scholarly journals Isolation of Helicobacter pylori from Gastric Biopsy of Dyspeptic Patients in Ghana and In Vitro Preliminary Assessment of the Effect of Dissotis rotundifolia Extract on Its Growth

2018 ◽  
Vol 2018 ◽  
pp. 1-6 ◽  
Author(s):  
Michael Buenor Adinortey ◽  
Charles Ansah ◽  
Cynthia Ayefoumi Adinortey ◽  
Ansumana Sandy Bockarie ◽  
Martin Tangnaa Morna ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Therapeutic Potential ◽  
Gastric Biopsy ◽  
Zone Of Inhibition ◽  
Triphenyltetrazolium Chloride ◽  
Gram Negative ◽  
Successful Isolation ◽  
Pure Cultures ◽  
H Pylori

Helicobacter pylori (H. pylori) is a gram-negative bacterium that colonizes the human stomach. Infection with this microaerophilic bacterium causes gastric and duodenal ulcer. This study sought to isolate H. pylori, from gastric biopsy samples of dyspeptic patients in Ghana using a 2,3,5-triphenyltetrazolium chloride (TTC) dye incorporated medium method. This TTC dye method was further used in an antimicrobial susceptibility assay involving Dissotis rotundifolia extract (DRE). H. pylori were successfully isolated from gastric biopsy of dyspeptic patients. Pure cultures of H. pylori in 2,3,5-triphenyltetrazolium chloride (TTC) dye incorporated medium were seen as sparkling colonies. Isolates, identified as H. pylori, were gram-negative and urease, catalase, and oxidase positive and showed characteristic morphology as spiral-shaped bacteria under the microscope. The organisms were found to be susceptible to cephalothin and resistant to nalidixic acid. Above all, the observation that H. pylori grew only at 37°C and not 25°C or 42°C affirms that the bacterium is neither Helicobacter cinaedi nor Helicobacter fenneliae. The anti-H. pylori study depicts a statistically lower zone of inhibition for DRE compared to standard drugs [amoxicillin and clarithromycin] (p<0.05), whereas metronidazole showed no zone of inhibition. This study reports the first successful isolation and culturing of H. pylori in Ghana using TTC dye. It also shows that DRE possess an in vitro anti-H. pylori activity and that DRE has some therapeutic potential against H. pylori infection.

scholarly journals In Vitro Anti-Helicobacter pyloriActivities of New Rifamycin Derivatives, KRM-1648 and KRM-1657

1999 ◽  
Vol 43 (5) ◽  
pp. 1072-1076 ◽  
Author(s):  
Junko K. Akada ◽  
Mutsunori Shirai ◽  
Kenji Fujii ◽  
Kiwamu Okita ◽  
Teruko Nakazawa
Keyword(s):  
Helicobacter Pylori ◽  
Cell Lysis ◽  
Antimicrobial Activities ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Bactericidal Activities ◽  
H Pylori ◽  
Time Kill

ABSTRACT The new rifamycin derivatives KRM-1657 and KRM-1648 were evaluated for their in vitro antimicrobial activities against 44 strains ofHelicobacter pylori. Although the drugs were not very active against other gram-negative bacteria, the MICs at which 90% of isolates are inhibited for these drugs were lower (0.002 and 0.008 μg/ml, respectively) than those of amoxicillin and rifampin forH. pylori. Time-kill studies revealed that the bactericidal activities of these agents were due to cell lysis. The results presented here indicate that these new rifamycin derivatives may be useful for the eradication of H. pylori infections.

scholarly journals Successful isolation of Helicobacter pylori after prolonged incubation: A case report of prolonged incubation for H. pylori

2012 ◽  
Vol 64 (4) ◽  
pp. 1297-1299
Author(s):  
Tatjana Babic ◽  
Biljana Miljkovic-Selimovic ◽  
Branislava Kocic ◽  
A. Nagorni ◽  
Ljiljana Ristic
Keyword(s):  
Helicobacter Pylori ◽  
Normal Growth ◽  
Gastric Biopsy ◽  
Gram Stain ◽  
Ulcus Ventriculi ◽  
Prolonged Incubation ◽  
Primary Isolation ◽  
Successful Isolation ◽  
Gastric Biopsies ◽  
H Pylori

