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Isolation and characterization of antibiotic producing microorganisms from soil samples of certain area of Punjab region of India
Abstract
Soil samples containing antibiotic producing organism are commonly employed for the production of suitable antibiotics. Isolated antibiotic may be bactericidal or bacteriostatic in nature. In the present investigation, antibiotics producing microorganisms were isolated from soil and tested against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Mycobacterium smegtitis, Proteus vulgris and Bacillus subtilis. Disc diffusion method, agar well method, streak agar method and biochemical methods have been employed to investigate the antibiotics producing microorganisms. Out of twelve isolates, two isolates; No. 5 and No. 6 were found suitable in inhibition of the growth of S. aureus and P. vulgris and zone of inhibition were found to be in ranging between 11 and 12mm respectively. The present work suggests that soil isolates, having antibiotic producing properties can be utilized commercially after proper standardization. © 2014 International Journal of Pharmaceutical and Clinical Research. All rights reserved.
... The rhizosphere microbes exhibit high level of antagonistic activity [16]. Many scientists have chosen soil for the isolation of novel antibiotics as it is a source of many antibiotic producing bacteria including Actinomycetes [13,15,[17][18][19], Constancias et al. [20] also reveals that the heterogeneity of soil results in a wide variety of ecological niches and a high diversity of soil microorganisms. This outcome correlated with our sample collection methodology implemented where the samples collected from different locations and diverse cultivations. ...
... Morphological characterization was done by Grams staining method, which is a conventional method of characterization followed by many scientists [19,22,23]. The Gram's staining indicates that all the bacterial isolates as gram positive. ...
... The isolates were screened for the production of antibiotics through primary screening (perpendicular streaking and seed overlay method) followed by secondary screening (agar well diffusion method). This is an agreement with some previous literatures which used the same methods for the screening of isolates [15,[17][18][19]24]. Bacterial culture filtrate was used in agar well diffusion method for secondary screening. ...
The present study, deal about the antibiosis activity of soil bacteria, isolated from 10 different locations of rhizosphere and diverse cultivation at Kochi, Kerala, India. The bacteria were isolated by standard serial dilution plate techniques. Morphological characterization of the isolate was done by Gram's staining and found that all of them gram positive. Isolated bacteria were tested against 6 human pathogens viz., Escherichia coli, Enterococcus sp., Pseudomonas aeruginosa, Klebsiella pneumoniae, Staphylococcus aureus and Acinetobacter sp. Primary screening was carried out by perpendicular streaking and seed overlay method. Based on the result of primary screening most potential isolates of S1A1 and S7A3 were selected for secondary screening. Both the isolates showed positive results against Enterococcus sp. and S.aureus. The maximum antagonistic activity of 20.98 and 27.08 mm zone of inhibition was recorded at S1A1 against Enterococcus sp. and S. aureus respectively, at 180 µl concentration. Molecular identification was carried out by 16S rRNA sequence. The 16S rRNA was amplified from the DNA samples by using PCR. The amplified 16S rRNA PCR products were purified and sequenced. The sequences were subjected to NCBI BLAST. The isolates S1A1 and S7A3 BLAST results showed 99% and 95% respectively, similarity with the available database sequence of Bacillus amyloliquefaciens. The sequences were deposited in GenBank and the accession numbers KY864390 (S1A1) and KY880975 (S7A3) were obtained.
... Total world production of antibiotics is more than one million tons per annum. Due to continuous endeavor made in this field, the antibiotics discovered at present are about 5,500 [1]. With the increasing number of drug-resistant pathogens, particularly the acquired multidrug resistant strains, and serious public health problems have arisen worldwide. ...
... Each 1 g of the soil sample was suspended in 10 ml of phosphate buffer saline and shake vigorously for 2-3 minutes and from the stock solution 1 ml was used to prepare the final volume of 10 3 -10 7 , by serial dilution method. This was done entirely in sterile conditions [1]. ...
... The streaks were done; the plates and slants were kept for incubation at 37°C for 24 hrs. And preserved them by keeping into the refrigerator at 4°C and maintained for a longer period by serial subculturing [1]. ...
Objective: The aim of the study was conducted to screen and isolate potential antibiotic producing bacteria from saprophytic soils collected from Potheri and Nandiambakkam. Methods: Soil was collected aseptically and subjected to serial dilution. Crowded plate technique was used for the isolation of the colony. Totally seven isolates were isolated and were screened for their antibacterial activity. The three isolates (S2A, S2B, and S3A) having better zone of inhibition were selected for morphological, microscopical, and biochemical test to prove their validity. The selected isolates were partially purified. The partially purified samples further screened for antibacterial activity, minimum inhibitory concentration (MIC) and the isolates, which shown good zone of inhibition were subjected to 16S rRNA sequencing studies to determine the species. Results: The isolates screened based on size of the zone formed. Best isolate selected by zone of inhibition was subjected to antibacterial activity, morphological, microscopical, and biochemical test, partial purification of three isolates and further screened for antibacterial and MIC. The isolate showed good zone of inhibition compared to others by MIC was selected for 16S rRNA sequencing studies. Genomic DNA extracted from isolate S2B conforms it belongs to Pseudomonas species which is named as Pseudomonas putida 2435. Conclusion: The research work revealed that the three isolates showed good antibacterial activity against Gram-positive and Gram-negative bacteria. The S2B isolate was confirmed to P. putida 2435 by 16S rRNA studies.
... Given that numerous dynamic systems influence the growth inhibition zone's size and location. According to Kaur et al, (2020), comparing antibiotic agents based solely on the extent of their inhibition zones is dubious. ...
... The presence of carbonic acid, glycerin, nonadecane, 5hydroxymethyl furfural, L-arabinitol, methyl chloroformate, and other derivatives was shown by the GC-MS analysis, which was carried out to determine the bioactive chemicals of the isolated bacterial extracts. According to earlier research, the aforementioned substances have antibacterial properties (Kaur et al., 2020). The chemical formulas and structures of some significant substances included in bacterial samples were shown in Table 6. ...
Antibiotics produced by soil microorganisms, are crucial for treating bacterial and fungal diseases. Due to antibiotic resistance, a broad spectrum of antibiotics from bacteria is needed. Soil samples from Zamfara state were used for detecting antibiotic-producing bacteria. The study involved physico-chemical analysis, pour plate methods, biochemical tests and molecular characterization using 16s rRNA. Agar well diffusion method was employed to test isolates’ ability to inhibit pathogens growth and their potential for antibiotic production. Antibiotics production was assessed through small-scale submerged fermentation condition. Compounds were extracted using solvent extraction method, thin layer chromatography and GC-MS analysis were used for separation and identification. Data analysis were done using SPSS and were presented as mean, standard error of mean, and Anova. The soil samples exhibited light brown to dark brown colors, indicating clay-loamy, loamy-sandy and sandy-loamy. Sample C had the highest pH (7.31±.03), Sample H had the highest temperature (26oC), sample D had the highest electrical conductivity (108µs/m), and sample B had the highest moisture content (5.6). Six bacteria were identified and these include; Bacillus cereus, Bacillus pumilus, Providencia stuartii, Providencia alcalificiens, Pseudomonas aeruginosa, and Micrococcus luteus. All the organisms inhibited at least one pathogen, with Bacillus cereus showing the highest zone of inhibition. GC-MS analysis revealed diverse bioactive compounds namely, Carbonic acid, Glycerin, Nonadecane, 5-hydroxymethyl furfural, L-arabinitol, and Methylchloroformate. This study has demonstrated the diversity of bacterial population in the local soils and potential production of antibiotics that can be useful for many medical applications and further studies are recommended.
... An endophytic fungus Taxomyces andreanae is able manufacture produce paclitaxel [11]. In turn, paclitaxel helps to prevent breast and pancreatic cancers [13]. Camptothecin is another antibiotic synthesized from endophytic fungi, including Aspergillus sp. and Trichoderma atroviride [7]. ...
