae family as green route. Effect of various physiological parameters like pH, temperature and concentration of gold chloride salt on AuNPs synthesis was studied. The reaction mixtures have shown vivid colours at various physiological parameters. Phage inspired AuNPs were further characterized by using different techniques such as UV-Vis spectrophotometry, scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), X-ray diffraction (XRD) and dynamic light scattering (DLS). DLS study revealed synthesis of various sizes of AuNPs in the range of 20-100 nm. SEM studies revealed synthesis of varied shaped AuNPs, viz., spheres, hexagons, triangles, rhomboids and rectangular etc. The presence of Au in the nanostructures was confirmed by EDS. The XRD pattern reflects the crystalline nature and nano size of AuNPs. These phage inspired AuNPs showed anti-bacterial activity against different bacterial pathogens. Anti-biofilm activity of AuNPs was evaluated on a glass slide. It was noticed that at 0.2 mM concentration of these AuNPs about 80% of biofilm formation by Pseudomonas aeruginosa, a human pathogen was inhibited. Thus, the phage inspired AuNPs synthesis could be potential therapeutic agents against human pathogens.

2.
Indian J Exp Biol ; 52(11): 1147-51, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25434111

RESUMO

The study was undertaken with the aim of exploring novel and beneficial agro activities of rare actinomycetes like Microbispora sp. V2. The antagonistic activity of Microbispora sp. V2 was evaluated as a biocontrol agents against Sclerotium rolfsii, a soil-borne fungal plant pathogen. The methodology performed for evaluation of biocontrol agent was in vitro evaluation assay which comprised of three tests viz., cellophane overlay technique, seed germination test and Thiram (fungicide) tolerance of Microbispora sp. V2. The isolate was found to inhibit the fungal pathogen Sclerotium rolfsii to 91.43% in cellophane assay. In seed germination assay, Microbispora sp. V2 treated seeds resulted in 25.75% increased germination efficiency, as compared to seeds infected by Sclerotium rolfsii. The isolate Microbispora sp. V2 could tolerate 1000 microg mL(-1) of Thiram (fungicide). The in vitro assay studies proved that Microbispora sp. V2 can be used as antifungal antagonist and thus posses' great potential as biocontrol agent against southern blight caused by Sclerotium rolfsii in Zea mays L (Baby corn) which causes large economical losses.


Assuntos
Actinomycetales/fisiologia , Basidiomycota , Controle Biológico de Vetores/métodos , Doenças das Plantas/prevenção & controle , Zea mays/microbiologia , Actinomycetales/efeitos dos fármacos , Biomassa , Farmacorresistência Bacteriana , Fermentação , Fungicidas Industriais/farmacologia , Germinação , Técnicas In Vitro , Fenazinas/metabolismo , Doenças das Plantas/microbiologia , Sementes/microbiologia , Sementes/fisiologia , Tiram/farmacologia
3.
Can J Microbiol ; 59(5): 318-23, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23647344

RESUMO

A lytic phage of Salmonella serovar Paratyphi B, named φSPB, was isolated from surface waters of the Pavana River in India. Phage φSPB is a member of the Podoviridae family and is morphologically similar to the 7-11 phages of the C3 morphotype of tailed phages, characterized by a very long, cigar-shaped head. The head measured approximately 153 × 57 nm, and the tail size was 12 × 7 nm. The phage was stable over a wide range of pH (4-9) and temperature (4-40 °C). The adsorption rate constant was 4.7 × 10(-10). Latent and eclipse periods were 10 and 15 min, respectively, and the burst size was 100 plaque-forming units/infected cell after 25 min at 37 °C. The phage DNA was 59 kb in size. Ten major proteins were observed on SDS-PAGE, although some of these proteins could be bacterial contaminants. This is the first report of Salmonella enterica subsp. enterica serovar Paratyphi B phage of C3 morphotype from India that has many unique features, such as high replication potential, short replication time, and stability over a wide range of pH and temperature, making it a promising biocontrol agent against the drug-resistant strains of Salmonella Paratyphi B.