The culture of Helicobacter pylori from a gastric biopsy is the ?gold standard? in the diagnosis of H. pylori infection. However, the primary isolation of H. pylori from gastric biopsies is rather demanding. The duration of incubation for the isolation of H. pylori has been recommended to be five to seven days: in the present case, we found that a prolonged incubation period allowed the successful isolation of H. pylori from a patient with ulcus ventriculi. Biopsies were placed directly into transport medium and processed for culture within two hours. On day 14, one suspected H. pylori-like colony appeared on one of the plates. The isolate was confirmed to be H. pylori based on its typical colony morphology, negative Gram stain, and positive urease, catalase and oxidase tests. The isolate, requiring 14 days recovery, later exhibited the normal growth characteristics of H. pylori strains, indicating its unusually long incubation requirement was a temporary predicament.

scholarly journals Bactericidal effect of Neem (Azadirachta indica) leaf extract on Helicobacter pylori

2021 ◽  
Vol 42 (6) ◽  
pp. 1591-1597
Author(s):  
A. Saxena ◽  
◽  
P. Arivaradarajan ◽  
A.K. Mukhopadhyay ◽  
S.P. Nandi ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Ethyl Acetate ◽  
Azadirachta Indica ◽  
Ethanolic Extract ◽  
Ethanol Extract ◽  
Zone Of Inhibition ◽  
Gram Negative ◽  
Neem Leaves ◽  
Gastric Pathogen ◽  
H Pylori

Aim: The aim of the present study was to investigate the antibacterial effect of ethanolic extract of neem (Azadirachta indica) leaf against Gram-negative, gastric pathogen, Helicobacter pylori (H. pylori). Methodology: Extracts of neem leaf were prepared in different solventslike hexane, dichloromethane, chloroform, ethyl acetate, acetone and ethanol. Antibacterial activity was estimated in terms of zone of inhibition by performing Agar cup diffusion assay. Depending on the diameter of zone of inhibition, ethyl acetate, acetone and ethanol extract of neem leaves were selected for Thin Layer Chromatography. The presence of photochemicals were detected using iodine fumigation. Elution Assay was done to detect the bioactive components of the ethanol extract. Results: Out of sixsolvents used, ethanol extract of neem leavesshowed the maximum zone of inhibition against H. pylori. TLC separation of ethyl acetate, acetone and ethanol extract of plant products showed dark brown bands of phytochemicals on silica-gel G 60 plates. The contact bioautography assay showed a zone of 15 mm. Elution assay and agar cup bioassay was performed against H. pylori and the loading spot showed a zone of 11 mm. Interpretation: The findings of the present study revealed the anti-bacterial potency of ethanolic extracts of neem (Azadirachta indica) leaf against Gram-negative gastric pathogen H. pylori. The ethanolic extract of neem leaf can be used as an effective natural remedy in combating H. pylori infection.

scholarly journals Engineered Endolysin-based "Artilysins" for Controlling the Gram-negative Pathogen Helicobacter Pylori

2021 ◽  
Author(s):  
Dengyuan Xu ◽  
Shanshan Zhao ◽  
Jun Dou ◽  
Xiaofeng Xu ◽  
Yanyan Zhi ◽  
...  
Keyword(s):  
Cell Wall ◽  
Helicobacter Pylori ◽  
Transmembrane Protein ◽  
Gastrointestinal Diseases ◽  
Amide Bond ◽  
Resistant Bacteria ◽  
Gram Negative ◽  
Antibiotic Overuse ◽  
H Pylori

