scholarly journals Ozonetherapy as a new antimicrobic strategy

2020 ◽  
pp. 195-200
Author(s):  
R. Shahanenko ◽  
N. Ilnitskiy ◽  
V. Shahanenko ◽  
S. Rublenko
Keyword(s):  
Ozone Concentration ◽  
Antimicrobial Properties ◽  
Nacl Solution ◽  
Ozone Therapy ◽  
E Coli ◽  
Purulent Wounds ◽  
Microbial Number

Development of antibiotic-resistant strains of microorganisms is a dangerous phenomenon, actively progressing every year. The uncontrolled use of antibiotics for animals, accumulation in products of animal origin ultimately poses a danger to human health. That is why a decrease in the use of antibiotics and searching alternatives of antibiotic is acute and relevant issues. Therefore, the aim of our research was to study the antimicrobial properties of ozone in relation to pathogens of purulent infection and to show the possibility of using ozone therapy as a potential method of antimicrobial therapy for animals. The materials for determining antimicrobial effect of ozone was 12 samples of purulent exudate in an amount of 2 ml, taken from dogs with purulent wounds before and after sanitation by ozonized 0.87% NaCl solution. Complexity course of wound process with purulent inflammation largely depends on from degree of microbial contamination of the wound and species composition of microorganisms. Therefore, an important aspect in our research was the study of the antimicrobial properties of ozone on its action of purulent exudate «in vitro» and «in vivo» and the determination of the bactericidal effect on microorganisms. The most stable and informative indicator of assessing nature of purulent-inflammatory process is the determination of total number of microorganisms in 1 ml of discharge from a purulent wound. The total microbial number was determined by the method of serial dilutions according to Pasteur. Serial ten-fold dilutions from 10-1 to 10-9 were prepared from purulent exudate in test tubes with sterile MPB (9 ml each). Species composition of microorganisms was determined by cultural and biochemical properties of cultivated microbial colonies, followed microscopy of smears from pure cultures stained using method of Gram. Samples of purulent exudate were subjected to microbiological examination before treatment, and after 30 minutes of washing by ozonized isotonic solution NaCl (ozone concentration of 7 mg/ml). A microbiological study of purulent exudate was also carried out, pre-treated with ozone at a concentration of 7 mg/ml «in vitro» in a test tube in an amount of 2 ml by passing it through exudate (sparging) at a flow rate of 0.5 L/min and a processing time of 10 min. Samples were examined immediately after sampling and sparging. "Microbial landscapes" of purulent wounds were presented by associations of Staph. aureus, Str. faecalis, E. coli. Microbial seeding of purulent exudate for treatment ranged from 6.6 • 10-10 to 3.7 • 10-8 CFU/ml, however, after 10 min of bubbling «in vitro» at an ozone concentration of 7 mg/ml, the degree of microbial seeding of samples did not exceed 10-4 CFU/ml and ranged from 3.1 • 10-4 to 2.3• 10-3 CFU/ml. As shown by the results of microbiological studies, the growth of microorganisms on a nutrient medium in bacteriological plates with purulent exudate samples treated with ozone with concentration of 7 mg/ml was already absent at 10-5 degrees of dilution, which indicates the pronounced antimicrobial properties of ozone. The study «in vivo»also indicates that even after a single use of an ozonized isotonic NaCl solution at an ozone concentration of 7 mg/ml, it completely prevents the growth of Staph. aureus, Str. faecalis, E. coli. and causes 100% death mentioned associations of microorganisms. The results of microbiological studies are confirmed by clinical data. So, on the third day of treatment, the animals in the lesion zone had a small amount of wound exudate, and the microbial number of the latter was 1.4 • 10-4 − 3.1 • 10-³ CFU/ml, below the critical level of contamination and in most cases not leads to the progression of a purulent-inflammatory process. Ozone destroys all types of bacteria, viruses, fungi and protozoa. At the same time, ozone at a concentration of 7 mg/ ml does not have an irritating effect on body tissues, therefore, ozone therapy can be considered as an additional or alternative therapy of bacterial infection. Key words: ozone, ozonetherapy, purulent wounds, antibiotic resistance.

scholarly journals Antimicrobial, antioxidant activities and toxicity on Cavia porcellus of Dialium angolense Welw. Ex Oliv, a traditional medicinal plant from Bagira in Eastern of DR Congo