Assuntos
Podoviridae/isolamento & purificação , Podoviridae/fisiologia , Rios/virologia , Fagos de Salmonella/isolamento & purificação , Fagos de Salmonella/fisiologia , Salmonella paratyphi B/virologia , Índia , Podoviridae/química , Podoviridae/crescimento & desenvolvimento , Fagos de Salmonella/química , Fagos de Salmonella/crescimento & desenvolvimento , Temperatura
4.
Antonie Van Leeuwenhoek ; 104(1): 25-36, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23588417

RESUMO

The fungal organisms, especially pathogens, change their vegetative (Y, unicellular yeast and H, hypha) morphology reversibly for survival and proliferation in the host environment. NAD-dependent glutamate dehydrogenase (NAD-GDH, EC 1.4.1.2) from a non-pathogenic dimorphic zygomycete Benjaminiella poitrasii was previously reported to be an important biochemical correlate of the transition process. The enzyme was purified to homogeneity and characterized. It is a 371 kDa native molecular weight protein made up of four identical subunits. Kinetic studies showed that unlike other NAD-GDHs, it may act as an anabolic enzyme and has more affinity towards 2-oxoglutarate than L-glutamate. Chemical modifications revealed the involvement of single histidine and lysine residues in the catalytic activity of the enzyme. The phosphorylation and dephosphorylation study showed that the NAD-GDH is present in active phosphorylated form in hyphal cells of B. poitrasii. Two of the 1,2,3 triazole linked ß-lactam-bile acid conjugates synthesized in the laboratory (B18, B20) were found to be potent inhibitors of purified NAD-GDH which also significantly affected Y-H transition in B. poitrasii. Furthermore, the compound B20 inhibited germ tube formation during Y-H transition in Candida albicans strains and Yarrowia lipolytica. The possible use of NAD-GDH as a target for antifungal agents is discussed.


Assuntos
Proteínas Fúngicas/isolamento & purificação , Glutamato Desidrogenase/isolamento & purificação , Mucorales/enzimologia , Cloreto de Amônio/metabolismo , Antifúngicos/síntese química , Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Candida albicans/enzimologia , Candida albicans/ultraestrutura , Catálise , Cromatografia em Agarose , Avaliação Pré-Clínica de Medicamentos , Proteínas Fúngicas/antagonistas & inibidores , Proteínas Fúngicas/metabolismo , Glutamato Desidrogenase/antagonistas & inibidores , Glutamato Desidrogenase/metabolismo , Ácido Glutâmico/metabolismo , Histidina/química , Histidina/efeitos dos fármacos , Hifas/enzimologia , Ponto Isoelétrico , Ácidos Cetoglutáricos/metabolismo , Lisina/química , Lisina/efeitos dos fármacos , Terapia de Alvo Molecular , Peso Molecular , Mucorales/efeitos dos fármacos , Mucorales/fisiologia , Mucorales/ultraestrutura , NAD/metabolismo , Fosforilação , Processamento de Proteína Pós-Traducional , Especificidade por Substrato , Triazóis/farmacologia , Yarrowia/efeitos dos fármacos , Yarrowia/enzimologia , Yarrowia/ultraestrutura
5.
PLoS One ; 6(3): e17491, 2011 Mar 02.
Artigo em Inglês | MEDLINE | ID: mdl-21399689

RESUMO

Botulinum neurotoxin (BoNT) potently inhibits cholinergic signaling at the neuromuscular junction. The ideal countermeasures for BoNT exposure are monoclonal antibodies or BoNT antisera, which form BoNT-containing immune complexes that are rapidly cleared from the general circulation. Clearance of opsonized toxins may involve complement receptor-mediated immunoadherence to red blood cells (RBC) in primates or to platelets in rodents. Methods of enhancing immunoadherence of BoNT-specific antibodies may increase their potency in vivo. We designed a novel fusion protein (FP) to link biotinylated molecules to glycophorin A (GPA) on the RBC surface. The FP consists of an scFv specific for murine GPA fused to streptavidin. FP:mAb:BoNT complexes bound specifically to the RBC surface in vitro. In a mouse model of BoNT neutralization, the FP increased the potency of single and double antibody combinations in BoNT neutralization. A combination of two antibodies with the FP gave complete neutralization of 5,000 LD50 BoNT in mice. Neutralization in vivo was dependent on biotinylation of both antibodies and correlated with a reduction of plasma BoNT levels. In a post-exposure model of intoxication, FP:mAb complexes gave complete protection from a lethal BoNT/A1 dose when administered within 2 hours of toxin exposure. In a pre-exposure prophylaxis model, mice were fully protected for 72 hours following administration of the FP:mAb complex. These results demonstrate that RBC-targeted immunoadherence through the FP is a potent enhancer of BoNT neutralization by antibodies in vivo.