Abstract Helicobacter pylori infection can cause a variety of gastrointestinal diseases. In severe cases, there is a risk of gastric cancer. Antibiotics are often used for clinical treatment of H. pylori infections. However, because of antibiotic overuse in recent years and the emergence of multidrug-resistant bacteria, there is an urgent need to develop new treatment methods and drugs to achieve complete eradication of H. pylori. Endolysins and holins encoded by bacterial viruses (i.e., phages) represent a promising avenue of investigation. These lyase-based antibacterial drugs act on the bacterial cell wall to destroy the bacteria. Currently, a type of endolysin that has been studied more frequently acts on the amide bond between peptidoglycans, and holin is a transmembrane protein that can punch holes in the cell membrane. However, as a Gram-negative bacterium, H. pylori possesses a layer of impermeable lipopolysaccharides on the cell wall, which prevents endolysin interaction with the cell wall. Therefore, we designed a genetic linkage between an endolysin enzyme and a holin enzyme with a section of polypeptides (e.g., polycations and hydrophobic peptides) that enable penetration of the outer membrane. These complexes were designated “artilysins” and were efficiently expressed in Escherichia coli. In vitro bacteriostasis experiments showed that the purified artilysins had strong bacteriostatic effects on H. pylori. In addition, the surface of H. pylori was perforated and destroyed, as confirmed by electron microscopy, which was proved that artilysins had bacteriolytic effect on H. pylori.

scholarly journals ANTIMICROBIAL ACTIVITY OF ANISOCHILUS CARNOSUS (L.F.) WALL AGAINST THE HUMAN GASTRIC PATHOGEN HELICOBACTER PYLORI

2017 ◽  
Vol 10 (10) ◽  
pp. 292
Author(s):  
Vignesh Shetty ◽  
Richard Lobo ◽  
Nimmy Kumar ◽  
Ramachandra Lingadakai ◽  
Ganesh C Pai ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Skin Diseases ◽  
Therapeutic Potential ◽  
High Elevation ◽  
Gastrointestinal Infections ◽  
Treatment Regimens ◽  
Soxhlet Apparatus ◽  
Soxhlet Method ◽  
H Pylori

  Objective: The prevalence of Helicobacter pylori in India is high, and majority leads to severe gastrointestinal infections. Existing treatment regimens for H. pylori infections have increased failure rates and adverse side effects that desire the search for an effective substitute therapy. Anisochilus carnosus (L.f.) wall (Lamiaceae), a herb which grows once in a year at high elevation is used widely in traditional treatment for the complaints of gastric ulcer and skin diseases. The present study was performed to assess the antibacterial activity of A. carnosus (L.f.) wall, against clinical isolates of H. pylori in vitro.Methods: A. carnosus leaves were collected-dried and extracted with water and ethanol by cold maceration with ethanol by soxhlet method. The minimum inhibitory concentration (MIC) of extracts was made and tested against 32 clinical and 1 reference strains of H. pylori. Results: A. carnosus (L.f.) wall inhibited the growth of most of the clinical H. pylori strains. The MIC of A. carnosus (L.f.) wall extracted by cold maceration (aqueous and ethanol) and Soxhlet apparatus (ethanol) ranged from 500 to 62.5 μg/ml, and the majority of the clinical H. pylori strains were inhibited at the MIC of 500 μg/ml of aqueous, ethanol, and Soxhlet ethanol extraction were 63.63%, 43.75%, and 71.87%, respectively.Conclusion: A. carnosus (L.f.) wall is an efficient inhibitor of H. pylori growth in vitro. A. carnosus (L.f.) wall revealed enormous therapeutic potential to H. pylori infection as it was extremely active in the suppression of H. pylori. Hence, it can be taken as a potential agent against several H. pylori linked gastric pathogenic progressions.

scholarly journals Antimicrobial Activity of Curcumin against Helicobacter pylori Isolates from India and during Infections in Mice

2009 ◽  
Vol 53 (4) ◽  
pp. 1592-1597 ◽  
Author(s):  
Ronita De ◽  
Parag Kundu ◽  
Snehasikta Swarnakar ◽  
T. Ramamurthy ◽  
Abhijit Chowdhury ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Therapeutic Potential ◽  
Alternative Therapy ◽  
Shikimate Pathway ◽  
Antimicrobial Effect ◽  
Gastric Damage ◽  
Shikimate Dehydrogenase ◽  
H Pylori