2020 ◽  
Vol 13 (2) ◽  
pp. 166-180
Author(s):  
Bashige Chiribagula V ◽  
Bakari Amuri S ◽  
Okusa Ndjolo Philippe ◽  
Kahumba Byanga J ◽  
Duez P ◽  
...  
Keyword(s):  
Antioxidant Activity ◽  
Acute Toxicity ◽  
Antioxidant Activities ◽  
Antimicrobial Properties ◽  
Salmonella Typhi ◽  
Antimicrobial Activities ◽  
Cavia Porcellus ◽  
E Coli

Dialium angolense is used in Bagira for its various medicinal properties particularly in the management of infectious diseases. In this study, the methanol and aqueous extracts of leaves and fruits were evaluated for their in vitro antioxidant and antimicrobial properties and their in vivo toxicity on Cavia porcellus. The major phytochemical classes of extracts were screened using standard in-tube reactions. The antimicrobial study was tested on Candida albicans, Escherichia coli, Salmonella typhi, Staphylococcus aureus and Streptococcus pneumoniae using agar well diffusion and dilution methods, while the antioxidant activity was evaluated by a DPPH assay. For the acute toxicity study, animals (6/group) were orally given in a single dose 5000, 1000 or 15000 mg of extract/kg body weight (BW) then observed for 14 days. In sub-acute toxicity assays, 150 or 300 mg/kg BW/day were orally given, and animals observed for 28 days. Total phenolics and total flavonoids contents ranged 1.19 to 1.61 mg GAE.g-1 and 0.45 to 1.01 mg QEg-1, respectively. The extracts presented antioxidant activity with IC50 ranging 4.9 to 6.9 µg/mL. The minimal inhibitory concentration (MIC) on tested strains ranged from 1.9 to 500 µg/mL with the aqueous extract of fruits as a most active extract: MIC=1.9 µg/mL on E. coli and C. albicans. No signs of toxicity were noted during the acute and sub-acute toxicity assessments, suggesting a maximal tolerate doses (MDT) and LD50 > 15000 mg/kg BW. This study highlights the antioxidant and antimicrobial activities of Dialium angolense and suggests that further studies be directed towards the isolation of active compounds.

scholarly journals Anthocyanin as potential source for antimicrobial activity inClitoria ternateaL. andDioscorea alataL.

2018 ◽  
Vol 47 (6) ◽  
pp. 490-495 ◽  
Author(s):  
Noraini Mahmad ◽  
R.M. Taha ◽  
Rashidi Othman ◽  
Sakinah Abdullah ◽  
Nordiyanah Anuar ◽  
...  
Keyword(s):  
Antimicrobial Properties ◽  
Blue Flower ◽  
Distilled Water ◽  
Content Type ◽  
E Coli ◽  
Inhibition Zones ◽  
Antibacterial And Antifungal ◽  
Purple Yam

PurposeThe purpose of this paper is to validate the antimicrobial activity (both antibacterial and antifungal) ofin vivoandin vitroethanolic anthocyanin extracts ofClitoria ternateaL. (vivid blue flower butterfly-pea) andDioscorea alataL. (purple yam) against selected bacteria (Bacillus subtilis,Staphylococcus aureusandEscherichia coli) and fungi (Fusarium sp.,Aspergillus nigerandTrichoderma sp.).Design/methodology/approachThe freeze-dried samples (0.2 g) fromin vivovivid blue flowers ofC. ternateaL. were extracted using 10 mL ethanol (produced ethanolic red extraction) and 10 mL distilled water (produced aqueous blue extraction) separately. Two-month-oldin vitrocallus samples (0.2 g) were only extracted using 10 mL ethanol. The anthocyanin extractions were separated with the addition (several times) of ethyl acetate and distilled water (1:2:3) to remove stilbenoids, chlorophyll, less polar flavonoids and other non-polar compounds. Furthermore, the antimicrobial properties were determined using agar diffusion technique. Three bacteria (B. subtilis,S. aureusandE. coli) and fungi (F. sp.,A. nigerandT. sp.) were streaked on bacteria agar and dextrose agar, respectively, using “hockey stick”. Then, the sterile paper discs (6 mm diameter) were pipetted with 20 µL of 1,010 CFU/mL chloramphenicol (as control for antibacterial) and carbendazim (as control for antifungal)in vivoandin vitroextracts. The plates were incubated at room temperature for 48 h, and the inhibition zones were measured.FindingsBased on the results, bothin vivoandin vitroethanolic extracts from vivid blue flowers ofC. ternateaL. showed the best antibacterial activity against the same bacteria (B. subtilis), 11 and 10 mm inhibition zones, respectively. However, different antifungal activity was detected inin vitroethanolic callus extract (12 mm), which was againstT. sp., contrary toin vivoethanolic extract (10 mm), which was againstF. sp.; antibacterial activity ofD. alataL. was seen against the same bacteria (E. coli) with the highest inhibition zone forin vivoextract (8.8 mm), followed byin vitroextract (7.8 mm).Research limitations/implicationsAnthocyanins are responsible for the water soluble and vacuolar, pink, red, purple and blue pigments present in coloured plant pigments. These pigments (pink, red, purple and blue) are of important agronomic value in many crops and ornamental plants. However, anthocyanins are not stable and are easy to degrade and fade whenever exposed to light.Social implicationsPlant extracts containing bioactive agents with antimicrobial properties have been found to be useful in treating bacterial and fungal infections, as well as showed multiple antibiotic resistance.Originality/valueBothin vivoandin vitroextracts from vivid blue flower petals (C. ternateaL.) and purple yam (D. alataL.) have important applications as natural antimicrobial (antibacterial and antifungal) agents in the coating industry, instead of natural pharmaceutical products.