... The antibiotic is used for the treatment of severe bladder toxicity, ovarian cancer and colorectal cancer, respectively [6]. Vinblastine and vincristine are anticancer drugs secreted from Catharanthus roseus, Alternaria, Fusarium oxysporium [13], aryltetralin-type lignan and podophyllotoxin are synthesized by Phialocephela fortinii [12]. The drugs are used for the treatment of anticancer disease [14][15][16][17][18][19], (Table 1). ...
... Nike et al. [36] and Kaur et al. [37] have used the primary and secondary screening methods as done in our work for the isolation and screening of bacterial isolates. Many scientists have adopted the agar well diffusion method for secondary screening of bacteria using cellfree culture filtrates [38][39][40]. ...
... Morphological and biochemical characterization of isolates PR1, PR2, and PR3 were carried out as it is a tool for preliminary identification of bacteria, and it is a conventional method followed by microbiologists all over the world [37,60]. For the past several decades, most laboratories adopt microscopic identification and biochemical characterization to identify the bacteria [61]. ...
Background
The present study dealt with the screening of soil bacteria with antibacterial activity from different locations in Bangalore, India. Antibiotics play the role of self-defense mechanism for the bacteria and are produced as secondary metabolites to protect themselves from other competitive microorganisms. The need for new antibiotics arose as the pathogenic bacteria acquire resistance to various antibiotics meant for treating human diseases. Given the importance of antibiotics of bacterial origin, standard techniques have been used to isolate and characterize the soil bacteria which showed antibacterial activity.
Results
The isolated bacteria were tested against human pathogenic bacteria like Staphylococcus aureus , Escherichia coli , Pseudomonas aeruginosa , and Klebsiella pneumoniae by primary and secondary screening methods. The isolates PR1, PR2, and PR3 were confirmed to have antibacterial activity against S. aureus , E. coli , P. aeruginosa , and K. pneumoniae by both methods. Studies on the effect of filter sterilization, autoclaving, and proteinase K treatment on culture filtrates showed filter sterilization as the best method. The effect of different carbon and nitrogen sources on the antibacterial activity showed that preference by each isolate differed for carbon and nitrogen requirements. The isolates PR1, PR2, and PR3 were identified as Bacillus aryabhattai strain PR-D07, Arthrobacter humicola strain PR-F07, and Neomicrococcus lactis strain PR-F11 through 16S rRNA sequencing.
Conclusion
Findings from this research work are encouraging and could proceed further to applied aspects. Only 3 bacterial isolates out of 263 isolates from soil samples displayed antibacterial activity against human pathogens S. aureus , E. coli , P. aeruginosa , and K. pneumoniae . They were identified as B. aryabhattai , A. humicola , and N. lactis by 16S rRNA studies and all of them are Gram-positive. Each isolate preferred different carbon and nitrogen sources for their enhanced antibacterial activity. Efficacy of the culture filtrates of these isolates was tested by filter sterilization, autoclaving, and proteinase K treatment. Filter-sterilized culture filtrates showed higher antibacterial activity than other treatments. A comparison of the antibacterial activity of culture filtrates and antibiotic streptomycin produced an inhibition zone of 18.5 mm and 15.5 mm respectively. This is the first report on the antibacterial activity of all the 3 bacterial strains ( B. aryabhattai strain PR-D07, A. humicola strain PR-F07, and N. lactis strain PR-F11), against all the human pathogens, mentioned earlier. It is also found that the antibiotic factor is proteinaceous as proteinase K considerably reduced the antibacterial activity of the culture filtrates. With the above significant results, these 3 bacteria are considered to be promising candidates for the isolation of new antibacterial agents.
... Antibiotik tersebut dapat bersifat sebagai bakteriostatik di alam. Mikroorganisme yang memiliki kapasitas lebih dalam memproduksi antibiotik umumnya dapat bertahan hidup lebih lama dibandingkan yang memproduksi antibiotik dalam jumlah yang lebih sedikit(Kaur et al, 2014). Berbagai penelitian telah menunjukkan bawa mikroorganisme yang diisolasi dari limbah atau tempat kotor dapat menghasilkan zat antimikroba. ...
... Berbagai penelitian telah menunjukkan bawa mikroorganisme yang diisolasi dari limbah atau tempat kotor dapat menghasilkan zat antimikroba. Penelitian olehKaur et al, 2014 telah berhasil mengisolasi bakteri tanah yang mampu menghasilkan zat antimikroba dan telah diuji tehadap bakteri E. coli, Pseudomonas aeruginosa, Staphylococcus aureus, Mycobacterium smegmatis, Proteus vulgaris dan Bacillus subtilis. hasil yang didapatkan dari penelitian diketahui bahwa isolasi dan uji potensi secara in vitro pada talenanpotongikan terdapat berbagai macam jenis bakteri yang berpotensi untuk menghambat pertumbuhan bakteri sehingga berpotensi untuk dijadikan sebagai sumber antibiotic terhadap infeksi bakteri Salmonella sp. ...
... Antibiotik tersebut dapat bersifat sebagai bakteriostatik di alam. Mikroorganisme yang memiliki kapasitas lebih dalam memproduksi antibiotik umumnya dapat bertahan hidup lebih lama dibandingkan yang memproduksi antibiotik dalam jumlah yang lebih sedikit(Kaur et al, 2014). Berbagai penelitian telah menunjukkan bawa mikroorganisme yang diisolasi dari limbah atau tempat kotor dapat menghasilkan zat antimikroba. ...
... Berbagai penelitian telah menunjukkan bawa mikroorganisme yang diisolasi dari limbah atau tempat kotor dapat menghasilkan zat antimikroba. Penelitian olehKaur et al, 2014 telah berhasil mengisolasi bakteri tanah yang mampu menghasilkan zat antimikroba dan telah diuji tehadap bakteri E. coli, Pseudomonas aeruginosa, Staphylococcus aureus, Mycobacterium smegmatis, Proteus vulgaris dan Bacillus subtilis. hasil yang didapatkan dari penelitian diketahui bahwa isolasi dan uji potensi secara in vitro pada talenanpotongikan terdapat berbagai macam jenis bakteri yang berpotensi untuk menghambat pertumbuhan bakteri sehingga berpotensi untuk dijadikan sebagai sumber antibiotic terhadap infeksi bakteri Salmonella sp. ...
Dendrobium is a member of Orchidaceae. This genus is the third largest genus only after Bulbophyllum and Epidendrum, reaches 1500 species. Bogor Botanical Gardens has 500 species of orchids with 104 of them are Dendrobium. A total of 133 specimens have not been identified because morphologically, they have commonly low variations among species or even have a high degree of plasticity. The approach through DNA barcoding is expected to help species identification effectively. This research was conducted as a first step in the process of selecting the best DNA barcoding primer for species identification in Dendrobium. The DNA Barcoding sequence data namely ITS2, rbcL, matK, and psbA-trnH sequences were analyzed using MEGA X and constructed into dendrograms. Unfortunately, the trees generated from rbcL, psbA-trnH, and ITS2 showed a quite clear determination yet some species are clustered in different subgenus clad. On the other hand, the matK tree produce a very clear species determination and can be recommended to be used as a single DNA Barcoding primer for identifying the Dendrobium sp. in Bogor Botanic Gardens collection.
... Antibiotik tersebut dapat bersifat sebagai bakteriostatik di alam. Mikroorganisme yang memiliki kapasitas lebih dalam memproduksi antibiotik umumnya dapat bertahan hidup lebih lama dibandingkan yang memproduksi antibiotik dalam jumlah yang lebih sedikit(Kaur et al, 2014). Berbagai penelitian telah menunjukkan bawa mikroorganisme yang diisolasi dari limbah atau tempat kotor dapat menghasilkan zat antimikroba. ...
