scholarly journals Sodium Azide Induced Mutation in Actinomycetes

2018 ◽  
Vol 4 ◽  
pp. 39-42
Author(s):  
Muna Tamang ◽  
Pujan K.C. ◽  
Punya Kumari Koju ◽  
Puspa Lachhimasyu ◽  
Dinesh Dhakal ◽  
...  
Keyword(s):  
Sodium Azide ◽  
Test Organism ◽  
Soil Samples ◽  
Loss Of Function ◽  
Wild Type ◽  
Sugar Utilization ◽  
E Coli ◽  
Streptomyces Spp ◽  
Microbiological Methods ◽  
Gain And Loss

Objectives: The study was done with an aim to determine the gain and loss of functions among the actinomycetes mutants induced by sodium azide.Methods: The study was carried out in the laboratory of the Sainik Awasiya Mahavidhayala, Bhaktapur, Nepal from 2016 December to 2017 March. A total of 30 soil samples were collected from Tokha, Bhaktapur area and Godawari area. Actinomycetes were isolated from the soil sample using pour plate technique on selective media; starch casein agar. The isolates were identified by using standard microbiological methods and each isolate was exposed to different concentration of sodium azide to generate mutants. The wild type and mutants were compared in morphology, biochemical reactions and antibiotic susceptibility to test organism to determine the gain and loss of functions.Results: Among 30 samples processed, 20(67%) actinomycetes were isolated, in which 6 (20%) were identified as the Streptomyces spp. A total of 28 mutants were isolated from 6 wild types by exposed at 10ppm, 20ppm, 40ppm, 50ppm, 100ppm concentration of sodium azide. Out of 28 mutants formed, only 10 mutants from sample showed same pigmentation as its wild type while other 18 mutants showed change in their pigmentation. In sugar utilization test, 8 different sugars for 28 mutants each, 56 cases showed Gain of Function (GOF), similarly 44 cases showed Loss of function (LOF). Antibiosis remained unaffected against Pseudomonas i.e. no GOF or LOF was seen. Only 2 cases of LOF against Staphylococcus aureus were seen while there were no cases of LOF in other pathogens. 3 cases of GOF against E. coli, 4 against S Typhi and 4 against S. aureus were observed.Conclusion: The potential of mutant actinomycetes has been realized, and hence opens exciting avenues in the field of biotechnology and biomedical research.

scholarly journals Fermentation and Extraction of Antibacterial Metabolite Using Streptomyces spp. Isolated from Taplejung, Nepal

2021 ◽  
Vol 26 (1) ◽  
pp. 8-15
Author(s):  
Shiv Nandan Sah ◽  
Ramesh Majhi ◽  
Sunil Regmi ◽  
Arjun Ghimire ◽  
Bhageshwor Biswas ◽  
...  
Keyword(s):  
Antibacterial Activity ◽  
Salmonella Typhi ◽  
Soil Samples ◽  
E Coli ◽  
Primary Screening ◽  
Streptomyces Spp ◽  
Antibacterial Compound ◽  
Fermented Broth ◽  
Layer Chromatography ◽  
Mic Value

Realizing an increasing need for a novel antibiotic, this study was carried out to screen antibacterial metabolites producing actinomycetes from 15 soil samples collected from Taplejung. Antibacterial metabolites producing actinomycetes were confirmed by primary screening and secondary screening. Macroscopic, microscopic, and biochemical characteristics were used for presumptive identification of probable actinomycetes genera. The potential isolate was cultured in starch casein broth for production of possible antibacterial compound. The antibacterial compound was extracted from fermented broth using organic solvents like ethyl acetate, n-butanol, chloroform, dichloromethane, and methanol. Among 24 isolates, only one (T18) showed antibacterial activity against both Gram-positive (Staphylococcus aureus and Bacillus subtilis) and Gram-negative (Escherichia coli, Salmonella Typhi and Pseudomonas aeruginosa) test-bacteria. The isolate was considered as Streptomyces spp based on microscopy and various biochemical, and physiological characteristics. Extracted antibacterial metabolite showed antibacterial activity with a MIC value of 1.2 mg/mL against E. coli (ATCC 25922). The chromatogram in Thin Layer Chromatography showed only one spot exhibited by extract with Rf value 0.87 suggested that the isolate produced a compound that was completely different from the spot with Rf value 0.94 produced by gentamicin (standard). This study revealed the distribution of the potent antibacterial metabolite producing actinomycetes in the soils of Taplejung.