scholarly journals Identification of two domains and distal histidine ligands to the four haems in the bacterial c-type cytochrome NapC; the prototype connector between quinol/quinone and periplasmic oxido-reductases

2002 ◽  
Vol 368 (2) ◽  
pp. 425-432 ◽  
Author(s):  
Michaël L. CARTRON ◽  
M. Dolores ROLDÁN ◽  
Stuart J. FERGUSON ◽  
Ben C. BERKS ◽  
David J. RICHARDSON
Keyword(s):  
Electron Transfer ◽  
Paracoccus Denitrificans ◽  
Spectroscopic Studies ◽  
Water Soluble ◽  
Broad Absorption Band ◽  
Binding Motifs ◽  
E Coli

NapC is a tetra-haem member of a family of bacterial membrane-anchored multi-haem c-type cytochromes implicated in electron transfer between membrane quinols and periplasmic enzymes. The water-soluble tetra-haem fragment of Paracoccus pantotrophus NapC has been expressed as a periplasmic protein (NapCsol) in Paracoccus denitrificans, P. pantotrophus and Escherichia coli. Site-specific mutagenesis of NapCsol, combined with spectroscopic studies, suggests that each haem iron centre has bis-histidinyl co-ordination. Four proximal ligands arise from each of four Cys-Xaa-Xaa-Cys-His haem-binding motifs; candidates for the four distal ligands are His81, His99, His174 and His194. NapCH81A, NapCH99A, NapCH174A and NapCH194A mutants (with alanine substituted for each of the four candidate residues) have all been purified from E. coli. In each case, one of the haems has become high-spin, as judged by the presence of a broad absorption band between 620nm and 650nm for the oxidized cytochrome; this feature is absent for wild-type protein and presumably arises because of the absence of the distal histidine ligand from one of the haems. NapCH81A and NapCH174A are less well expressed in E. coli than NapCH99A and NapCH194A and cannot be detected when expressed in P. denitrificans or P. pantotrophus. In vitro and in vivo complementation studies demonstrate that the soluble periplasmic NapC can mediate electron transfer from quinols to the periplasmic nitrate reductase. This capacity was retained in vitro with the NapCH99A and NapCH194A mutants but was lost in vivo. A model for the structural organization of NapCsol into two domains, each containing a di-haem pair, is proposed. In this model, each haem pair obtains one distal haem ligand from its own domain and a second from the other domain. The suggestion of two domains is supported by observations that the 24kDa NapCsol cleaves to yield a 12kDa haem-staining band. Determination of the cleavage site showed it was between two equally sized di-haem domains predicted from sequence analysis.

scholarly journals Isolation and Characterization of a New T-Even Bacteriophage, CEV1, and Determination of Its Potential To Reduce Escherichia coli O157:H7 Levels in Sheep

2006 ◽  
Vol 72 (9) ◽  
pp. 6405-6410 ◽  
Author(s):  
Raul R. Raya ◽  
Peter Varey ◽  
Rebecca A. Oot ◽  
Michael R. Dyen ◽  
Todd R. Callaway ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Oral Dose ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Isolation And Characterization ◽  
Unit Reduction

ABSTRACT Bacteriophage CEV1 was isolated from sheep resistant to Escherichia coli O157:H7 colonization. In vitro, CEV1 efficiently infected E. coli O157:H7 grown both aerobically and anaerobically. In vivo, sheep receiving a single oral dose of CEV1 showed a 2-log-unit reduction in intestinal E. coli O157:H7 levels within 2 days compared to levels in the controls.