... Berbagai penelitian telah menunjukkan bawa mikroorganisme yang diisolasi dari limbah atau tempat kotor dapat menghasilkan zat antimikroba. Penelitian olehKaur et al, 2014 telah berhasil mengisolasi bakteri tanah yang mampu menghasilkan zat antimikroba dan telah diuji tehadap bakteri E. coli, Pseudomonas aeruginosa, Staphylococcus aureus, Mycobacterium smegmatis, Proteus vulgaris dan Bacillus subtilis. hasil yang didapatkan dari penelitian diketahui bahwa isolasi dan uji potensi secara in vitro pada talenanpotongikan terdapat berbagai macam jenis bakteri yang berpotensi untuk menghambat pertumbuhan bakteri sehingga berpotensi untuk dijadikan sebagai sumber antibiotic terhadap infeksi bakteri Salmonella sp. ...
Phyllanthus merupakan satu dari 58 marga anggota suku Phyllanthaceae. Marga ini memiliki jumlah anggota terbesar yakni mencapai 1200 jenis dan sebanyak 32 jenis dapat ditemukan di Indonesia. Identifikasi jenis pada marga ini cukup sulit untuk dilakukan karena beberapa jenis memiliki ciri yang serupa. Penggunaan identifikasi dini menggunakan DNA Barcoding diharapkan mampu membantu kegiatan identifikasi jenis secara lebih cepat dan efektif. Penelitian ini dilakukan untuk mengetahui karakter dan membandingkan primer DNA Barcoding yang baik untuk digunakan dalam identifikasi jenis-jenis Phyllanthus. Data sekuen DNA Barcoding yang berasal dari sekuen inti dan plastida diperoleh melalui data mining dari NCBI. Konstruksi dendrogram dilakukan menggunakan metode UPGMA berdasarkan parameter Kimura-2 melalui software MEGA X. keempat dendrogram yang dihasilkan menunjukkan resolusi yang cukup baik dalam mendeterminasi jenis-jenis Phyllanthus, tetapi sekuen ITS dan matK menghasilkan dendrogram dengan determinasi lebih baik karena pengelompokan jenisnya bersesuaian dengan pengelompokan anak marganya.
... In order to isolate and identify root-associated bacteria with a potential to inhibit the test strain organisms (E. coli, B. subtilis and S. aureus), isolation was undertaken using serial dilution, pour plate method, purification and broth culturing, preparation of cell free supernatant 18 followed by antibiotic sensitivity test 19 and molecular identification of 16S rRNA gene sequencing. 20 ...
... T0(negative control) T1, T2, T3, T5, T6 and T7 did not inhibit B. subtilis with a mean diameter of zero(0 mm). 18 The significantly higher potential of T4 to inhibit B. subtilis compared to the positive control (Tetracycline) signifies a potentially effective antimicrobial compound/peptide produced by the root-associated bacteria (T4). ...
The study was carried out to isolate and identify potential antibiotic-producing bacteria associated with water cabbage (Pistia stratiotes) roots collected from Pampanga River, Pampanga, Philippines. Seven (7) bacterial colonies were randomly chosen at the 10-6 dilution factor. Antibiotic sensitivity test using agar well method revealed that only one isolate out of 7 selected colonies can inhibit the growth of the test organisms. Specifically, the isolate (namely T4) supernatant inhibited E. coli and B. subtilis but not S. aureus. T4’s ability to inhibit E. coli was comparable with that of Tetracycline (positive control). Surprisingly, its inhibition of B. subtilis is significantly higher than that of Tetracycline. 16S rRNA gene sequence analysis using NCBI Basic Local Alignment Search Tool revealed 99% similarity of the isolate (T4) with Exiguobacterium acetylicum, a gram-positive, antibiotic-producing bacterium previously isolated from an apple orchard rhizosphere.
... At low physiological amounts, its role in cell signaling is significant (Fu et al. 2018). H 2 S is a gas that regulates major functions in the nervous, cardiovascular, immune, and gastrointestinal systems of higher organisms (Kaur et al. 2014). ...
Antimicrobial resistance in bacteria is a global threat that can make antibacterial treatments ineffective. One well-known method of antibiotic resistance and a common defensive mechanism in many harmful bacteria is the synthesis of endogenous hydrogen sulfide (H2S) in bacteria. In this study, soil bacteria were screened using the lead acetate agar test and the triple sugar iron test to determine that they were non-endogenous H2S producers. This was further validated by full genome analysis of the identified organism against the gene sequences of H2S-producing genes. Antibacterial resistance of the bacteria was phenotypically analyzed using the Kirby-Bauer disk diffusion method. Then, the effect of exogenous H2S on the antibiotic-resistant bacteria was checked in sodium sulfide, leading to antibiotic re-sensitization.
... A conventional technique for gathering samples, the random sampling method was outlined by Williams and Vickers (1986); this process lends support to the current investigation. A conventional technique recommended by multiple scientists, Gram's staining was used to identify the morphological characteristics (Diddi et al., 2013;Kaur et al., 2014;Kumar et al., 2016). Hence, in order to gain a deeper understanding of its morphological traits, soil bacterial isolates were appropriately stained using Gram's staining method, which showed that the majority of the isolates were gram-positive. ...
... After collection, by uprooting the plant, the soil samples from the root were kept in a sterile, labelled, airtight zip-lock cover and refrigerated at 4 °C for further analysis [18]. Plant specimens were identified according to the herbarium guidelines [19]. ...
Background
Over the previous two decades, Plasmodium falciparum strains have become increasingly resistant to several medications. As a result, there is an urgent need to develop new therapeutic options. Taking this into account, we focused our research on screening microbial extracts from rhizosphere soil samples in specific regions, which increases the likelihood of discovering bacteria capable of producing antiplasmodial activity.
Results
In the current study, we aimed to isolate thirty-two different medicinal plant rhizosphere soil samples collected from Kolli Hills (January–December 2016). Isolation was performed on nutrient and starch casein agar medium by serial dilutions, and distinct colonies were chosen from each dilution. A total of two seventy-five bacterial isolates were isolated from the research plants and kept as pure cultures on nutrient agar. In which, maximum count of fourteen Gram-positive spore forming bacilli strains have been identified and further evaluated for morphological, cultural, and biochemical traits and significantly identified as Bacillus species. Further, promising anti-plasmodial action was demonstrated by B. megaterium bacterial extracts, with IC 50 values of 24.65 µg/mL at 24 h and 7.82 µg/mL at 48 h. Bacillus mycoides showed good antiplasmodial activity with (IC 50 P. falciparum 3D7: 23.52 μg/mL at 24 h and 22.88 μg/mL at 48 h, Bacillus flexus showed IC 50 of 18.36 and 6.24 μg/mL and moderate antiplasmodial activity observed in Bacillus tequilensis. Poor antiplasmodial activity was found in Bacillus subtillis , Bacillus macerans, Bacillus pumilus and Bacillus larvey . Interestingly, 16S rRNA sequencing results confirmed that our bacterial species was Bacillus megaterium with 99% similarity observed with the accession number KX495303.1. Additionally, GC–MS analysis revealed effective anti-plasmodial bioactive compounds.
Conclusions
These findings show the potential of B. megaterium from Achyranthes aspera as a antiplasmodial agent. However, more research is needed to fully understand the bioactive compound of these strains and further studies are necessary to explore drug formulation and toxicity levels in the future.
... One gram (1g) of each soil samples was suspended in 9mL of sterile distilled water and vortex for 2-3 minutes. From the mother solution (stock), 1mL of the supernatant was aseptically pipetted and transferred into the next tube and serial dilution continues down to the last tube [10] [3]. ...