scholarly journals ANTIBACTERIAL ACTIVITY FOR BIOSURFACTANT PRODUCED BY STREPTOMYCES SPP. ISOLATED FROM SOIL SAMPLES

2020 ◽  
pp. 109-110
Author(s):  
SAMER M. AL-HULU
Keyword(s):  
Antibacterial Activity ◽  
Aerial Mycelium ◽  
Inhibition Zone ◽  
Activity Analysis ◽  
Soil Samples ◽  
Blood Agar ◽  
Malt Extract ◽  
E Coli ◽  
Streptomyces Spp ◽  
High Degree

Objective: The goal of the present study on isolation Streptomyces from soil samples with biosurfactant activity and antibacterial activity analysis. Methods: (25) The soil samples were taken form Hilla city. (10) It is Streptomyces spp. They were identified. (4) Streptomyces spp. having the ability to hemolysis on blood agar that has the capacity to generate biosurfactants. Streptomyces spp. 8 displayed a high degree of activity by having a blood agar inhibition zone (25 mm). Results: Antibacterial activity was evaluated for Streptomyces spp. 8 (Staphylococcus aureus, Escherichia Coli, and Pseudomonas aeruginosa). Streptomyces spp. 8 has greatest inhibition zone against S. aureus equal to 14 mm compared 12 mm against E. coli and 8 mm for P. aeruginosa. This Streptomyces spp. 8 characteristic was tested, it showed Gram positive with aerial mycelium gray in color on yeast malt extract agar. Negative for melanin produced on tyrosine broth medium, negative for H2S output, and pigment development, it has for the use of mannitol sucrose, glucose, and fructose as carbon source and negative for xylose. Conclusion: Isolated Streptomyces spp. having the potential to generate biosurfactants with antibacterial activity.

scholarly journals Investigation of the in vitro and in vivo activity of the type IV pilus extension ATPase BfpD

2020 ◽  
Author(s):  
Jinlei Zhao ◽  
Shahista Nisa ◽  
Michael S. Donnenberg
Keyword(s):  
Atpase Activity ◽  
Loss Of Function ◽  
Wild Type ◽  
Multifunctional Protein ◽  
Mutant Proteins ◽  
E Coli ◽  
Type Iv ◽  
Kinetics Of

AbstractType IV pili (T4Ps) are multifunctional protein fibers found in many bacteria and archaea. All T4P systems have an extension ATPase, which provides the energy required to push structural subunits out of the membrane. We previously reported that the BfpD T4P ATPase from enteropathogenic E. coli (EPEC) has the expected hexameric structure and ATPase activity, the latter enhanced by the presence of the N-terminal cytoplasmic domains of its partner proteins BfpC and BfpE. In this study, we further investigated the kinetics of the BfpD ATPase. Despite high purity of the proteins, the reported enhanced ATPase activity was found to be from (an) ATPase(s) contaminating the N-BfpC preparation. Furthermore, although two mutations in highly conserved bfpD sites led to loss of function in vivo, the purified mutant proteins retained some ATPase activity, albeit less than the wild-type protein. Therefore, the observed ATPase activity of BfpD was also affected by (a) contaminating ATPase(s). Expression of the mutant bfpD alleles did not interfere with BfpD function in bacteria that also expressed wild-type BfpD. However, a similar mutation of bfpF, which encodes the retraction ATPase, blocked the function of wild-type BfpF when both were present. These results highlight similarities and differences in function and activity of T4P extension and retraction ATPases in EPEC.

scholarly journals Natural IgA and TNFRSF13B polymorphism: a double edged sword fueling balancing selection