A Molecular Approach to Immunoscintigraphy: A Study of the T-Antigen Conformation on the Surface of Tumors

1987 ◽  
Vol 26 (01) ◽  
pp. 1-6 ◽  
Author(s):  
S. Selvaraj ◽  
M. R. Suresh ◽  
G. McLean ◽  
D. Willans ◽  
C. Turner ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Tumor Cell ◽  
T Antigen ◽  
Human Carcinomas ◽  
Animal Tumor ◽  
Synthetic Antigens

The role of glycoconjugates in tumor cell differentiation has been well documented. We have examined the expression of the two anomers of the Thomsen-Friedenreich antigen on the surface of human, canine and murine tumor cell membranes both in vitro and in vivo. This has been accomplished through the synthesis of the disaccharide terminal residues in both a and ß configuration. Both entities were used to generate murine monoclonal antibodies which recognized the carbohydrate determinants. The determination of fine specificities of these antibodies was effected by means of cellular uptake, immunohistopathology and immunoscintigraphy. Examination of pathological specimens of human and canine tumor tissue indicated that the expressed antigen was in the β configuration. More than 89% of all human carcinomas tested expressed the antigen in the above anomeric form. The combination of synthetic antigens and monoclonal antibodies raised specifically against them provide us with invaluable tools for the study of tumor marker expression in humans and their respective animal tumor models.

Storage of Human Blood Platelets

1974 ◽  
Vol 32 (02/03) ◽  
pp. 405-416 ◽  
Author(s):  
M. R Hardeman ◽  
Carina J L. Heynens
Keyword(s):  
Storage Temperature ◽  
Platelet Rich Plasma ◽  
Blood Platelets ◽  
Storage Period ◽  
Serotonin Uptake ◽  
Hypotonic Shock ◽  
Glycolytic Intermediates

SummaryStorage experiments were performed at 4°, 25° and 37° C with platelet-rich plasma under sterile conditions. In some experiments also the effect of storing platelets at 4° C in whole blood was investigated.Before, during and after three days of storage, the platelets were tested at 37° C for their serotonin uptake and response to hypotonic shock. In addition some glycolytic intermediates were determined.A fair correlation was noticed between the serotonin uptake and hypotonic shock experiments. Both parameters were best maintained at 25° C. Also platelet counting, performed after the storage period, indicated 25° C as the best storage temperature. Determination of glycolytic intermediates did not justify any conclusion regarding the optimal storage temperature. Of the various anticoagulants studied, ACD and heparin gave the best results as to the serotonin uptake and hypotonic shock response, either with fresh or stored platelets. The use of EDTA resulted in the lowest activity, especially after storage.The results of these storage experiments in vitro, correspond well with those in vivo reported in the literature.

Rapid, Refined, and Robust Method for Expression, Purification, and Characterization of Recombinant Human Amyloid-beta M1-42

2019 ◽  
Author(s):  
Priya Prakash ◽  
Travis Lantz ◽  
Krupal P. Jethava ◽  
Gaurav Chopra
Keyword(s):  
Amyloid Beta ◽  
Western Blots ◽  
Force Microscopy ◽  
E Coli ◽  
Atomic Force ◽  
Set Up ◽  
Short Time

Amyloid plaques found in the brains of Alzheimer’s disease (AD) patients primarily consists of amyloid beta 1-42 (Ab42). Commercially, Ab42 is synthetized using peptide synthesizers. We describe a robust methodology for expression of recombinant human Ab(M1-42) in Rosetta(DE3)pLysS and BL21(DE3)pLysS competent E. coli with refined and rapid analytical purification techniques. The peptide is isolated and purified from the transformed cells using an optimized set-up for reverse-phase HPLC protocol, using commonly available C18 columns, yielding high amounts of peptide (~15-20 mg per 1 L culture) in a short time. The recombinant Ab(M1-42) forms characteristic aggregates similar to synthetic Ab42 aggregates as verified by western blots and atomic force microscopy to warrant future biological use. Our rapid, refined, and robust technique to purify human Ab(M1-42) can be used to synthesize chemical probes for several downstream in vitro and in vivo assays to facilitate AD research.