Assuntos
Anticorpos Monoclonais/imunologia , Anticorpos Neutralizantes/imunologia , Toxinas Botulínicas Tipo A/imunologia , Eritrócitos/metabolismo , Testes de Neutralização/métodos , Proteínas Recombinantes de Fusão/metabolismo , Animais , Anticorpos Monoclonais/metabolismo , Biotinilação , Toxinas Botulínicas Tipo A/sangue , Feminino , Injeções , Camundongos , Ligação Proteica
6.
Bioresour Technol ; 100(21): 5132-9, 2009 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-19539465

RESUMO

Feasibility of using chocolate industry wastewater as a substrate for electricity generation using activated sludge as a source of microorganisms was investigated in two-chambered microbial fuel cell. The maximum current generated with membrane and salt bridge MFCs was 3.02 and 2.3 A/m(2), respectively, at 100 ohms external resistance, whereas the maximum current generated in glucose powered MFC was 3.1 A/m(2). The use of chocolate industry wastewater in cathode chamber was promising with 4.1 mA current output. Significant reduction in COD, BOD, total solids and total dissolved solids of wastewater by 75%, 65%, 68%, 50%, respectively, indicated effective wastewater treatment in batch experiments. The 16S rDNA analysis of anode biofilm and suspended cells revealed predominance of beta-Proteobacteria clones with 50.6% followed by unclassified bacteria (9.9%), alpha-Proteobacteria (9.1%), other Proteobacteria (9%), Planctomycetes (5.8%), Firmicutes (4.9%), Nitrospora (3.3%), Spirochaetes (3.3%), Bacteroides (2.4%) and gamma-Proteobacteria (0.8%). Diverse bacterial groups represented as members of the anode chamber community.


Assuntos
Bactérias/citologia , Fontes de Energia Bioelétrica/microbiologia , Cacau/química , Resíduos Industriais , Esgotos/microbiologia , Eliminação de Resíduos Líquidos , Purificação da Água , Bactérias/metabolismo , Células Clonais , Conservação de Recursos Energéticos , Meios de Cultura , Eletricidade , Eletrodos/microbiologia , Eletrólitos , Glucose/metabolismo , Membranas Artificiais , Filogenia , Proteobactérias/citologia , Proteobactérias/genética , Prótons , Cloreto de Sódio/química
7.
PLoS One ; 3(8): e3023, 2008 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-18714390

RESUMO

BACKGROUND: Botulinum neurotoxins (BoNT) are a family of category A select bioterror agents and the most potent biological toxins known. Cloned antibody therapeutics hold considerable promise as BoNT therapeutics, but the therapeutic utility of antibodies that bind the BoNT light chain domain (LC), a metalloprotease that functions in the cytosol of cholinergic neurons, has not been thoroughly explored. METHODS AND FINDINGS: We used an optimized hybridoma method to clone a fully human antibody specific for the LC of serotype A BoNT (BoNT/A). The 4LCA antibody demonstrated potent in vivo neutralization when administered alone and collaborated with an antibody specific for the HC. In Neuro-2a neuroblastoma cells, the 4LCA antibody prevented the cleavage of the BoNT/A proteolytic target, SNAP-25. Unlike an antibody specific for the HC, the 4LCA antibody did not block entry of BoNT/A into cultured cells. Instead, it was taken up into synaptic vesicles along with BoNT/A. The 4LCA antibody also directly inhibited BoNT/A catalytic activity in vitro. CONCLUSIONS: An antibody specific for the BoNT/A LC can potently inhibit BoNT/A in vivo and in vitro, using mechanisms not previously associated with BoNT-neutralizing antibodies. Antibodies specific for BoNT LC may be valuable components of an antibody antidote for BoNT exposure.