The investigation into soil bacteria has been widely studied and becoming increasingly appreciated as an exceptional reservoir of unique naturally occurring biologically active metabolites with pharmaceutical applications. This article aimed to isolate, identify and biochemically characterize antibiotic-producing bacteria from anthill soils in the permanent site of Ibrahim Badamasi Babangida University, Lapai (IBBUL), Niger State, Nigeria. The sum of ten samples were collected from five sampling sites, the sampling was done in threefold (morning, noon and evening) and analyzed adopting standard microbiological protocols. The obtained result revealed that the total bacteria count in the morning ranges from 2.1×107 cfu/mL to 1.4×106 cfu/mL, noon count ranges from 3.1×107 to 2.6×106 cfu/mL while evening count was in the range of 2.1×107 cfu/mL to 1.7×106 cfu/mL. A total number of five (5) bacteria were isolated as Staphylococcus aureus, Staphylococcus epidermidis, Bacillus subtilis, Bacillus lentus and Micrococcus reseus. The total prevalence of the bacterial isolates in the morning, noon and evening were calculated as B. subtilis (109.08%), S. epidermidis (36.36%), M. reseus (36.36%), B. lentus (63.63%), and S. aureus (54.54%) respectively. These isolates were further assayed against Escherichia coli, Salmonella typhi, Klebsiella sp. and Staphylococcus aureus. The antibacterial outcome showed that two (2) (40%) anthill isolates exhibited antibacterial activity against three (3) tested bacteria (Escherichia coli, Salmonella typhi and Staphylococcus aureus). This research study has showcased that the production of inhibitory substances are common among some of the bacterial strains isolated from anthills.
... In the current study, some bacteria isolates (Streptomyces lydicus and Bacillus subtilis) and fungal isolates (Penicillium digitatum, Aspergillus niger and Aspergillus flavus), identified from the TMs are often used for the synthesis of antibiotics on industrial scale. 38 Similar report showed presence of Bacillus subtilis in soil consumed by people of Cameroon and South Africa. 39 ...
The search for novel antibiotics for treating human-wildlife infections has not stopped. This study isolated and characterized bacteria and fungi from wildlife geophagic termite mounds (TMs) using standard microbiological procedure. Four composite samples of TMs evident to be eaten by wildlife were randomly collected (TM1, TM2, TM3 and TM4) in addition with a composite forest soil (FS5) that showed no sign of consumption. With the use of appropriate media, all samples were subjected to microbiological analysis covering morphological, biochemical (for bacteria), colonial and cellular morphology (for fungi). Bacteria and fungi cells isolated and characterized from respective soil samples include; Streptomyces lydicus/Aspegillus niger (TM1), Paenibacillus polymyxa/Penicillium digitatum (TM2), Pseudomonas fluorescens/Fusarium spp. (TM3), Bacillus subtilis/Aspergillus flavus (TM4) and Acinetobacter radioresistens/Geotricihum candidum (FS5), all of which have both beneficial and harmful characteristics. Wildlife may be affected by pathogenic organisms when consuming TMs for medicinal reasons. Dhaka Univ. J. Pharm. Sci. 21(1): 53-58, 2022 (June)
... In order to identify the morphological attributes Grams staining procedure was followed, which is a traditional method conferred by several scientists. [90][91][92] Therefore, further comprehension of its morphological characteristics appropriate staining of soil bacterial isolates with Gram's staining method showed that mostly bacterial isolates ascertained gram positive. However, outcomes regarded as confirmatory for Wadetwar and Patil, 87 stated that most of the soil microbiome after testing found to be gram positive. ...
The contamination of environmental sully with antibiotics is regarded as a major problem today and predictable to attain more recognition in near future. However, human intervention resulting in antibiotic consumption is being enhancing all around the world. Our review of literature revealed the role of microbiome in sully and how antibiotic resistant genes raised. The structure of antibiotics basically influenced by natural components such as biotic and abiotic push which shifts based on different soils. Therefore, management of microbiome in soil and their expression studies were distinctively revealed. The assessment of antibiotic resistance genes with help of next generation sequencing provided a clear comprehension on genome and transcriptome of the bacterial genes. Thus, interaction of microbiome with soil can also be well understood. The current findings in our study will guide every researcher to follow logical protocol in analyzing microbiota composition is covered as well and also to understand its metagenomic and sequenced with next-generation sequencer which helps to comprehend the diverse micro-flora present in soil and its operation. Finally, later progresses in bioinformatics computer program, flow of work, and applications for analyzing metagenomic information are put in a nutshell.
... The method of Kaur et al. (2014) was used in preparing the samples. Firstly, the wet weight of the first samples contained within the core sampler were measured using weighing balance and recorded. ...
The emergence of new diseases and multiple-antibiotic resistance pathogens that resist the action of clinically used antibiotics have increased the needs of discovering new antibiotics. However, soil served as the most important target for most researchers in their attempt of discovering new antibiotics that have pharmaceutical values. The aim of this study was to detect the antibiotic-producing bacteria from soil samples in parts of Wudil local government of Kano state using standard procedure. The color ranged from light brown to dark brown. The texture ranged from clay loam, clay and loam. Sample C was found to have the highest mean pH and electrical conductivity of 7.46 and 108.90μs/cm, respectively, while sample B had the least mean values of 6.95 and 73.53μs/cm, respectively. The highest mean bulk devsity and particle density was sample B with 1.60g/cm3 and 1.87g/cm3, respectively, while sample C had the least mean values of 1.48g/cm3 and 1.71g/cm3, respectively. Sample C was found to have the highest mean moisture content and porosity with 17.93% and 15.57%, respectively, while sample A had the least mean values with 10.61% and 10.74%, respectively. Four bacterial spp. were identified based on morphological description, gram’s reaction and biochemical tests, and were identified as Bacillus spp., Micrococcus spp., Pseudosomonas spp. and Proteus spp. Sample C was found to have the highest mean total bacterial viable plate count of 19.9×105cfu/g, while sample B had the least mean value of 4.1×105cfu/g. Bacillus spp. was found to have the highest percentage occurrence with 21(47%), while Proteus spp. had the least value with 4(9%). E. coli had the highest and least susceptibility to metabolites produced by Bacillus spp. and Pseudosomonas spp. with inhibition zones of (17 and 0mm) respectively. While the control (Erythromycin 250mg/ml) produced the inhibition zones of (27 and 21mm) against E. coli and S. aureus respectively. It was concluded that soil samples in Lajawa, Kausani, Wudil Sabon gari and Wudil Tsohon gari of Wudil local government contained the bacteria with potentiality of producing antimicrobial substance and recommended the use of molecular techniques for further characterization.
... Pharmaceutical industry all over the world is producing more than one million tons antibiotics per annum. About five thousand and five hundred antibiotics have been discovered due to constant efforts made in this field (Kaur et al., 2014). Presently several micro-organisms are grown in artificial medias for the search of antibiotic producing microorganisms. ...