2021 ◽  
Author(s):  
Jeffrey L. Platt ◽  
Mayara Garcia de Mattos Barbosa ◽  
Daniel Huynh ◽  
Adam R. Lefferts ◽  
Juhi Katta ◽  
...  
Keyword(s):  
High Frequency ◽  
Virulence Genes ◽  
Balancing Selection ◽  
Dominant Negative ◽  
Loss Of Function ◽  
Wild Type ◽  
E Coli ◽  
High Prevalence ◽  
Protective Antibodies ◽  
Dominant Negative Variant

AbstractTNFRSF13B encodes the “transmembrane-activator and CAML-interactor” (TACI) receptor, which drives plasma cell differentiation. Although TNFRSF13B supports host defense, dominant-negative TNFRSF13B alleles are common in humans and other species and only rarely associate with disease. We reasoned the high frequency of disruptive TNFRSF13B alleles reflects balancing selection, the loss of function conferring advantage in some settings. Testing that concept, we asked whether and how a common human dominant negative variant, TNFRSF13B A181E, imparts resistance to enteric pathogens. Mice engineered to express mono-allelic or bi-allelic A144E variants of tnrsf13B, corresponding to A181E exhibited striking resistance to pathogenicity and transmission of C. rodentium, a murine pathogen that models enterohemorrhagic E. coli, and resistance was principally owed to deficiency of natural IgA in the intestine. In wild type mice with gut IgA and in mutant mice fed IgA, binding of Ig induces expression of LEE encoded virulence genes, which confer pathogenicity and transmission. C. rodentium and probably some other enteric organisms thus appropriated binding of otherwise protective antibodies to signal induction of the virulence program and the high prevalence of TNFRSF13B dominant negative variants thus reflects balancing selection.

Transport of escherichia coli through soil to groundwater traced by randomly amplified polymorphic DNA (RAPD)

1997 ◽  
Vol 35 (11-12) ◽  
pp. 351-357 ◽  
Author(s):  
R. Rothmaier ◽  
A. Weidenmann ◽  
K. Botzenhart
Keyword(s):  
Negative Impact ◽  
Random Amplified Polymorphic Dna ◽  
Coli Strain ◽  
Soil Samples ◽  
Test Field ◽  
Liquid Manure ◽  
E Coli ◽  
Rapd Pattern ◽  
Geological Situation ◽  
Different Strains

Isolates (50) of E. coli obtained from liquid manure (20 bovine, 20 porcine) were genotyped using random amplified polymorphic DNA (RAPD). Typing revealed 9 and 14 different strains in bovine and porcine liquid manure respectively with no strains in common. One porcine strain, showing a simple RAPD pattern, was subcultured and spread on a test field (1.5l/m2 at 1010 cfu/l) in a drinking water protection zone with loamy to sandy sediments in the Donauried area, Baden-Wurttemberg. Soil samples and groundwaters were collected at monthly intervals October 1994 – June 1995 during which 114 E. coli isolates were recovered. The first occurrence and maximum concentration of E. coli in soil samples taken from more than 20cm depth was in January 1995, declining rapidly with depth and time. All isolates from soil and only one from groundwater showed the RAPD pattern of the spread E. coli strain. The results could not demonstrate a severe negative impact of the spreading of liquid manure on the bacteriological quality of the groundwater in the given geological situation. The distinct strain patterns found in different kinds of liquid manure suggest that genotyping of E. coli by RAPD may be an adequate tool for tracing sources of faecal contamination.

