scholarly journals Isolation and Molecular Characterization of Antibiotic Producing Bacillus licheniformis Strains Isolated from Soil

2020 ◽  
Vol 14 (4) ◽  
pp. 2363-2370
Author(s):  
Anhar Al-Turk ◽  
Nidal Odat ◽  
Muhannad I. Massadeh

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.

2020 ◽  
pp. 24-30

Calotropis procera is shrub of Asclepiadaceae family and is a source of antimicrobial metabolites against a greater range of bacteria including Streptococcus Group B (SGB). In the present study, different metabolic compounds were extracted from C. procera using 75% methanol. The extract of C. procera was assessed against different bacteria of the SGB family by estimating the zone of inhibition. Moreover, minimal inhibitory concentration (MIC) of different concentrations e.g., 0.001, 0.01, 0.1, 1.0 and 10 mg/mL were determined through zone of inhibition. Gas chromatography mass spectrometry (GC-MS) analysis of plant extract revealed 24 compounds. The most important compounds of the GC-MS array were including undecane, terephthalic acid, Cyclohexane, dimethyl-propane-thiosulfinate, Fluorobenzoic acid Octadecenoic acid and others. The antibacterial activities of the plant extracts were might be because of their compound which had been reported previously as well as an antimicrobial compound. The molecular characterization of different bacteria of SGB used in the study revealed the bacteria were consisted of antibiotic resistant genes against.


Author(s):  
Anthony Kayode Onifade ◽  
Sunday Stephen Abi

Two hundred and fifty-six (256) human faecal samples were collected from thirty-three (33) underground faecal storage cisterns across 5 major towns (Akure, Akungba, Ondo, Ore and Owo) in Ondo State, Nigeria. Salmonella-Shigella agar, Eosine Methylene Blue agar and MacConkey agar, which are selective for faecal bacteria, were used to isolate a total of 103 strains of Gram-negative bacteria. Biochemical characterization of the isolates revealed Escherichia coli (32.04%), Enterobacter aerogenes (14.56), Klepsiella pneumoniae (12.62%), Salmonella typhi (11.65%), Psudomonas aeruginosa (7.77%), Shigella dysenteriae (6.80%), Proteus mirabilis (5.83%), Citrobacter koseri (3.88%), Providentia alcalifaciens (2.91%) and Klepsiella oxytoca (1.94%).  All the bacterial isolates were then subjected to antibiotic sensitivity test using antibiotic discs impregnated with Augmentin (30 µg), Gentamycin (10 µg), Pefloxacin (10 µg), Ofloxacin (5 µg), Streptomycin (10 μg), Sulfomethoxazole-trimethoprim (30 µg), Chloramphenicol (30 μg); Sparfloxacin (10 μg); Ciprofloxacin (5 μg) and Amoxicillin (20 μg). The resistance of the isolates to the antibiotics used varied from 27.18% to Ciprofloxacin, 33.98% to Ofloxacin, 47.57% to Sulphomethazole-Trimethroprim, 56.31% to Streptomycin, 66.99% to Chloramphenicol, 74.76% to Pefloxacin, 75.73% to Sparfloxacin, 75.73% to Gentamycin, 85.44% to Ampicillin and 95.15 to Augmentin. These results showed that underground human faecal storage cisterns contain potential multiple antibiotic resistant bacteria that may be transmitted to human via formites, animal vectors and water.


Antibiotics ◽  
2019 ◽  
Vol 8 (3) ◽  
pp. 136 ◽  
Author(s):  
Ndegwa ◽  
Almehmadi ◽  
Kim ◽  
Kaseloo ◽  
Ako

There is a scarcity of information on antibiotic resistance in goats. To understand shedding of resistant Escherichia coli in pastured goats, we collected fecal samples from a mixed age cohort over a one-year period. No antibiotic had been used on the study animals one year prior to and during the study period. Resistant isolates were detected in all age groups and prevalence in goat kids was significantly higher than adults; 43–48% vs 8–25% respectively. The proportion of resistant isolates was higher when animals were congregated near handling facility than on pasture. Most isolates were resistant to tetracycline (51%) and streptomycin (30%), but also to antibiotics that had never been used on the farm; ampicillin (19%). TetB, bla-TEM, (aadA and strpA/strpB) genes were detected in 70%, 43%, (44% and 24%) of tetracycline, ampicillin, and streptomycin resistant isolates respectively. Resistant isolates also harbored virulent genes and some belonged to D and B2 phylogenetic groups. Thus, pastured goats, despite minimal exposure to antibiotics, are reservoirs of resistant E. coli that may contaminate the environment and food chain and spread resistant genes to pathogenic bacteria and some that are potential animal and human pathogens. Environmental sources may play a role in acquisition of resistant bacteria in pastured goats.