Many soil microorganisms' i.e., bacteria and fungi produce secondary metabolites called antibiotics. These are used for the treatment of some of the bacterial, fungal and protozoal diseases of humans. There is a need for isolation of a broad spectrum of antibiotics from microorganisms due to the emergence of antibiotic resistance. In the present study two antibiotic producing bacteria Klebsiella pneumoniae and Bacillus cereus were isolated from pharmaceutical and poultry feed industry of Hattar, Haripur Pakistan. Total 10 waste samples were collected from different industries (Marble, Ghee, Soap, Mineral, Steel, Poultry Feed, Pharmaceutical, Qarshi, Cosmetic and Glass). Thirty-three bacterial strains were isolated from industrial wastes of these ten different industries. Fourteen out of thirty-three bacterial strains exhibited antimicrobial activities against at least one of the test microbes considered in this study including Escherchia coli, Staphylococcus aureus and Salmonella typhi. The bacteria were isolated by standard serial dilution spread plate technique. Morphological characterization of the isolates was done by Gram staining. Nine bacterial isolates out of fourteen were initially identified as B. cereus and five as K. pneumoniae through biochemical characterization. The antibacterial activities were tested by well diffusion method. Maximum number of antibiotic producing bacteria were isolated from pharmaceutical and poultry feed industry based on the results of primary screening, the most potential isolates S9, S19, S20, S22 and S23 were selected for secondary screening. The maximum activity against E. coli and S. aureus was recorded by bacterial isolate S19 i.e zones of inhibition of 6.5mm and 9mm while S20 showed 7.5mm and 6mm zones respectively. Molecular identification was carried out on the basis of 16S rRNA sequence analysis. Finally, the isolates were identified as B. cereus accession number LC538271and K. pneumoniae accession number MT078679. Analysis of bacterial extract S20 through GC-MS indicated the presence of 8 compounds of diverse nature and structure. Present study suggests that wastes of pharmaceutical and poultry feed industry may have antibiotic producing bacteria. These bacteria could be utilized for the production of antibiotics. B. cereus and K. pneumoniae isolated from wastes of poultry feed and pharmaceutical industries have the potential to produce antibiotics and could be used to control the microbial growth. Keywords: antibiotic producing microorganisms, antibacterial activity, zone of inhibition, soil sample, well diffusion method, spread plate technique. Resumo Muitos microrganismos do solo, ou seja, bactérias e fungos produzem metabólitos secundários chamados antibióticos. Eles são usados para tratamento de algumas doenças bacterianas, fúngicas e protozoárias em humanos. Há necessidade de isolamento de um amplo espectro de antibióticos de microrganismos devido ao surgimento de resistência aos antibióticos. No presente estudo, duas bactérias produtoras de antibióticos, Klebsiella pneumoniae e Bacillus cereus, foram isoladas da indústria farmacêutica e de ração avícola de Hattar, Haripur, Paquistão. Um total de 10 amostras de resíduos foi coletado de diferentes indústrias (mármore, ghee, sabão, mineral, aço, ração para aves, farmacêutica, Qarshi, cosmética e vidro). Trinta e três cepas bacterianas foram isoladas de resíduos industriais dessas dez diferentes indústrias. Quatorze das 33 cepas bacterianas exibiram atividades antimicrobianas contra
... One gram (1g) of each soil samples was suspended in 9mL of sterile distilled water and vortex for 2-3 minutes. From the mother solution (stock), 1mL of the supernatant was aseptically pipetted and transferred into the next tube and serial dilution continues down to the last tube [10] [3]. ...
The investigation into soil bacteria has been widely studied and becoming increasingly appreciated
as an exceptional reservoir of unique naturally occurring biologically active metabolites with
pharmaceutical applications. This article aimed to isolate, identify and biochemically characterize
antibiotic-producing bacteria from anthill soils in the permanent site of Ibrahim Badamasi Babangida
University, Lapai (IBBUL), Niger State, Nigeria. The sum of ten samples were collected from five
sampling sites, the sampling was done in threefold (morning, noon and evening) and analyzed
adopting standard microbiological protocols. The obtained result revealed that the total bacteria
count in the morning ranges from 2.1×107 cfu/mL to 1.4×106 cfu/mL, noon count ranges from
3.1×107 to 2.6×106 cfu/mL while evening count was in the range of 2.1×107 cfu/mL to 1.7×106
cfu/mL. A total number of five (5) bacteria were isolated as Staphylococcus aureus, Staphylococcus
epidermidis, Bacillus subtilis, Bacillus lentus and Micrococcus reseus. The total prevalence of the bacterial isolates in the morning, noon and evening were calculated as B. subtilis (109.08%), S.
epidermidis (36.36%), M. reseus (36.36%), B. lentus (63.63%), and S. aureus (54.54%)
respectively. These isolates were further assayed against Escherichia coli, Salmonella typhi,
Klebsiella sp. and Staphylococcus aureus. The antibacterial outcome showed that two (2) (40%)
anthill isolates exhibited antibacterial activity against three (3) tested bacteria (Escherichia coli,
Salmonella typhi and Staphylococcus aureus). This research study has showcased that the
production of inhibitory substances are common among some of the bacterial strains isolated from
anthills.
... The isolates were screened for antibiotics production through disc diffusion in the in vitro plate assay. This is an agreement with some previous literatures that used the same methods to screen prospective antibiotic producing isolates [26,39,40] After investigating the results, the present study strain could be used as potential antibiotic candidate after purification and necessary strain development processes. However, considering the limited resources and financial constraints, those steps were beyond our study objectives. ...
The bacterial resistance over antibiotics has gained significant concern worldwide for the last few decades. This study aimed to isolate and characterize antibiotic producing bacteria from different water sediment sources in Bangladesh. A total of 9 samples were collected from three different sediment sources from Gopalganj, Bangladesh. Bacterial isolates grown in nutrient agar with cycloheximide were tested for antagonistic activity following disc-diffusion assay. Based on preliminary careening, three isolates namely LIS01, LIS02 and LIS03 showed antibacterial activity against six clinical isolates: E. coli, Salmonella sp., Shigella sp., Klebsiella pneumoniae, Streptococcus sp., and Staphylococcus sp. The isolate LIS01 with the highest (P<0.05) zone of inhibition was characterized using 16S rRNA gene sequencing. According to phylogenetic
analysis, the isolate was identified as Exiguobacterium indicum with more than 99.7% sequence similarity to previously characterized strains. The results of our study are very promising considering the antibiotic resistant bacteria and could be helpful in treating diseases associated with drug failure caused.
... Hence, species such as Streptomyces, Bacillus and Penicillium have been researched constantly for their antibiotic production capability. Bacillus species, the predominant soil bacteria because of their resistant endospore formation and production of vital antibiotics (4) . The major antibiotics reported till date are from actinomycetes. ...
Soil being a major reservoir for microorganisms it is a source of interest for isolation of antibiotic producing organisms. The emergence of antibiotic resistance and need for better, broad spectrum antibiotics is always in high demand. In the present study, antibiotic producing bacteria were isolated from a local soil sample. Total ten soil samples were collected from local pond aseptically and subjected to serial dilution. Crowded plate technique was employed for the isolation of the colony. Total five isolated were isolated which exhibited zone of inhibition around the colony. The isolated colonies were subjected to morphological, microscopical and biochemical characterization. All five colonies were found to be gram positive, non-sporulating organisms and found they belong to the Actinobacteria class. The isolated colonies were subjected to screening for antimicrobial activity against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus subtilis and Yeast by perpendicular streak method. The primary screening results conclude that except one colony all have good antimicrobial activity. One colony found to be highly potential activity which had inhibition towards gram positive, gram negative, sporulating and fungal activity. This study may contribute in providing information on the antibiotic producing microorganisms in soil. Further characterization, purification, and structural elucidation are recommended to know the novelty, quality and commercial value of these antibiotics.
... The results revealed 3 isolates with mainly yellow-white opaque morphological characteristic for studied colonies in accordance with similar studies that suggested soil as an important natural resource of medically important bacteria 14 . In agreement with similar the resultsthe biochemical tests of the four isolates revealed that all isolated were positive for catalase production 15 . ...
Currently, there is an increase prevalence of antibiotic-resistant bacteria worldwide. Therefore, the need for characterization of naturally occuring antibiotics with less antibiotic resistance is required. Soil resources contains valuable antibiotic producing microorganisms that increasingly being utilized for the production of suitable antibiotics. Therefore, this study aimed at identifying an antibiotic bacteria with ability of producing antibiotic that is isolated from soil samples collected from Al Zarqa provenance, an arid area in Jordan. Morphological and biochemical characterization of the isolates were carried out and found that all of the isolates belong to Bacillus genus. Further confirmation of the characterization of the bacteria was done by ribosomal RNA and PCR. The results reveal that the isolates represent Basilluslicheniformis. These bacilli were further investigated for antimicrobial activities against 6 ATCC human pathogens viz., S. aureus, S. pneumonia, Salmonella typhi., E. coli, P. mirabels and E. cloacae. Additionally, the results of Gas Chromatography Mass Spectrometry (GCMS) of ethyl acetate extracts for B. licheniformis secondary metabolites showed that they contain two main antimicrobial compounds namely Pyrrolo [1, 2-a] pyrazine-1, 4-dione,hexahydro and Trans-13-octadecenoic acid. The present work maybe suggests that soil isolates from the studied arid area include antibiotic producing strains that can be utilized commercially.
