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Isolation, Identification and Evaluation of Antimicrobial Activity of Streptomyces flavogriseus, strain ACTK2 from Soil Sample of Kodagu, Karnataka State (India).

ABSTRACT: BACKGROUND:The search for novel antibiotics continues to be of immense importance in research programs around the world for pharmaceutical, industrial and agricultural applications. Filamentous soil bacteria, belonging to the Streptomyces genus, are widely used as an important biological tool for their ability to produce a wide range of novel secondary metabolites, such as "antibiotics". OBJECTIVES:The aim of the present study was to isolate and identify a strain of Streptomyces with high antibiotic production capability. MATERIALS AND METHODS:The soil sample was collected randomly from the agricultural land of Kushalnagar Taluk of Kodagu district, Karnataka, India. The ACTK2 strain was isolated by serial dilution method and identified based on cultural, morphological, microscopic, biochemical and sequence analysis of 16S rRNA gene parameters. The isolated ACTK2 was analyzed for antimicrobial activities by perpendicular streak and disc diffusion methods, against the Gram-positive bacteria Staphylococcus aureus (MTCC 96), Bacillus subtilis (MTCC 121), Gram-negative Escherichia. coli (MTCC 729), Enterococcus aerogenes (MTCC 2829) and filamentous fungi (Trichoderma harizianum (MTCC6046), Fusarium proliferatum (MTCC 9375). Further, an antimicrobial metabolite from the ACTK2 strain was extracted by solvent extraction method, using n-butanol. The production of the antimicrobial compound by the ACTK2 strain was optimized by using different nutritional media and cultural conditions. RESULTS:The strain Streptomyces flavogriseus designated as ACTK2 (Accession number KC990785) isolated from the soil sample of Kushalnagar Taluk, Kodagu, Karnataka, India, exhibited a broad spectrum of antimicrobial activity against test microorganisms. The optimum growth and antimicrobial compound production by strain ACTK2 was found to be a maximal pH 8, in the shaker incubator at 28ºC, for a period of 10 days. CONCLUSIONS:The crude n-butanol extract of the ACTK2 strain of S. flavogriseus showed a broad spectrum of antimicrobial activities against the test organisms and this opened further research investigations on purification and structural characterization of the active compounds from the crude extract.

SUBMITTER: Khandan Dezfully N

PROVIDER: S-EPMC4376976 | biostudies-literature | 2015 Feb

REPOSITORIES: biostudies-literature

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Isolation, Identification and Evaluation of Antimicrobial Activity of Streptomyces flavogriseus, strain ACTK2 from Soil Sample of Kodagu, Karnataka State (India).

Khandan Dezfully Nooshin N Gottravalli Ramanayaka Janardhana J

Jundishapur journal of microbiology 20150210 2

<h4>Background</h4>The search for novel antibiotics continues to be of immense importance in research programs around the world for pharmaceutical, industrial and agricultural applications. Filamentous soil bacteria, belonging to the Streptomyces genus, are widely used as an important biological tool for their ability to produce a wide range of novel secondary metabolites, such as "antibiotics".<h4>Objectives</h4>The aim of the present study was to isolate and identify a strain of Streptomyces w ...[more]

PMID: 25825640

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Project description:The Actinomycetia isolate PBR11 was isolated from the forest rhizosphere soil of Pobitora Wildlife Sanctuary (PWS), Assam, India. The isolate was identified as Streptomyces sp. with 92.91% sequence similarity to their closest type strain, Streptomyces atrovirens NRRL B-16357 DQ026672. The strain demonstrated significant antimicrobial activity against 19 test pathogens, including multidrug-resistant (MDR) clinical isolates and dermatophytes. Phenol, 2,5-bis(1,1-dimethylethyl), is the major chemical compound detected by gas chromatography-mass spectrometry in the ethyl acetate extract of PBR11 (EtAc-PBR11). The presence of the PKS type II gene (type II polyketide synthases) and chitinase gene suggested that it has been involved in the production of antimicrobial compounds. Metabolic profiling of the EtAc-PBR11 was performed by thin-layer chromatography and flash chromatography resulted in the extraction of two bioactive fractions, namely, PBR11Fr-1 and PBR11Fr-2. Liquid chromatography-tandem mass spectrometry analysis of both the fractions demonstrated the presence of significant antimicrobial compounds, including ethambutol. This is the first report on the detection of antituberculosis drug in the bioactive fractions of Streptomyces sp. PBR11. EtAc-PBR11 and PBR11Fr-1 showed the lowest MIC values (>0.097 and >0.048 μg/mL, respectively) against Candida albicans MTCC 227, whereas they showed the highest MIC values (>0.390 and >0.195 μg/mL, respectively) against Escherichia coli ATCC BAA-2469. The effects of PBR11Fr-1 were investigated on the pathogens by using a scanning electron microscope. The results indicated major morphological alterations in the cytoplasmic membrane. PBR11Fr-1 exhibited low cytotoxicity on normal hepatocyte cell line (CC-1) and the percent cell viability started to decline as the concentration increased from 50 μg/mL (87.07% ± 3.22%) to 100 μg/mL (81.26% ± 2.99%). IMPORTANCE Novel antibiotic breakthroughs are urgently required to combat antimicrobial resistance. Actinomycetia are the principal producers of antibiotics. The present study demonstrated the broad-spectrum antimicrobial potential of an Actinomycetia strain Streptomyces sp. strain PBR11 isolated from the PWS of Assam, India, which represents diverse, poorly screened habitats for novel microorganisms. The strain displayed 92.4% sequence similarity with genes of the closest type strain, indicating that the strain may represent a novel taxon within the phylum Actinomycetota. The metabolomics studies of EtAc-PBR11 revealed structurally diverse antimicrobial agents, including the detection of the antituberculosis drug ethambutol, in the bioactive fraction of Streptomyces sp. PBR11 for the first time. The PBR11 strain also yielded positive results for the antibiotic synthesis gene and the chitinase gene, both of which are responsible for broad-spectrum antimicrobial activity. This suggests that the untouched forest ecosystems have a tremendous potential to harbor potent actinomycetia for future drug discovery.