Assuntos
Anticorpos Monoclonais , Antitoxina Botulínica/imunologia , Toxinas Botulínicas/antagonistas & inibidores , Toxinas Botulínicas/imunologia , Cadeias Leves de Imunoglobulina , Especificidade de Anticorpos , Linhagem Celular Tumoral , Clonagem Molecular , Humanos , Cadeias Leves de Imunoglobulina/genética , Cinética , Neuroblastoma , Proteínas Recombinantes/imunologia , Sorotipagem
8.
J Appl Microbiol ; 103(6): 2622-8, 2007 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-17850302

RESUMO

AIM: The aim of this study was to isolate and identify antifungal lactic acid bacteria from fresh vegetables, and evaluate their potential in preventing fungal spoilage of vegetables. METHODS AND RESULTS: Lactic acid bacteria from fresh vegetables were enriched in MRS (de Man Rogosa Sharpe) broth and isolated by plating on MRS agar. All the isolates (359) were screened for activity against Aspergillus flavus of which 10% showed antifungal activity. Potent antifungal isolates were identified by phenotypic characters and confirmed by partial 16S rRNA gene sequencing. These were screened against additional spoilage fungi viz. Fusarium graminearum, Rhizopus stolonifer, Sclerotium oryzae, Rhizoctonia solani, Botrytis cinerea and Sclerotinia minor by overlay method. Most of the isolates inhibited wide range of spoilage fungi. When fresh vegetables were inoculated with either cell suspension (10(4) cells ml(-1)) or cell-free supernatant of Lact. plantarum, followed by application of vegetable spoilage fungi (A. flavus and F. graminearum, R. stolonifer, B. cinerea each with 10(4) conidia ml(-1)) the vegetable spoilage was significantly delayed than control. CONCLUSIONS: Fresh vegetables constitute a good source of lactic acid bacteria with ability to inhibit wide range of spoilage fungi. Such bacteria can be applied to enhance shelf-life of vegetables. In the present study, we report for the first time the antifungal activity of Weissella paramessenteroides and Lact. paracollinoides isolated from fresh vegetables, against wide range of food spoilage fungi. SIGNIFICANCE AND IMPACT OF THE STUDY: Fresh vegetables can be used as a source of antifungal lactic acid bacteria. Their exploitation as biopreservative will help in prolonging shelf-life of fresh vegetables.


Assuntos
Microbiologia de Alimentos , Conservação de Alimentos , Fungos/fisiologia , Lactobacillaceae/fisiologia , Verduras , Antibiose , Aspergillus fumigatus , Botrytis , Fusarium , Genes Bacterianos , Lactobacillaceae/genética , Lactobacillaceae/isolamento & purificação , Testes de Sensibilidade Microbiana , Rhizoctonia , Ribotipagem
9.
Lett Appl Microbiol ; 44(6): 588-94, 2007 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-17576218

RESUMO

AIMS: To study the occurrence and diversity of Salmonella serovars in urban water supply systems of Nepal. METHODS AND RESULTS: Occurrence of Salmonella was detected in 42 out of 300 water samples by enrichment culture technique in selenite F broth followed by plating on Salmonella Shigella agar. A total of 54 isolates identified to genus level by standard tests were subsequently confirmed by serotyping, phage typing and PCR detection of virulence genes (inv A and spv C). The predominant serotype was Salmonella Typhimurium, followed by Salm. Typhi, Salm. Paratyphi A and Salmonella Enteritidis. Most of the Salm. Typhi isolates were E1 phage type followed by UVS4, A and UVS1. All isolates of Salm. Paratyphi A and Salm. Enteritidis were an untypable (UT) phage type. The majority of isolates were multi-drug resistant as revealed by Kirby-Bauer disc diffusion technique. Ceftriaxone resistant isolates of Salm. Enteritidis indicated the presence of one of the ESBL genes, blaSHV, whereas the genes blaTEM and blaCTX were absent. CONCLUSIONS: The microbiological quality of the urban water supply is poor and indicates possibility of fatal outbreaks of enteric fever and related infections in Nepal. SIGNIFICANCE AND IMPACT OF THE STUDY: The present study will be useful in water borne disease control and prevention strategy formulation in Nepal and in the global context.


Assuntos
Salmonella/classificação , Salmonella/isolamento & purificação , Microbiologia da Água , Abastecimento de Água , Antibacterianos/farmacologia , Tipagem de Bacteriófagos , Surtos de Doenças/prevenção & controle , Farmacorresistência Bacteriana , Testes de Sensibilidade Microbiana , Nepal , Reação em Cadeia da Polimerase , Salmonella/efeitos dos fármacos , Infecções por Salmonella/prevenção & controle , Sorotipagem
10.
Pak J Biol Sci ; 10(9): 1519-24, 2007 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-19069968