... The results suggested that consumption of GT can cure a patient suffering from bacterial infections caused by S. aureus and P. vulgaris as well as fungal infections due to Cryptococcus sp. and Penicilium sp. Most of the research results have already claimed that GT has powerful antimicrobial property against V. cholera, Salmonella sp., Clostridium sp.23 , S. dysenteriae, A. fumigatus, C. albicans, S. typhimurium 1402/84, S. typhi Ty2a, S. dysenteriae, Y. enterocolitica C770 and E. coli (EPEC P2 1265) ...
Green tea (Camellia sinensis) consists of various bioactive components which is useful in new kind of formulation to treat the human diseases. Microorganisms became resistant to varieties of antibiotics so natural herbal remedy is needed to combat the microbial infections. In the present study, polyphenols such as MQK (myricetin, quercetin, kaempferol) and EGCG (epigallocatechingallate) and C (catechin) were extracted from green tea (Camellia sinensis) leaves and tested against test bacteria and fungi. Different concentrations of these polyphenols were examined for antibacterial activities by disc diffusion method. Antimicrobial assay showed significant zone of inhibition against S. aureus (Gram positive bacteria), P. vulgaris (Gram negative bacteria), Cryptococcus sp.(fungus) and Penicilium sp. (fungus). Catechin showed broad spectrum antibacterial and antifungal activities. Minimum inhibitory concentration (MIC) of catechin was 3.12 mgdl-1 against S. aureus, P. vulgaris and Cryptococcus sp. whereas at 6.25 mgdl-1 inhibited Penicilium sp. MQK and EGCG were especially effective against Cryptococcus sp and Penicilium sp.which gave MIC value of 6.25 mgdl-1. EGCG at same concentration was also effective against P. vulgaris. This study suggests the therapeutic use of green tea against bacterial as well as fungal infections.
... One gram each of the soil samples was suspended in 9ml of distilled water and vortex for 2-3 minutes to obtain homogenous mixture and carried out serial dilution. From the stock culture, a volume of 1mL was aseptically transferred to the next tube and serial dilution continues down to tube 6 [21]. ...
Antimicrobial agents or antibiotics are the most significant commercially available and utilized secondary metabolites, which are highly produced by the soil microbes (bacteria and fungi) and found to be effective and broad spectrum. Microbes produce metabolic products (antimicrobial agents) through the process called antibiosis. Majority of the classes of antibiotics used are derivatives of animals (microbes) and floras (plants). But currently, the microbial resistance is at the top gear which requires more effort to come up with novel structure, effectual, toxic free and reasonable cost of new antimicrobial products against microbial infections. In the present study, a trial was made to isolate, identify and characterize the antibiotic producing bacteria from the soil samples collected from different sites of abattoir in Lapai, using the standard microbiological techniques. A total of nine (9) bacterial of both groups (Gram positive and negative) which includes Corynebacterium diphtheriae, Bacillus cereus, Escherichia coli, Enterobacter aerogene, Clostridium specie, Bacillus subtilis, Pseudomonas species, Staphylococcus aureus and Klebsiella pneumoniae were isolated in the course of the study. Pseudomonas species was the most frequently isolated bacteria (33.33%) while the rest of the isolates were 8.33% across the five different sampling sites. These isolates were further screened against some pathogenic microbes viz, Salmonella sp, Klebsiella pneumoniae, Pseudomonas aeruginosa and Candida albicans. Three of the bacterial isolates isolated from the abattoir were found to exhibits antimicrobial activity against two pathogenic bacteria used in this study. This study indicates that some of the soil microorganisms could be an interesting source of antimicrobial bioactive substances.
... Soil is one of the best sources for isolating novel antibiotics as many scientists have chosen it for their research [13][14][15][16]. It was reported that the heterogeneity of soil environment results heterogeneous population of soil bacteria [17]. ...
Antibiotics are the secondary metabolites produced by bacteria and fungi to defend themselves from other pathogens. These secondary metabolites are being produced and used as a drug to cure different diseases. However, antibiotic resistance is a common problem that demands an urgent need to discover new antibiotics routinely. Several approaches have been performed to develop novel and potent antibiotics from natural sources against pathogenic bacteria. Among the different sources soil has been considered as the potent natural source of obtaining bacteria with the ability to produce novel antibiotics. The present work has been focused on the isolation of antibiotic producing bacteria from the soil samples collected from waste dumpsite. Among the 5 microbial isolates, 2 were shown to have inhibitory activities against Escherichia coli and Salmonella paratyphi. Morphological and biochemical tests revealed that both strains were Bacillus species with some differences in cultural characteristics. Molecular identification was performed by sequencing of the amplified 16S rRNA PCR products. The result showed that the two microbes were Bacillus subtilis and Bacillus cereus with 98% and 97% similarity score, respectively. This study suggests that Bacillus species have the potential to produce antibiotics against a broad spectrum of microbial growth and will be helpful in improving these strains for better production.
... For Bacillus and Staphylococcus APS-5 and APS-3, 6 showed zone of 14 mm and 10, 17 mm respectively (Fig. 5 and Table 5). In other study [13] investigated, two isolates 5 and 6 inhibited the growth of S. aureus and P. vulgris with zone having diameter of 11 and 12 mm respectively. ...
Antibiotics, potential against multi-drug resistant (MDR) pathogenic bacteria is currently become a vital research area. The present research approach is focused on the isolation of antibiotic producing bacteria from the soil samples collected from several diverse habitats viz. preserved areas of Himalayas, natural water resources, rhizosphere of medicinal plants, agricultural soil, forest soils, industrial waste soil from district Kullu and Poanta Sahib (HP). The bacterial isolates were characterized on the basis of their morphology, further confirmed by biochemical and haemolytic activity tests. Antibiotic producing ability of isolates was confirmed by zone of inhibition around bacterial colonies. A total of 30 isolates belonging to six different genera i.e. Micrococcus, Bacillus, Lactococcus, Pseudomonas, E.coli and Proteus, antibiotic producing bacteria were found in different soil samples. Out of these, 12 potential antibiotic producing strains showed their inhibitory response against pathogenic microorganisms. These results suggested that after proper standardization and processing, soil isolates having antibiotic producing capability can be used commercially.
... It exhibited remarkable antagonistic activity against different pathogenic test organisms. This species of Bacillus is an aerobic spore former found commonly in soil, groundwater, plants and animals during harvest or slaughter [22]. Studies based on the past decade analysis on antibiotic screening revealed most members of the genus Bacillus to have a contribution in the search for new antibiotics [23]. ...