2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Oluwakayode O. Odeja ◽  
Michael Gabriel Ibok ◽  
Ejike O. Okpala

Abstract Background Asparagus flagellaris leaves are ethnomedicinally used to treat syphilis, gonorrhea and other sexually transmitted diseases (STDs), with no reports on the volatile constituents. This study was aimed to quantitatively and qualitatively characterise the composition of essential oil, evaluates the free radical scavenging and antimicrobial capacity of the essential oil. Methods The essential oil was isolated by hydrodistillation method using all-glass Clevenger-type apparatus, while the identification and quantification of constituents were performed by gas chromatography-mass spectrometry (GC-MS) technique. The antioxidant activity on 2, 2-diphenyl-1-picrylhydrazyl (DPPH) was evaluated and the oil was also tested against 10 strains of microorganisms consisting of 6 bacteria: Escherichia coli, Salmonella typhi, Klebsiella pneumonia, Pseudomonas aeruginosa, Staphylococcus aureus and Bacillus subtilis and 4 fungi: Candida albicans, Penicillium notatum, Aspergillus niger and Rhizopus spp. using broth dilution and surface plate methods, respectively. Results A pale yellow essential oil with a characteristic scent was obtained, with a yield of 0.80% (w/w). A total of 28 compounds accounting for 97.41% of the total oil contents were identified. The oil was predominated by Thymol and its derivatives, accounting for 57.48%. The most abundance (% area) constituents of the essential oil were 5-Thymyl tiglate (18.49%), Thymyl-2-methyl butyrate (17.34%), Thymol hydroquinone dimethyl ether (10.52%), Thymol methyl ether (9.42%) and 5-Propyl-1, 3-benzodioxole (4.59%). The essential oil showed a significant free radical scavenging activity compared to the standard antioxidant drugs used in this study, with % inhibition varying from 88.06 ± 0.0001 to 93.05 ± 0.0006. The leaf essential oil exhibited antimicrobial activity on all the tested organisms at 500–125 μg/mL, with an 18–10 mm inhibitory zone. Conclusion The leaf essential oil of A. flagellaris contains notable chemical compounds responsible for its antioxidant and antimicrobial activities.


Author(s):  
Meesha Singh ◽  
Rupsha Karmakar ◽  
Sayak Ganguli ◽  
Mahashweta Mitra Ghosh

Aims: This study aims at comparative identification of antibiotic resistance patterns in bacteria isolated from samples collected from rural environment (LS) and urban environments (SS). Metagenomic profiling gave us insights into the microbial abundance of the two samples. This study focused on culture-based methods for complete identification of antibiotic resistant isolates and estimation of comparative antibiotic resistance among the two samples. Study Design: Untreated medical waste and anthropogenic waste disposal can lead to the propagation of different antibiotic resistant strains in wastewater environments both in urban and rural set ups which provide an insight towards this study approach mentioned in the methodology segment. Place and Duration of Study: Sewer system of a medical facility located in Purulia, India was the collection site for liquid sludge. Solid sludge and associated wastewater were collected in vicinity of a large urban medical facility from central Kolkata, India. Methodology: Physico-chemical properties were analyzed followed by microbiological and biochemical characterization. The antibiotic resistance patterns were determined by Kirby-Bauer disc diffusion assay. Potent multidrug resistant isolates were identified using 16srRNA gene amplification followed by Phylogenetic profiling, using CLC Genomics workbench. Results: We observed maximum resistance in an E. coli isolate which was resistant up to 22 antibiotics. Combined data for resistance from urban and rural samples were found to exhibit 83.9% resistance to beta lactams, 85.7% to macrolides, 44.2% to fluoroquinolones, 50% to glycopeptides and cephalosporins, 35.7 % to carbapenems and sulfonamides, 28.5 % to tetracycline, and 23.8 % to aminoglycosides. Conclusion: The high prevalence of antibiotic-resistant bacteria harbouring diverse resistance traits across samples indicated towards probable horizontal gene transfer across environmental niches. This study can prove to be useful to understand and map the patterns of resistance and stringently apply the counter measures related to public health practices.


Antibiotics ◽  
2021 ◽  
Vol 10 (9) ◽  
pp. 1044
Author(s):  
Bahman Khameneh ◽  
N. A. Michael Eskin ◽  
Milad Iranshahy ◽  
Bibi Sedigheh Fazly Bazzaz

The extensive usage of antibiotics and the rapid emergence of antimicrobial-resistant microbes (AMR) are becoming important global public health issues. Many solutions to these problems have been proposed, including developing alternative compounds with antimicrobial activities, managing existing antimicrobials, and rapidly detecting AMR pathogens. Among all of them, employing alternative compounds such as phytochemicals alone or in combination with other antibacterial agents appears to be both an effective and safe strategy for battling against these pathogens. The present review summarizes the scientific evidence on the biochemical, pharmacological, and clinical aspects of phytochemicals used to treat microbial pathogenesis. A wide range of commercial products are currently available on the market. Their well-documented clinical efficacy suggests that phytomedicines are valuable sources of new types of antimicrobial agents for future use. Innovative approaches and methodologies for identifying plant-derived products effective against AMR are also proposed in this review.


2020 ◽  
Vol 75 (9) ◽  
pp. 2554-2563 ◽  
Author(s):  
Christopher Fröhlich ◽  
Vidar Sørum ◽  
Sandra Huber ◽  
Ørjan Samuelsen ◽  
Fanny Berglund ◽  
...  