RESUMO

A total of 455 domestic animals (cow, horse and camel) and poultry from south of Iran were surveyed for fecal carriage of Campylobacter spp. Out of all collected fecal samples, the highest isolation rate of Campylobacter was recorded among poultry (35%), followed by horse (27%) and cow (21%) while, lowest isolation rate was recorded among camel. Of the 85 Campylobacter strains isolated, 76 were classified as catalase positive Campylobacter. Out of them, high frequency of occurrence was belonged to Campy. jejuni. Furthermore, catalase positive Campylobacter spp. were isolated from all the sources of investigation, other than camel. The results obtained from biotyping of the isolates indicated Camp. lari biotype I followed by Camp. jejuni and Camp. coli biotypes I existed in high frequency; while Camp. jejuni biotype II and untypable Campylobacter existed in low frequency. Overall, domestic animals and poultry other than camels are vehicle of Campylobacter in the area of investigation therefore, the people who living in this area may be infected via feces of domestic animals and poultry.


Assuntos
Camelus/microbiologia , Campylobacter/isolamento & purificação , Bovinos/microbiologia , Cavalos/microbiologia , Aves Domésticas/microbiologia , Animais , Técnicas de Tipagem Bacteriana , Campylobacter/classificação , Infecções por Campylobacter/epidemiologia , Infecções por Campylobacter/veterinária , Dieta , Fezes/microbiologia , Humanos , Irã (Geográfico)/epidemiologia
11.
Lett Appl Microbiol ; 43(1): 84-90, 2006 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-16834726

RESUMO

AIM: To determine the effect of two physiologically important temperatures on growth and chemotaxis in Campylobacter jejuni. METHODS AND RESULTS: Growth curves of Camp. jejuni were compared at 37 degrees C and 42 degrees C. Chemotaxis was compared at 37 degrees C and 42 degrees C by the disc and capillary assays. Student's t-test was applied to the results of the capillary assay to assess the significance in the difference between chemotaxis at the two temperatures. Both, the growth rate and chemotactic ability of the isolate, were found to be greater at 37 degrees C. CONCLUSIONS: Quorum sensing (related to population density), a regulation mechanism of virulence in micro-organisms, has been reported in Campylobacter. Chemotaxis is also a known virulence factor of Campylobacter. Both, growth (in terms of population density) and chemotaxis, being greater at 37 degrees C than at 42 degrees C, suggests that the physiological temperature of humans (37 degrees C) might be more favourable for the expression of virulence in Campylobacter than that of birds (42 degrees C). SIGNIFICANCE AND IMPACT OF THE STUDY: It is as yet not known why Campylobacter causes disease in humans but is avirulent in birds. This study suggests that the human body temperature is optimum for growth and chemotaxis in Campylobacter. There is scope for the study of temperature regulation of other virulence determinants of Campylobacter.


Assuntos
Campylobacter jejuni/crescimento & desenvolvimento , Campylobacter jejuni/fisiologia , Quimiotaxia , Aminoácidos/farmacologia , Animais , Técnicas Bacteriológicas , Ácidos e Sais Biliares/farmacologia , Campylobacter jejuni/efeitos dos fármacos , Campylobacter jejuni/patogenicidade , Capilares , Carboidratos/farmacologia , Quimiotaxia/efeitos dos fármacos , Meios de Cultura/química , Humanos , Temperatura , Virulência
12.
Bioresour Technol ; 97(12): 1377-81, 2006 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-16115759

RESUMO

Bioremediation potential of Phanerochaete chrysosporium strains NCIM 1073, NCIM 1106 and NCIM 1197 to decolourise molasses in solid and liquid molasses media was studied. Strains varied in the pattern of molasses decolourisation on solid medium by Giant colony method. Under submerged cultivation conditions, strain NCIM 1073 did not decolourise molasses while, strains NCIM 1106 and NCIM 1197 could decolourise molasses up to 82% and 76%, respectively. Under stationary cultivation conditions, none of the strains could decolourise molasses. This was overcome by increasing the surface area of the culture in flat bottom glass bottles under stationary cultivation conditions. Under submerged cultivation conditions, growth was more or less same in all strains. However, the lignin peroxidase and manganese peroxidase activities were significantly less in the strain NCIM 1073. Under stationary cultivation conditions, none of the strains could produce enzymes lignin peroxidase, manganese peroxidase and laccase. However, all of them could produce lignin peroxidase and manganese peroxidase when cultivated in flat bottom glass bottles under stationary cultivation conditions.