ABSTRACT Treatment failure is a major cause of concern for the Helicobacter pylori-related gastroduodenal diseases like gastritis, peptic ulcer, and gastric cancer. Curcumin, diferuloylmethane from turmeric, has recently been shown to arrest H. pylori growth. The antibacterial activity of curcumin against 65 clinical isolates of H. pylori in vitro and during protection against H. pylori infection in vivo was examined. The MIC of curcumin ranges from 5 μg/ml to 50 μg/ml, showing its effectiveness in inhibiting H. pylori growth in vitro irrespective of the genetic makeup of the strains. The nucleotide sequences of the aroE genes, encoding shikimate dehydrogenase, against which curcumin seems to act as a noncompetitive inhibitor, from H. pylori strains presenting differential curcumin MICs showed that curcumin-mediated growth inhibition of Indian H. pylori strains may not be always dependent on the shikimate pathway. The antimicrobial effect of curcumin in H. pylori-infected C57BL/6 mice and its efficacy in reducing the gastric damage due to infection were examined histologically. Curcumin showed immense therapeutic potential against H. pylori infection as it was highly effective in eradication of H. pylori from infected mice as well as in restoration of H. pylori-induced gastric damage. This study provides novel insights into the therapeutic effect of curcumin against H. pylori infection, suggesting its potential as an alternative therapy, and opens the way for further studies on identification of novel antimicrobial targets of curcumin.

scholarly journals Flagellar Localization of a Helicobacter pylori Autotransporter Protein

mBio ◽  
2013 ◽  
Vol 4 (2) ◽  
Author(s):  
Jana N. Radin ◽  
Jennifer A. Gaddy ◽  
Christian González-Rivera ◽  
John T. Loh ◽  
Holly M. Scott Algood ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Bacterial Infections ◽  
Secreted Proteins ◽  
Wild Type ◽  
Gram Negative ◽  
Content Type ◽  
Ulcer Disease ◽  
H Pylori ◽  
Type V

ABSTRACTHelicobacter pyloricontains four genes that are predicted to encode proteins secreted by the autotransporter (type V) pathway. One of these, the pore-forming toxin VacA, has been studied in great detail, but thus far there has been very little investigation of three VacA-like proteins. We show here that all three VacA-like proteins are >250 kDa in mass and localized on the surface ofH. pylori. The expression of the threevacA-like genes is upregulated duringH. pyloricolonization of the mouse stomach compared toH. pylorigrowthin vitro, and a wild-typeH. pyloristrain outcompeted each of the three corresponding isogenic mutant strains in its ability to colonize the mouse stomach. One of the VacA-like proteins localizes to a sheath that overlies the flagellar filament and bulb, and therefore, we designate it FaaA (flagella-associated autotransporter A). In comparison to a wild-typeH. pyloristrain, an isogenicfaaAmutant strain exhibits decreased motility, decreased flagellar stability, and an increased proportion of flagella in a nonpolar site. The flagellar localization of FaaA differs markedly from the localization of other known autotransporters, and the current results reveal an important role of FaaA in flagellar localization and motility.IMPORTANCEThe pathogenesis of most bacterial infections is dependent on the actions of secreted proteins, and proteins secreted by the autotransporter pathway constitute the largest family of secreted proteins in pathogenic Gram-negative bacteria. In this study, we analyzed three autotransporter proteins (VacA-like proteins) produced byHelicobacter pylori, a Gram-negative bacterium that colonizes the human stomach and contributes to the pathogenesis of gastric cancer and peptic ulcer disease. We demonstrate that these three proteins each enhance the capacity ofH. pylorito colonize the stomach. Unexpectedly, one of these proteins (FaaA) is localized to a sheath that overliesH. pyloriflagella. The absence of FaaA results in decreasedH. pylorimotility as well as a reduction in flagellar stability and a change in flagellar localization. The atypical localization of FaaA reflects a specialized function of this autotransporter designed to optimizeH. pyloricolonization of the gastric niche.

scholarly journals In vitro antimicrobial activity of Salvadora persica extract on Helicobacter pylori strains isolated from duodenal ulcer biopsies

2012 ◽  
Vol 3 (1) ◽  
pp. 9 ◽  
Author(s):  
Ehsan Mirkamandar ◽  
Mohammad Reza Shakibaie ◽  
Saeed Adeli ◽  
Mitra Mehrabani ◽  
Mohammad Mehdi Hayatbakhsh ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Duodenal Ulcer ◽  
Helicobacter Pylori ◽  
Dilution Method ◽  
Minimal Bactericidal Concentration ◽  
Agar Dilution Method ◽  
Zone Of Inhibition ◽  
Salvadora Persica ◽  
H Pylori