Project description:BackgroundKarnataka State continues to have the highest rates of maternal mortality in south India at 144/100,000 live births, but lower than the national estimates of 190-220/100,000 live births. Various barriers exist to timely and appropriate utilization of services during pregnancy, childbirth and postpartum. This study aimed to describe the patterns and determinants of routine and emergency maternal health care utilization in rural Karnataka State, India.MethodsThis study was conducted in Karnataka in 2012-2013. Purposive sampling was used to convene twenty three focus groups and twelve individual interviews with community and health system representatives: Auxiliary Nurse Midwives and Staff Nurses, Accredited Social Health Activists, community leaders, male decision-makers, female decision-makers, women of reproductive age, medical officers, private health care providers, senior health administrators, District health officers, and obstetricians. Local researchers familiar with the setting and language conducted all focus groups and interviews, these researchers were not known to community participants. All discussions were audio recorded, transcribed, and translated to English for analysis. A thematic analysis approach was taken utilizing an a priori thematic framework as well as inductive identification of themes.ResultsMost women in the focus groups reported regular antenatal care attendance, for an average of four visits, and more often for high-risk pregnancies. Antenatal care was typically delivered at the periphery by non-specialised providers. Participants reported that sought was care women experienced danger signs of complications. Postpartum care was reportedly rare, and mainly sought for the purpose of neonatal care. Factors that influenced women's care-seeking included their limited autonomy, poor access to and funding for transport for non-emergent conditions, perceived poor quality of health care facilities, and the costs of care.ConclusionsRural south Indian communities reported regular use of health care services during pregnancy and for delivery. Uptake of maternity care services was attributed to new government programmes and increased availability of maternity services; nevertheless, some women delayed disclosure of pregnancy and first antenatal visit. Community-based initiatives should be enhanced to encourage early disclosure of pregnancies and to provide the community information regarding the importance of facility-based care. Health facility infrastructure in rural Karnataka should also be enhanced to ensure a consistent power supply and improved cleanliness on the wards.Trial registrationNCT01911494.

Project description:BackgroundMaternal deaths have been attributed in large part to delays in recognition of illness, timely transport to facility, and timely treatment once there. As community perceptions of pregnancy and their complications are critical to averting maternal morbidity and mortality, this study sought to contribute to the literature and explore community-based understandings of pre-eclampsia and eclampsia.MethodsThe study was conducted in rural Karnataka State, India, in 2012-2013. Fourteen focus groups were held with the following community stakeholders: three with community leaders (n = 27), two with male decision-makers (n = 19), three with female decision-makers (n = 41), and six with reproductive age women (n = 132). Focus groups were facilitated by local researchers with clinical and research expertise. Discussions were audio-recorded, transcribed verbatim and translated to English for thematic analysis using NVivo 10.ResultsTerminology exists in the local language (Kannada) to describe convulsions and hypertension, but there were no terms that are specific to pregnancy. Community participants perceived stress, tension and poor diet to be precipitants of hypertension in pregnancy. Seizures in pregnancy were thought to be brought on by anaemia, poor medical adherence, lack of tetanus toxoid immunization, and exposure in pregnancy to fire or water. Sweating, fatigue, dizziness-unsteadiness, swelling, and irritability were perceived to be signs of hypertension, which was recognized to have the potential to lead to eclampsia or death. Home remedies, such as providing the smell of onion, placing an iron object in the hands, or squeezing the fingers and toes, were all used regularly to treat seizures prior to accessing facility-based care although transport is not delayed.ConclusionsIt is evident that 'pre-eclampsia' and 'eclampsia' are not well-known; instead hypertension and seizures are perceived as conditions that may occur during or outside pregnancy. Improving community knowledge about, and modifying attitudes towards, hypertension in pregnancy and its complications (including eclampsia) has the potential to address community-based delays in disease recognition and delays in treatment that contribute to maternal and perinatal morbidity and mortality. Advocacy and educational initiatives should be designed to target knowledge gaps and potentially harmful practices, and respond to cultural understandings of disease.Trial registrationNCT01911494.