Assuntos
Biotecnologia/métodos , Melaço , Phanerochaete/metabolismo , Biodegradação Ambiental , Catálise , Concentração de Íons de Hidrogênio , Lacase/química , Modelos Biológicos , Peroxidases/química , Fatores de Tempo
13.
Indian J Exp Biol ; 43(8): 751-4, 2005 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-16121720

RESUMO

To study antimicrobial activity of shallot in comparison with that of garlic and onion against 23 strains of fungi and bacteria, water extracts of garlic, shallot and onion bulbs were prepared. Each extract was studied in different forms for their antimicrobial activity viz., fresh extract, dry extract and autoclaved extract. Minimal inhibitory concentration and minimal lethal concentrations of these extracts were determined against all organisms by broth dilution susceptibility test. Fresh extract of garlic showed greater antimicrobial activity as compared to similar extracts of onion and shallot. However, dried and autoclaved extracts of shallot showed more activity than similar extracts of onion and garlic. Fungi were more sensitive to shallot extract than bacteria. Amongst bacteria, B. cereus was most sensitive (MIC=5 mg ml(-1)). The lowest minimum bactericidal concentration of shallot extract amongst bacteria tested was 5 mg ml(-1) for B. cereus. Amongst fungi, Aureobasidium pullulans and Microsporum gypseum were most sensitive (MIC= 0.15 mg ml(-1)). The lowest minimum lethal concentration was 2.5 mg ml(-1) for Microsporum gypseum and Trichophyton mentagrophytes. It was therefore, expected that the antimicrobial principle of shallot was different than the antimicrobial compounds of onion and garlic. In addition, the antimicrobial component of the shallot extract was stable at 121 degrees C.


Assuntos
Antibacterianos/farmacologia , Antifúngicos/farmacologia , Cebolinha Branca/química , Alho/química , Testes de Sensibilidade Microbiana , Cebolas/química , Extratos Vegetais/farmacologia
14.
Indian J Med Microbiol ; 23(1): 48-51, 2005 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-15928423

RESUMO

Environmental samples were subjected to determine frequency of occurrence of pathogenic campylobacters in the environment. The antimicrobial susceptibility of the isolates was tested to evaluate the level of antibiotic sensitive campylobacters in the environment of investigation. In all, 70 Campylobacter isolates were obtained from water and domestic animal faeces samples using Kapadnis-Baseri device and antimicrobial susceptibility of them was determined by disc diffusion test and E- test. The results indicated that all the isolates of Campylobacter were sensitive to ciprofloxacin and resistant to cefotaxime, cephalexin and ampicillin. Lowest MIC values were observed for ciprofloxacin and gentamicin (2 microg/mL) and highest MIC values for ampicillin and chloramphinicol (256 microg/mL). In general, pathogenic Campylobacter spp. were prevalent in large numbers in the environment, however, they were sensitive to ciprofloxacin.


Assuntos
Antibacterianos/farmacologia , Campylobacter/efeitos dos fármacos , Campylobacter/genética , Campylobacter/isolamento & purificação , Farmacorresistência Bacteriana , Testes de Sensibilidade Microbiana/métodos
15.
J Biosci ; 30(2): 201-11, 2005 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-15886456

RESUMO

In all 312 actinomycete strains were isolated from water and soil samples from different regions. All these isolates were purified and screened for their antifungal activity against pathogenic fungi. Out of these, 22% of the isolates exhibited activity against fungi. One promising strain, Streptomyces albidoflavus PU 23 with strong antifungal activity against pathogenic fungi was selected for further studies. Antibiotic was extracted and purified from the isolate. Aspergillus spp. was most sensitive to the antibiotic followed by other molds and yeasts. The antibiotic was stable at different temperatures and pH tested and there was no significant loss of the antifungal activity after treatment with various detergents and enzymes. Synergistic effect was observed when the antibiotic was used in combination with hamycin. The antibiotic was fairly stable for a period of 12 months at 4 degree C. The mode of action of the antibiotic seems to be by binding to the ergosterol present in the fungal cell membrane resulting in the leakage of intracellular material and eventually death of the cell. The structure of the antibiotic was determined by elemental analysis and by ultraviolet (UV), Fourier transform infrared (FTIR), nuclear magnetic resonance (NMR) and liquid chromatography mass spectra (LCMS). The antibiotic was found to be a straight chain polyhydroxy, polyether, non-proteinic compound with a single double bond, indicating a nonpolyene antifungal antibiotic.