Abstract Background MBLs form a large and heterogeneous group of bacterial enzymes conferring resistance to β-lactam antibiotics, including carbapenems. A large environmental reservoir of MBLs has been identified, which can act as a source for transfer into human pathogens. Therefore, structural investigation of environmental and clinically rare MBLs can give new insights into structure–activity relationships to explore the role of catalytic and second shell residues, which are under selective pressure. Objectives To investigate the structure and activity of the environmental subclass B1 MBLs MYO-1, SHD-1 and ECV-1. Methods The respective genes of these MBLs were cloned into vectors and expressed in Escherichia coli. Purified enzymes were characterized with respect to their catalytic efficiency (kcat/Km). The enzymatic activities and MICs were determined for a panel of different β-lactams, including penicillins, cephalosporins and carbapenems. Thermostability was measured and structures were solved using X-ray crystallography (MYO-1 and ECV-1) or generated by homology modelling (SHD-1). Results Expression of the environmental MBLs in E. coli resulted in the characteristic MBL profile, not affecting aztreonam susceptibility and decreasing susceptibility to carbapenems, cephalosporins and penicillins. The purified enzymes showed variable catalytic activity in the order of <5% to ∼70% compared with the clinically widespread NDM-1. The thermostability of ECV-1 and SHD-1 was up to 8°C higher than that of MYO-1 and NDM-1. Using solved structures and molecular modelling, we identified differences in their second shell composition, possibly responsible for their relatively low hydrolytic activity. Conclusions These results show the importance of environmental species acting as reservoirs for MBL-encoding genes.


Viruses ◽  
2020 ◽  
Vol 12 (2) ◽  
pp. 133 ◽  
Author(s):  
Joseph M. Ochieng’ Oduor ◽  
Ermir Kadija ◽  
Atunga Nyachieo ◽  
Marianne W. Mureithi ◽  
Mikael Skurnik

Emergence of antibiotic-resistant bacteria is a serious threat to the public health. This is also true for Staphylococcus aureus and other staphylococci. Staphylococcus phages Stab20, Stab21, Stab22, and Stab23, were isolated in Albania. Based on genomic and phylogenetic analysis, they were classified to genus Kayvirus of the subfamily Twortvirinae. In this work, we describe the in-depth characterization of the phages that electron microscopy confirmed to be myoviruses. These phages showed tolerance to pH range of 5.4 to 9.4, to maximum UV radiation energy of 25 µJ/cm2, to temperatures up to 45 °C, and to ethanol concentrations up to 25%, and complete resistance to chloroform. The adsorption rate constants of the phages ranged between 1.0 × 10−9 mL/min and 4.7 × 10−9 mL/min, and the burst size was from 42 to 130 plaque-forming units. The phages Stab20, 21, 22, and 23, originally isolated using Staphylococcus xylosus as a host, demonstrated varied host ranges among different Staphylococcus strains suggesting that they could be included in cocktail formulations for therapeutic or bio-control purpose. Phage particle proteomes, consisting on average of ca 60–70 gene products, revealed, in addition to straight-forward structural proteins, also the presence of enzymes such DNA polymerase, helicases, recombinases, exonucleases, and RNA ligase polymer. They are likely to be injected into the bacteria along with the genomic DNA to take over the host metabolism as soon as possible after infection.


2020 ◽  
Vol 8 (1) ◽  
pp. 138 ◽  
Author(s):  
Pierre Philippe Mbehang Nguema ◽  
Richard Onanga ◽  
Guy Roger Ndong Atome ◽  
Jean Constant Obague Mbeang ◽  
Arsène Mabika Mabika ◽  
...  

In Gabon, terrestrial mammals of protected areas have been identified as a possible source of antibiotic-resistant bacteria. Some studies on antibiotic resistance in bats have already been carried out. The main goal of our study was to detect extended-spectrum beta-lactamases (ESBLs) that are produced by enterobacteria from bats in the Makokou region in Gabon. Sixty-eight fecal samples were obtained from 68 bats caught in the forests located 1 km from the little town of Makokou. After culture and isolation, 66 Gram-negative bacterial colonies were obtained. The double-disk diffusion test confirmed the presence of ESBLs in six (20.69%) Escherichia coli isolates, four (13.79%) Klebsiella pneumoniae isolates, and one (3.45%) Enterobacter cloacae isolate. The analysis based on the nucleotide sequences of the ESBL resistance genes showed that all cefotaximase-Munichs (CTX-Ms) were CTX-M-15 and that all sulfhydryl variables (SHVs) were SHV-11: 41.67% CTX-M-15-producing E. coli, 16.67% CTX-M-15+SHV-11-producing E. coli, 8.33% CTX-M-15-producing K. pneumoniae, 25% CTX-M-15+SHV-11-producing K. pneumoniae, and 8.33% CTX-M-15-produced E. cloacae. This study shows for the first time the presence of multiresistant ESBL-producing enterobacteria in fruit bats in Makokou.


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