Project description:In India, the hypertensive disorders of pregnancy and postpartum haemorrhage are responsible for nearly 40 % of all maternal deaths. Most of these deaths occur in primary health settings which frequently lack essential equipment and medication, are understaffed, and have limited or no access to specialist care. Community health care workers are regarded as essential providers of basic maternity care; and the quality of care they provide is dependent on the level of knowledge and skills they possess. However, there is limited research regarding their ability to manage pregnancy complications. This study aims to describe the current state of knowledge regarding pre-eclampsia and eclampsia among community health care workers (auxiliary nurse midwives, accredited social health activists, staff nurses) in northern Karnataka, India. Furthermore, this study describes the treatment approaches used by various cadres of community health workers for these conditions. The findings of this study can help plan focussed training sessions to build upon their strengths and to address the identified gaps.Data were collected as part of a larger study aimed at assessing the feasibility of community-based treatment for pre-eclampsia. Eight focus group discussions were conducted in 2012-2013 in northern Karnataka State: four with staff nurses and auxiliary nurse midwives and four with accredited social health activists. In addition, twelve auxiliary nurse midwives and staff nurses completed questionnaires to explore their competence and self-efficacy in managing pre-eclampsia. Qualitative data were audio-recorded, transcribed verbatim and translated for thematic analysis using NVivo 10.Community health workers described their understanding of the origins of hypertension and seizures in pregnancy. Psychological explanations of hypertension were most commonly reported: stress, tension, and fear. The most common explanation for eclampsia was not receiving a tetanus vaccination. Despite some common misperceptions regarding aetiology, these community health workers demonstrated a good grasp of the potential consequences of hypertension in pregnancy. According to auxiliary nurse midwives and staff nurses, if hypertension was detected they encouraged rest, decreased salt intake, iron supplementation and tetanus vaccination. In addition, some staff nurses administered antihypertensives, MgSO4, or other anticonvulsants. All auxiliary nurse midwives had some awareness of MgSO4, but none had administered it themselves.This study showed that knowledge regarding the aetiology of pre-eclampsia was limited. Nevertheless, their basic knowledge and skills could be strengthened to more effectively manage the hypertensive disorders of pregnancy in their communities.NCT01911494.

Project description:A total of 198 cereal samples (53 maize, 54 sorghum, 37 paddy and 54 wheat) were collected from 11 districts of Karnataka to understand the percent infection (PI), relative density (RD) and their frequency (Fr) caused by Fusarium spp. All samples were screened by agar plating method using MGA 2.5 agar media and incubated at 25 ± 2 °C for 3-5 days. The study revealed the association of 10 different Fusarium species known trichothecene producers such as Fusarium acuminatum, F. avenaceum, F. crookwellense, F. culmorum, F. equiseti, F. graminearum, F. nivale, F. poae, F. sambucinum and F. sporotrichioides along with non-trichothecene producers like F. anthophilum, F. oxysporaum, F. proliferatum, F. semitectum, F. solani, and F. verticillioides. All the ten isolated potential trichothecene producing Fusarium species were analyzed for their ability to produce trichothecenes by using thin layer chromatography method. The highest infection of Fusarium spp. in maize was by F. verticillioides with PI of (2.95 %), with RD of (15.16 %) and highest Fr was by F. graminearum (79.24 %) and the lowest was F. avenasium with PI of (0.13 %). For sorghum maximum PI was by F. verticillioides (3.02 %), with F. graminearum having highest RD (14.39 %) and with F. verticillioides highest Fr. (72.22 %). In paddy highest PI was by F. verticillioides (3.21 %) and the least was by F. avenaceum (0.09 %). Similarly in wheat the highest PI was by F. verticillioides (2.76 %) while lowest was by F. avenaceum (0.10 %). The highest Fr was with F. graminearum (79.62 %) while the lowest was by F. avenaceum (3.70 %) and the highest RD was by F. graminearum (22.04 %) and lowest was by F. solani (0.72 %). The manually identified Fusarium spp. were further confirmed by PCR-based detection using ITS1 and ITS4 primers followed by sequencing of the PCR amplicons. PCR studies confirmed that all the tested fungal isolates belongs to Fusarium spp. with the amplicon size of 600 bp. Sequencing and the blast data from NCBI data base confirmed the sequence similarity of 99 % to the genus Fusarium and accession numbers were obtained. Chemotyping studies showed that the isolated Fusarium species are known to produce different types of trichothecenes. The study revealed the diversity in phytopathogenic Fusarium spp. in major cereal crops growing in different agro-climatic regions of Karnataka, India.

Dataset Information

Isolation, Identification and Evaluation of Antimicrobial Activity of Streptomyces flavogriseus, strain ACTK2 from Soil Sample of Kodagu, Karnataka State (India).

Publications

Isolation, Identification and Evaluation of Antimicrobial Activity of Streptomyces flavogriseus, strain ACTK2 from Soil Sample of Kodagu, Karnataka State (India).

Similar Datasets

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