Assuntos
Antifúngicos/isolamento & purificação , Microbiologia do Solo , Streptomyces/química , Microbiologia da Água , Antifúngicos/metabolismo , Antifúngicos/toxicidade , Cromatografia Líquida , Ergosterol/metabolismo , Fungos/efeitos dos fármacos , Concentração de Íons de Hidrogênio , Índia , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Polienos/toxicidade , Espectroscopia de Infravermelho com Transformada de Fourier , Temperatura , Raios Ultravioleta
16.
Indian J Med Res ; 121(3): 164-70, 2005 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-15802758

RESUMO

BACKGROUND & OBJECTIVE: Dermatophytes responsible for causing dermatophytoses in humans have acquired resistance to certain antimycotic drugs. We isolated naturally occurring actinomycetes with an ability to produce metabolites having antimycotic property. The timecourse of antifungal metabolite production in terms of arbitrary units (AU) under optimum conditions was studied. METHODS: Water and soil samples were collected from various locations. The actinomycetes were isolated on starch casein medium and screened for their antifungal activity against yeasts and molds including dermatophytes. One promising isolate which showed a unique, stable and interesting property of inhibiting only dermatophytes was selected and characterized. Optimization of antifungal metabolite production in terms of AU using Trichphyton rubrum as target was done. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values of the culture supernatant from the isolate and that of griseofulvin were determined for all dermatophytes. RESULTS: Of the 218 actinomycete isolates, 14 per cent produced the metabolites having antifungal activity. The selected actinomycete, identified as Streptomyces rochei AK 39 produced metabolite, which was active against only dermatophytes whereas yeasts and other molds were resistant to it. Starch casein medium was found to be good for inducing antifungal activity in the isolate. The maximum antifungal metabolite production (400 AU/ml) was achieved in the late log phase, which remained constant during the stationery phase, and it was extracellular in nature. The MIC and MFC values of the culture supernatant from the isolate against the dermatophytes were within the range 1.25 to 5 and 1.25 to 10 AU/ml respectively. INTERPRETATION & CONCLUSION: The metabolite from Streptomyces rochei AK 39 was produced during late log phase and was active against only dermatophytes with a greater potency than griseofulvin. However, this needs further investigation using purified powdered form of the active component.


Assuntos
Antibacterianos/farmacologia , Arthrodermataceae/efeitos dos fármacos , Streptomyces/metabolismo , Actinobacteria/crescimento & desenvolvimento , Actinobacteria/isolamento & purificação , Actinobacteria/metabolismo , Antibacterianos/metabolismo , Arthrodermataceae/patogenicidade , Dermatomicoses/tratamento farmacológico , Farmacorresistência Fúngica , Griseofulvina/farmacologia , Humanos , Técnicas In Vitro , Testes de Sensibilidade Microbiana , Microbiologia do Solo , Streptomyces/crescimento & desenvolvimento , Streptomyces/isolamento & purificação , Microbiologia da Água
17.
Indian J Exp Biol ; 42(9): 928-32, 2004 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-15462189

RESUMO

About 312 actinomycetes were isolated from soil samples on chitin agar. All these isolates were purified and screened for their antifungal activity against pathogenic fungi. Out of these, 22% of the isolates exhibited activity against fungi. One promising isolate with strong antifungal activity against pathogenic fungi was selected for further studies. This isolate was from Pune, and was active against both yeasts and molds. Various fermentation parameters were optimized. Based on morphological and biochemical parameters, the isolate was identified as Streptomyces. The correlation of antifungal activity with growth indicated growth dependent production of antimetabolite. Maximum antifungal metabolite production (600 units/ml) was achieved in the late log phase, which remained constant during stationery phase, and it was extracellular in nature.