scholarly journals Functional identification of ygiP as a positive regulator of the ttdA-ttdB-ygjE operon

Microbiology ◽  
2006 ◽  
Vol 152 (7) ◽  
pp. 2129-2135 ◽  
Author(s):  
Taku Oshima ◽  
Francis Biville
Keyword(s):  
Functional Characterization ◽  
Anaerobic Growth ◽  
Functional Identification ◽  
New Members ◽  
E Coli ◽  
Inner Membrane Protein ◽  
Tartrate Dehydratase

Functional characterization of unknown genes is currently a major task in biology. The search for gene function involves a combination of various in silico, in vitro and in vivo approaches. Available knowledge from the study of more than 21 LysR-type regulators in Escherichia coli has facilitated the classification of new members of the family. From sequence similarities and its location on the E. coli chromosome, it is suggested that ygiP encodes a lysR regulator controlling the expression of a neighbouring operon; this operon encodes the two subunits of tartrate dehydratase (TtdA, TtdB) and YgiE, an integral inner-membrane protein possibly involved in tartrate uptake. Expression of tartrate dehydratase, which converts tartrate to oxaloacetate, is required for anaerobic growth on glycerol as carbon source in the presence of tartrate. Here, it has been demonstrated that disruption of ygiP, ttdA or ygjE abolishes tartrate-dependent anaerobic growth on glycerol. It has also been shown that tartrate-dependent induction of the ttdA-ttdB-ygjE operon requires a functional YgiP.

Carbon dots derived from Fusobacterium nucleatum for intracellular determination of Fe3+ and bioimaging both in vitro and in vivo

2021 ◽  
Author(s):  
Lijuan Liu ◽  
Shengting Zhang ◽  
Xiaodan Zheng ◽  
Hongmei Li ◽  
Qi Chen ◽  
...  
Keyword(s):  
Carbon Dots ◽  
Fusobacterium Nucleatum ◽  
Living Cells ◽  
First Time ◽  
Fluorescent Carbon Dots ◽  
Fluorescent Carbon

Fusobacterium nucleatum has been employed for the first time to synthesize fluorescent carbon dots which could be applied for the determination of Fe3+ ions in living cells and bioimaging in vitro and in vivo with excellent biocompatibility.

scholarly journals Arabidopsis Disulfide Reductase, Trx-h2, Functions as an RNA Chaperone under Cold Stress

2021 ◽  
Vol 11 (15) ◽  
pp. 6865
Author(s):  
Eun Seon Lee ◽  
Joung Hun Park ◽  
Seong Dong Wi ◽  
Ho Byoung Chae ◽  
Seol Ki Paeng ◽  
...  
Keyword(s):  
Cold Stress ◽  
Wild Type ◽  
Rna Chaperone ◽  
E Coli ◽  
Cold Sensitive ◽  
Thioredoxin H ◽  
Functional Rnas

The thioredoxin-h (Trx-h) family of Arabidopsis thaliana comprises cytosolic disulfide reductases. However, the physiological function of Trx-h2, which contains an additional 19 amino acids at its N-terminus, remains unclear. In this study, we investigated the molecular function of Trx-h2 both in vitro and in vivo and found that Arabidopsis Trx-h2 overexpression (Trx-h2OE) lines showed significantly longer roots than wild-type plants under cold stress. Therefore, we further investigated the role of Trx-h2 under cold stress. Our results revealed that Trx-h2 functions as an RNA chaperone by melting misfolded and non-functional RNAs, and by facilitating their correct folding into active forms with native conformation. We showed that Trx-h2 binds to and efficiently melts nucleic acids (ssDNA, dsDNA, and RNA), and facilitates the export of mRNAs from the nucleus to the cytoplasm under cold stress. Moreover, overexpression of Trx-h2 increased the survival rate of the cold-sensitive E. coli BX04 cells under low temperature. Thus, our data show that Trx-h2 performs function as an RNA chaperone under cold stress, thus increasing plant cold tolerance.

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