The Effect of Mianserin on Lifespan of Caenorhabditis elegans is Abolished by Glucose

2021 ◽  
Vol 13 ◽  
Author(s):  
Abdullah Almotayri ◽  
Jency Thomas ◽  
Mihiri Munasinghe ◽  
Markandeya Jois
Keyword(s):  
Caenorhabditis Elegans ◽  
Scaffolding Protein ◽  
Lifespan Extension ◽  
Wild Type ◽  
E Coli ◽  
C Elegans ◽  
Risk Of Death ◽  
Increased Risk ◽  
Antidepressant Use ◽  
Extension Effect

Background: The antidepressant mianserin has been shown to extend the lifespan of Caenorhabditis elegans (C. elegans), a well-established model organism used in aging research. The extension of lifespan in C. elegans was shown to be dependent on increased expression of the scaffolding protein (ANK3/unc-44). In contrast, antidepressant use in humans is associated with an increased risk of death. The C. elegans in the laboratory are fed Escherichia coli (E. coli), a diet high in protein and low in carbohydrate, whereas a typical human diet is high in carbohydrates. We hypothesized that dietary carbohydrates might mitigate the lifespan-extension effect of mianserin. Objective: To investigate the effect of glucose added to the diet of C. elegans on the lifespan-extension effect of mianserin. Methods: Wild-type Bristol N2 and ANK3/unc-44 inactivating mutants were cultured on agar plates containing nematode growth medium and fed E. coli. Treatment groups included (C) control, (M50) 50 μM mianserin, (G) 73 mM glucose, and (M50G) 50 μM mianserin and 73 mM glucose. Lifespan was determined by monitoring the worms until they died. Statistical analysis was performed using the Kaplan-Meier version of the log-rank test. Results: Mianserin treatment resulted in a 12% increase in lifespan (P<0.05) of wild-type Bristol N2 worms but reduced lifespan by 6% in ANK3/unc-44 mutants, consistent with previous research. The addition of glucose to the diet reduced the lifespan of both strains of worms and abolished the lifespan-extension by mianserin. Conclusion: The addition of glucose to the diet of C. elegans abolishes the lifespan-extension effects of mianserin.

scholarly journals Mutations in fbiD (Rv2983) as a Novel Determinant of Resistance to Pretomanid and Delamanid in Mycobacterium tuberculosis

2020 ◽  
Vol 65 (1) ◽  
pp. e01948-20
Author(s):  
Dalin Rifat ◽  
Si-Yang Li ◽  
Thomas Ioerger ◽  
Keshav Shah ◽  
Jean-Philippe Lanoix ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Clinical Evidence ◽  
Clinical Samples ◽  
Loss Of Function ◽  
Wild Type ◽  
Content Type ◽  
Solid Media ◽  
High Level ◽  
Level Resistance

ABSTRACTThe nitroimidazole prodrugs delamanid and pretomanid comprise one of only two new antimicrobial classes approved to treat tuberculosis (TB) in 50 years. Prior in vitro studies suggest a relatively low barrier to nitroimidazole resistance in Mycobacterium tuberculosis, but clinical evidence is limited to date. We selected pretomanid-resistant M. tuberculosis mutants in two mouse models of TB using a range of pretomanid doses. The frequency of spontaneous resistance was approximately 10−5 CFU. Whole-genome sequencing of 161 resistant isolates from 47 mice revealed 99 unique mutations, of which 91% occurred in 1 of 5 genes previously associated with nitroimidazole activation and resistance, namely, fbiC (56%), fbiA (15%), ddn (12%), fgd (4%), and fbiB (4%). Nearly all mutations were unique to a single mouse and not previously identified. The remaining 9% of resistant mutants harbored mutations in Rv2983 (fbiD), a gene not previously associated with nitroimidazole resistance but recently shown to be a guanylyltransferase necessary for cofactor F420 synthesis. Most mutants exhibited high-level resistance to pretomanid and delamanid, although Rv2983 and fbiB mutants exhibited high-level pretomanid resistance but relatively small changes in delamanid susceptibility. Complementing an Rv2983 mutant with wild-type Rv2983 restored susceptibility to pretomanid and delamanid. By quantifying intracellular F420 and its precursor Fo in overexpressing and loss-of-function mutants, we provide further evidence that Rv2983 is necessary for F420 biosynthesis. Finally, Rv2983 mutants and other F420H2-deficient mutants displayed hypersusceptibility to some antibiotics and to concentrations of malachite green found in solid media used to isolate and propagate mycobacteria from clinical samples.