Assuntos
Actinobacteria/isolamento & purificação , Antibiose , Antifúngicos/farmacologia , Fungos/efeitos dos fármacos , Fungos/crescimento & desenvolvimento , Microbiologia do Solo , Actinobacteria/crescimento & desenvolvimento , Testes de Sensibilidade Microbiana/métodos , Solo/análise , Streptomyces/crescimento & desenvolvimento , Streptomyces/isolamento & purificação
18.
J Appl Microbiol ; 97(4): 853-60, 2004.
Artigo em Inglês | MEDLINE | ID: mdl-15357735

RESUMO

AIMS: To develop a method that involves sample processing, and blood- and antibiotic-free medium for isolation and enumeration of Campylobacter spp. from environmental samples. METHODS AND RESULTS: The sample processing (preT) was standardized to minimize the population of competing bacteria. A blood- and antibiotic-free differential, Kapadnis-Baseri medium (KB medium) was formulated and tested for isolation of Campylobacter spp. in comparison with CAT medium. PreT-KB method was evaluated in comparison with the conventional viable count method and with the conventional most probable number (C. MPN) method for enumeration of Campylobcater from environmental samples. The results indicated that sample processing significantly reduced population of competing bacteria. The KB medium selected Gram-negative bacteria and differentiated Campylobacter from lactose-fermenting competing bacteria. The population of Campylobacter detected by preT-KB method was similar to that by conventional viable count method. While, the population of Campylobacter spp. determined by preT-KB method was higher than that by C. MPN method. In addition, the preT-KB method detected antibiotic sensitive campylobacters. CONCLUSION: The preT minimizes population of competing bacteria and the KB medium selects Gram-negative bacteria and differentiates Campylobacter from them. Therefore, Campylobacter can be isolated from environmental samples without using antibiotics. SIGNIFICANCE AND IMPACT OF THE STUDY: The preT-KB method is simple and facilitates isolation of antibiotic sensitive and enumeration of Campylobacter in the environmental samples. Therefore, the new method will be useful for isolation and enumeration of Campylobacter from water, food and sewage samples. Besides, it would also detect antibiotic-sensitive campylobacters, which are not detected by conventional viable count and MPN methods.


Assuntos
Técnicas Bacteriológicas/métodos , Campylobacter/isolamento & purificação , Animais , Antibacterianos/farmacologia , Búfalos , Campylobacter/efeitos dos fármacos , Bovinos , Contagem de Colônia Microbiana/métodos , Meios de Cultura , Resistência Microbiana a Medicamentos , Fezes/microbiologia , Esgotos/microbiologia , Ovinos
19.
J Appl Microbiol ; 96(4): 664-70, 2004.
Artigo em Inglês | MEDLINE | ID: mdl-15012803

RESUMO

AIM: To design a special device which can be used with nonselective blood-free nutrient agar without enrichment for detection of campylobacters from water. METHODS AND RESULTS: The Kapadnis-Baseri device (KB device) was designed and evaluated in comparison with the conventional method (C method) for detection of Campylobacter spp. from river water samples. The results indicated that the recovery of Campylobacter spp. by KB device was relatively more than by C method. CONCLUSIONS: To date, the methods for recovery of campylobacters are time consuming and involve use of special culture media, which is cost ineffective. The KB device is designed based on two important characters of Campylobacter, viz. motility and activity at low temperature. With this device we isolated Campylobacter spp. from river water on nonselective media without enrichment. Thus this device is as effective as the use of antibiotic media for the isolation of campylobacters. SIGNIFICANCE AND IMPACT OF THE STUDY: The KB device will be useful for isolation of Campylobacter spp. from water and other environmental samples, which is less time consuming and inexpensive. Besides, this device allows isolation of antibiotic sensitive campylobacters.


Assuntos
Campylobacter/isolamento & purificação , Microbiologia da Água , Poluição da Água , Técnicas Bacteriológicas , Campylobacter/fisiologia , Movimento , Rios
20.
Indian J Med Microbiol ; 22(1): 39-43, 2004.
Artigo em Inglês | MEDLINE | ID: mdl-17642684

RESUMO

PURPOSE: Campylobacter spp. is a major food borne pathogen and shows resistance towards gamma radiation. In the present study, effect of gamma radiation was assessed on the indigenous strains of Campylobacter spp. inoculated in food and water samples. METHODS: Campylobacter spp. were isolated from river water and faeces of various birds and animals. The growth rate was studied for these isolates by propagating them in Kapadnis-Baseri medium. The survival of Campylobacter spp. inoculated in food and water samples was tested after exposing them to gamma radiation. RESULTS: The isolates survived well in meat and milk samples and were sensitive to 1.8 KGy dose of gamma radiation, which lies with in the FDA limit. The effect of radiation on Campylobacter spp. varied with the species and the type of food. CONCLUSIONS: The results obtained suggest that the dose of gamma radiation should be standardized depending on the Campylobacter spp. and the type of food that is being processed.

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