Objective: The purpose of our study was to isolate and identify the bacteriocinogenic strain exhibiting broad range antimicrobial activity and to analyze the effect of different culturing conditions on the production of an antimicrobial metabolites isolated from the soil of Simlipal Biosphere Reserve, India. Methods: In the current study, bacterial strains were screened for antimicrobial activity from soil samples of five different regions. The effect of varying culture conditions such as pH, incubation period, and temperature along with carbon and nitrogen sources with and without certain salts was studied. The characterization of the potent strain was studied by morphological, biochemical, and 16S rRNA genetic sequencing. A phylogenetic affiliation of the strain was studied. Results: A total of 31 out of 245 strains isolated from soil were screened on the basis of antimicrobial results against the test pathogens. On the basis of bacteriocin-like inhibition studies method, one potential isolate that exhibited the highest inhibition against all the pathogens was selected. The optimization of highest antimicrobial metabolite production by the isolate with the influence of physical parameters was found as the incubation period of 3 days with 37°C temperature at pH 8 and for the chemical parameters dextrose was showed the most effective carbon sources when implemented with salts and yeast extract as the best sources of nitrogen with salts. The crude metabolite showed an absorbance peak value of 1.234 with optimum ƛ-max at 214 nm. The potent isolate showed maximum identity with Bacillus amyloliquefaciens (99% similarity) with highest query coverage on basic local alignment search tool search analysis of the 16S rDNA sequence. Phylogenetic analysis revealed close affiliation of the isolate with B. amyloliquefaciens (KC494392.1) having antimicrobial activity. Conclusion: The findings revealed that the incubation period, temperature, pH, and the culture medium have a direct influence on the production of metabolites. These parameters can be modified for the improvement of the fermentation process.
... Secondary metabolites specifically antibiotics produced by microorganisms have been very useful for the cure of certain human diseases caused by bacteria, fungi and protozoa. Most of antibiotic producers used today are soil microbes with fungal strains, streptomyces and bacteria being extensively used in industrial antibiotic production (Satvinder, 2014). ...
... Although these bacteria were not yet identified to the species level, morphological and biochemical characteristics indicate that they belong to the genus Burkholderia spp. ( [24]. ...
Objective: The objectives of this study were to isolate microorganisms and screen for potential antimicrobial activities from the soil. Methods: In this study, a total of 425 isolates were isolated from 100 soil samples. The preliminary screening for antimicrobial activities of these isolates was performed by modified cross-streak, agar diffusion, and modified icrodilution technique against 16 pathogenic bacteria and fungi.Results: In the anti-microbial activity, there were three isolates, namely, 277, 303, and 307 exhibited inhibitory activity against methicillin-resistantStaphylococcus aureus and Salmonella typhimurium respectively. This study also examined the various enzymes producing from soil microorganisms including chitinase, chitosanase, amylase, cellulose, caseinase, gelatinase, esterase, and lipase production of different selective media for 24 and 48hrs using the direct spot method. The results revealed that 28 isolates could produce various enzymes with strong activity. Most of them produced gelatinase (5.65%) and caseinase (5.18%). There were four isolates that produce broad-spectrum enzyme. In addition, the investigation of selectedmicroorganism identification showed that they can be divided into three groups: Burkholderia spp., Pseudomonas spp., and Rhodococcus spp.Conclusion: This study demonstrated that the microorganisms from soil are capable of producing potential, antibacterial, and bioactive enzymes.Keywords: Antimicrobial activity, Extracellular enzyme, Soil microbial, Drug-resistant bacteria.
... The first class of AMPs comprises ribosomally synthesized peptides, including bacteriocins whereas the second class comprises small microbial peptides synthesized enzymatically by non-ribosomal pathways (13). The Bacteriocins and Bacteriocin-like inhibitory substances (BLIS) contain lanthionine and/or methyllanthionine residues employed to form a ring through intramolecular post-translational modifications like subtilin, a 32-aminoacid pentacyclic lantibiotic (3320 Da) produced by B. subtilis ATCC 6633. ...
Background: Bacillus genus have been found to produce antimicrobial polypeptides that have a remarkable capability to combat the current increasing problem of antibiotic resistance. The aim of this cross sectional study was to determine the proportion and pattern of distribution of antibiotic producing and non-producing species of Bacillus in soil and also to analyse the activity of the antibiotic producers.
Methods: A total of 40 soil samples were collected from the wastelands of Kathmandu Valley and 121 isolates were studied from them. They were identified according to Bergey’s Manual. The antibiotic were extracted and the activity was analysed by agar- well diffusion method against the test organisms Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922 and Pseudomonas aeruginosa ATCC 27853.
Result: Bacillus subtilis species were found to be in highest number (17.3%) and Bacillus macerans (0.82%) in lowest number. 18.33% of Bacillus isolates were found to produce antibiotics. The highest number of antibiotic producing isolate was B. subtilis (40.09%) and the least was B. circulans (4.54%). The number of antibiotic producing bacteria were not found to be affected by moisture content of soil and pH change. Among the antibiotic producers, 63.63% produced antibiotic against Gram positive bacteria, 27.27% against Gram negative bacteria and B. alvei (9.09%) had a broad spectrum activity. Some colonies were even found to produce antibiotic with a greater potential than the conventional antibiotics.
Conclusion: Based on property of soil, moist-neutral soil had the highest number of bacilli isolates and dry-acidic and moist-alkaline had least number of isolates.
DOI: 10.21276/AABS.2017.1316
... Our result was correlated with Kaur et al. who isolated antibiotics producing microorganisms from soil in Punjab, India. These microorganisms were investigating based on the disc diffusion method, agar well method, streak agar method and found that there were 2 isolates suitable in inhibition the growth of Staphylococcus aureus and Proteus vulgris indicator strains [30]. Inhibition zone from modified cross-streak method in the table is derived from 3 experiments and given in mean±SD (n=3). ...
Objective: Our study was to isolate Bacillus spp. from soil all around 6 geographic parts of Thailand and screen for potential antimicrobial.Methods: A total of 43 isolates which isolated from 100 samples of soil were investigated. Preliminary screening was based on antimicrobial activity against 16 strains of pathogenic bacteria and fungi, including 10 strains of Methicillin-resistant Staphylococcus aureus (MRSA), and 1 strain of Methicillin-sensitivity Staphylococcus aureus (MSSA), Extended spectrum beta lactamases (ESBL) Escherichia coli, Samonella Typhymurium, Klebsiella pneumoniae, Candida albicans, Cryptococcus neoformans respectively by cross streak, agar diffusion and modified microdilution technique. Moreover, the selected Bacillus spp. were then screened for bioactive enzyme, including chitinese, chitosanese, amylase, cellulose, caseinase, gelatinase, esterase and lipase production of different selective media for 24 and 48 h by direct spot agar.Results: There are 2 isolates, namely 23 and 49 showed particularly strong activity inhibitions against MRSA and pathogenic C. albicans. The diameter of hydrolysis zone results from screened for bioactive enzyme revealed that there were 36 isolate showed particularly strong activities with a broad spectrum enzyme. The isolate which produced the widest diameter hydrolysis zone in gelatin, cellulose and starch are 294, 303, and 290 respectively.Conclusion: Our result indicated Bacillus spp. from soil in Thailand showed potent antibacterial activity and bioactive compounds production.
One of the most essential and widely used secondary metabolites produced by bacteria, fungus, and Streptomyces is the antibiotic. The soil is the best place to find microorganisms that produce antibiotics. Soil is a diverse supply of bacteria that produce antibiotics because it is complex, heterogeneous and home to a variety of microorganisms. The majority of antibiotics used today are produced in artificial media, isolated from soil microorganisms and extracted. With the use of an antibiotic produced by a bacterium, some human diseases caused by bacteria, fungus, and protozoa can be healed. The antifungal and antibacterial capabilities of fungi like Penicillium and Cephalosporium make them important sources of secondary metabolites and vital for the synthesis of industrial enzymes. Because they are more affordable and effective at stopping the growth of other organisms, the common soil bacteria known as bacillus species have been proven to be beneficial. The population of microorganisms is dependent on a number of characteristics, including the type of soil, temperature, water-holding capacity, oxygen pressure, pH, carbon supply, salt concentration, organic matter, aeration, moisture content, and others.