scholarly journals The NALCN Channel Regulator UNC-80 Functions in a Subset of Interneurons To Regulate Caenorhabditis elegans Reversal Behavior

2019 ◽  
Vol 10 (1) ◽  
pp. 199-210 ◽  
Author(s):  
Chuanman Zhou ◽  
Jintao Luo ◽  
Xiaohui He ◽  
Qian Zhou ◽  
Yunxia He ◽  
...  
Keyword(s):  
Plant Hormone ◽  
Neuronal Excitability ◽  
Resting Membrane Potential ◽  
Loss Of Function ◽  
Wild Type ◽  
C Elegans ◽  
Link Type ◽  
Complex Functions ◽  
And Function ◽  
Multiple Loss

NALCN (Na+leak channel, non-selective) is a conserved, voltage-insensitive cation channel that regulates resting membrane potential and neuronal excitability. UNC79 and UNC80 are key regulators of the channel function. However, the behavioral effects of the channel complex are not entirely clear and the neurons in which the channel functions remain to be identified. In a forward genetic screen for C. elegans mutants with defective avoidance response to the plant hormone methyl salicylate (MeSa), we isolated multiple loss-of-function mutations in unc-80 and unc-79. C. elegans NALCN mutants exhibited similarly defective MeSa avoidance. Interestingly, NALCN, unc-80 and unc-79 mutants all showed wild type-like responses to other attractive or repelling odorants, suggesting that NALCN does not broadly affect odor detection or related forward and reversal behaviors. To understand in which neurons the channel functions, we determined the identities of a subset of unc-80-expressing neurons. We found that unc-79 and unc-80 are expressed and function in overlapping neurons, which verified previous assumptions. Neuron-specific transgene rescue and knockdown experiments suggest that the command interneurons AVA and AVE and the anterior guidepost neuron AVG can play a sufficient role in mediating unc-80 regulation of the MeSa avoidance. Though primarily based on genetic analyses, our results further imply that MeSa might activate NALCN by direct or indirect actions. Altogether, we provide an initial look into the key neurons in which the NALCN channel complex functions and identify a novel function of the channel in regulating C. elegans reversal behavior through command interneurons.

New insights in gut-liver axis in wild-type murine imiquimod-induced lupus

Lupus ◽  
2021 ◽  
Vol 30 (6) ◽  
pp. 926-936
Author(s):  
Georges Maalouly ◽  
Joelle Hajal ◽  
Charbel Noujeim ◽  
Michel Choueiry ◽  
Hussein Nassereddine ◽  
...  
Keyword(s):  
Tight Junction ◽  
Fecal Calprotectin ◽  
Liver Inflammation ◽  
Tight Junction Proteins ◽  
Wild Type ◽  
Imiquimod Cream ◽  
E Coli ◽  
Intestinal Pathology ◽  
Nfκb Pathway ◽  
Junction Proteins

Background Intestinal and hepatic manifestations of lupus seem to be underestimated in comparison to other major organ lesions. Although recent data point to gut-liver axis involvement in lupus, gut permeability dysfunction and liver inflammation need to be more investigated. Objective This study aims to assess fecal calprotectin, intestinal tight junction proteins and liver inflammation pathway in wild-type murine imiquimod- induced lupus. Methods C57BL/6 mice were topically treated on their right ears with 1.25 mg of 5% imiquimod cream, three times per week for six weeks. Fecal calprotectin was collected at day 0, 22 and 45. Renal, liver and intestinal pathology, as well as inflammatory markers, intestinal tight junction proteins, and E. coli protein in liver were assessed at sacrifice. Results At six weeks, lupus nephritis was confirmed on histopathology and NGAL and KIM-1 expression. Calprotectin rise started at day 22 and persists at day 45. Protein expression of Claudine, ZO-1 and occludin was significantly decreased. E. coli protein was significantly increased in liver with necro-inflammation and increased TLR4, TLR7, and pNFκB/NFκB liver expression. Conclusion This study is the first to demonstrate early fecal calprotectin increase and liver activation of TLR4- NFκB pathway in wild-type murine imiquimod-induced lupus.

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