To investigate the influence of microbial wax removal and viscosity reduction technologies on the fluidity of waxy crude oil, this study took the waxy crude oil from China's Daqing Oilfield as the experimental object for microbial wax removal and viscosity reduction experiment. First, utilizing standard strain screening and culture protocols, a high-efficiency microbial strain H-1 (Ochrobactrum intermedium bacterium) was proposed from in situ soil. The single factor experiment and multi-factor response surface analysis were then utilized to optimize the temperature, initial pH value of the medium, and NaCl mass concentration needed for the strain's growth. Oil displaced activity and emulsification tests were used to evaluate the biosurfactants produced by the metabolism of strain H-1, and Fourier transform infrared spectroscopy was used to identify the types of biosurfactants. Finally, the application effect of strain H-1 on wax removal and viscosity reduction of waxy crude oil were clarified through the wax crystal morphology, wax concentration, and viscosity of waxy crude oil. The research findings indicated that strain H-1 produced best at a temperature of 38 °C, pH value was 7.1 and the mass fraction of NaCl was 0.65%. The lipopeptide biosurfactant produced by strain H-1 can release oil. Additionally, strain H-1 has a high emulsifying capacity, with a 61.25% emulsifying coefficient. After 7 days of treatment with the strain H-1, the wax crystal morphology of the waxy crude oil changed from a large-sized aggregated to a small-sized sparse condition, and the wax content of the waxy crude oil fell by 38.67%. The viscosity reduction rate of strain H-1 to waxy crude oil was consistently greater than 30% over the temperature range of 30–42 °C, with the greatest rate of 42.38% at 39 °C. The preceding results demonstrate that strain H-1 can degrade the wax components of waxy crude oil, lower the crude oil's low temperature viscosity, and then improve the crude oil's low-temperature fluidity.
Objective: To isolate and screen antibiotic producing actinomycetes from potential soil samples
of Gondar town, Ethiopia.
Methods: Fifteen soil samples were collected, serially diluted and spread on starch casein
and oat meal agar supplemented with amoxicillin and cyclohexamide for inhibition of bacteria
and fungi, respectively. Cross streak method was used to check antagonistic activity of isolated
actinomycetes against test organisms. Solid state fermentation and crude extraction were used for
the production of antibiotics from isolates. Agar well diffusion was used for antimicrobial activity
of crude extracts against test organisms.
Results: Three isolates (Ab18, Ab28 and Ab43) have been shown high antagonistic activity during
primary screening. Inhibition zones obtained from crude extracts showed significance differences
when compared with standard antibiotics tested against test organisms (P
One hundred seventy-seven actinomycetes strains were isolated from soils collected from fruit orchards in Thailand. All were tested for antibacterial activity against seven pathogenic bacteria using co-cultivation methods. Forty strains (22.6%) were active against at least one indicator bacteria. Twenty-seven strains (15.3%) inhibited only gram-positive bacteria, four strains (2.3%) inhibited only gram-negative bacteria, and nine strains (5.1%) showed activity against both. Strain SJE177 had potent activity against all tested bacteria, and was selected for further investigation. A crude ethyl acetate extract of this strain retained inhibitory activity as tested by disk-diffusion method. Analysis of morphological and biochemical characteristics and the 16S rRNA gene sequence indicated this strain belonged to the genus Streptomyces. The strain formed a monophyletic line in a phylogenetic tree of 16S rRNA gene sequences with other Streptomyces reference strains. High performance liquid chromatography (HPLC) analysis showed SJE177 produced actinomycin. Since many isolates showed inhibitory activity against indicator bacteria, these results suggest Thai soil could be an interesting source to explore for antibacterial substances.
To help select the most appropriate method for detecting indole production with anaerobic gram-negative bacilli, several recommended methods were compared. Indole was measured both quantitatively and qualitatively after varying periods of incubation. Studies evaluated the results obtained in different media, the effect of adding glucose and/or tryptophan, the requirement for strict anaerobiasis, and the effects of reducing the total volume of broth. A 1-ml amount of thioglycolate broth without glucose but with 0.02% tryptophan gave optimal results after 2 to 7 days of incubation in anaerobic (Gas-Pak) jars. The majority of clinical isolates will give strong positive tests after 1 to 2 days but a few require 3 to 7 days of incubation. Prolonged incubation was required more frequently with conventional methods.
Naturally soil is rich in microorganisms capable of antibiotic synthesis but the frequency with which synthesis occurs at ecologically significant levels in has been much less clear. Over the past decade, however, genetic and molecular techniques, have been applied to demonstrate conclusively that microorganisms synthesize a variety of antibiotics, even under field conditions, in the rhizosphere. Soil sample from the Post Graduate Hostel of the Permanent Site campus, University of Ilorin, Nigeria was screened for antibioticproducing microorganisms by agar sensitivity assay. Seven bacterial species and one fungus were isolated. The bacterial species were identified by their cellular characteristics, colonial morphology and biochemical tests. The bacterial isolates include; Staphylococcus aureus, Proteus vulgaris, Bacillus spp., Pseudomonas aeruginosa, Micrococcus luteus, Escherichia coli and Micrococcus varians. The fungus was identified to be Rhizopus stolonifer using lactophenol - in- cotton blue staining technique and possessed inhibitory action against test isolates. Only Bacillus spp exhibited antibacterial activity of all the bacteria isolated.
Antibioticsβ-Lactam antibioticsTetracycline groupRifamycinsAminoglycoside-aminocyclitol antibioticsMacrolidesLincosamidesStreptograminsPolypeptide antibioticsGlycopeptide antibioticsMiscellaneous antibacterial antibioticsAntifungal antibioticsSynthetic antimicrobial agentsAntiviral drugsDrug combinations
A simple and rapid semiquantitative slide catalase test useful for the identification of oxidative and nonsaccharolytic gram-negative bacteria, i.e., "nonfermenters," is described. Using the interpretative criterion of time of appearance of oxygen bubbles in 3% hydrogen peroxide, three categories of nonfermenters were established. The rapid catalase producers included Achromobacter xylosoxidans and Achromobacter species; Acinetobacter anitratus and Acinetobacter lwoffii; Bordetella bronchiseptica; CDC group IVE; Pseudomonas aeruginosa, P. fluorescens, P. putida, P. diminuta, and P. acidovorans; and Moraxella urethralis and M-6. The delayed catalase producers included Bordetella parapertussis, CDC group VA-1, P. alcaligenes, P. cepacia, P. mendocina, P. pickettii (VA-2), P. pseudoalcaligenes, P. putrefaciens, P. stutzeri, P. testosteroni, and P. vesicularis. The third group consisted of an additional 17 taxa of nonfermenters which were classified as moderate catalase producers.
To help select the most appropriate method for detecting indole production with anaerobic gram-negative bacilli, several recommended methods were compared. Indole was measured both quantitatively and qualitatively after varying periods of incubation. Studies evaluated the results obtained in different media, the effect of adding glucose and/or tryptophan, the requirement for strict anaerobiasis, and the effects of reducing the total volume of broth. A 1-ml amount of thioglycolate broth without glucose but with 0.02% tryptophan gave optimal results after 2 to 7 days of incubation in anaerobic (Gas-Pak) jars. The majority of clinical isolates will give strong positive tests after 1 to 2 days but a few require 3 to 7 days of incubation. Prolonged incubation was required more frequently with conventional methods.
The actinomycetes are well known as a group of filamentous, Gram-positive bacteria that produce many useful secondary metabolites, including antibiotics and enzymes. Although they have been intensively studied for both theoretical and practical objectives, there is much scope for developing our basic knowledge of the means of detection and isolation of these microbes. This session concentrated on new methods for the detection and identification of novel actinomycetes from a range of environments. Approaches to the detection of actinomycetes ranged from investigations of neglected habitats and extreme environments (e.g. alkaline soils and oil drills) to the analysis of DNA extracted from the environment and use of specific phages. The continuing problems of the identification of actinomycete isolates were also considered. Topics discussed included use of phage typing, DNA probes, and correlation between phenetic and genotypic species of Streptomyces.
Biochemical Tests for Identification of Medical Bacteria
- J F Macfaddin
MacFaddin, JF. (1980). Biochemical Tests for
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