The aim of this study was to evaluate the <em>in vitro</em> antimicrobial activity of a methanolic extract of <em>Salvadora persica</em> solution on <em>Helicobacter pylori</em> isolated from duodenal ulcer. Over 22 strains of H. pylori were isolated from duodenal ulcer from August 2010 to June 2011. The <em>S. persica</em> stem was purchased from a local herb market and finely powdered. Extraction was performed with 60% methanol using a soxhlet extractor for 48 h until the solvent turned colorless while being incubated in an oven at 40°C for 48 h till dried. Dry powder was used to determine antimicrobial activity by the agar ditch method. Minimum inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of the extract were determined by the agar dilution method. At concentrations of 10, 100, 200, 500 µg/mL, no zone of inhibition around the ditches was observed while a clear zone of inhibition (12 mm) was detected at 1000 µg/mL concentration for all the isolates. The best antimicrobial activity was observed at MIC 1000 µg/mL (P≤0.05). Furthermore, 10 out of 22 isolates were inhibited at 750 µg/mL of the extract. The MBC results showed that at a concentration of 1000 µg/mL all cells were dead while at a concentration of 750 µg/mL of<em> S. persica </em>a few <em>H. pylori</em> cells were still able to form colonies on Brucella agar supplemented with sheep red blood cells and antibiotics. From the above results it can be concluded that high concentration of S.persica could inhibit the growth of H. pylori and MIC and MBC were similar at that concentration.

scholarly journals The Lipid A 1-Phosphatase of Helicobacter pylori Is Required for Resistance to the Antimicrobial Peptide Polymyxin

2006 ◽  
Vol 188 (12) ◽  
pp. 4531-4541 ◽  
Author(s):  
An X. Tran ◽  
Judy D. Whittimore ◽  
Priscilla B. Wyrick ◽  
Sara C. McGrath ◽  
Robert J. Cotter ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Antimicrobial Peptide ◽  
Lipid A ◽  
Mass Spectrometry Analysis ◽  
Cationic Antimicrobial Peptide ◽  
Vitro Assay ◽  
Chloramphenicol Resistance ◽  
Gram Negative ◽  
H Pylori

ABSTRACT Modification of the phosphate groups of lipid A with amine-containing substituents, such as phosphoethanolamine, reduces the overall net negative charge of gram-negative bacterial lipopolysaccharide, thereby lowering its affinity to cationic antimicrobial peptides. Modification of the 1 position of Helicobacter pylori lipid A is a two-step process involving the removal of the 1-phosphate group by a lipid A phosphatase, LpxEHP (Hp0021), followed by the addition of a phosphoethanolamine residue catalyzed by EptAHP (Hp0022). To demonstrate the importance of modifying the 1 position of H. pylori lipid A, we generated LpxEHP-deficient mutants in various H. pylori strains by insertion of a chloramphenicol resistance cassette into lpxEHP and examined the significance of LpxE with respect to cationic antimicrobial peptide resistance. Using both mass spectrometry analysis and an in vitro assay system, we showed that the loss of LpxEHP activity in various H. pylori strains resulted in the loss of modification of the 1 position of H. pylori lipid A, thus confirming the function of LpxEHP. Due to its unique lipid A structure, H. pylori is highly resistant to the antimicrobial peptide polymyxin (MIC > 250 μg/ml). However, disruption of lpxEHP in H. pylori results in a dramatic decrease in polymyxin resistance (MIC, 10 μg/ml). In conclusion, we have characterized the first gram-negative LpxE-deficient mutant and have shown the importance of modifying the 1 position of H. pylori lipid A for resistance to polymyxin.

scholarly journals Engineered endolysin-based “artilysins” for controlling the gram-negative pathogen Helicobacter pylori

AMB Express ◽  
2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Dengyuan Xu ◽  
Shanshan Zhao ◽  
Jun Dou ◽  
Xiaofeng Xu ◽  
Yanyan Zhi ◽  
...  
Keyword(s):  
Cell Wall ◽  
Helicobacter Pylori ◽  
Transmembrane Protein ◽  
Gastrointestinal Diseases ◽  
Amide Bond ◽  
Resistant Bacteria ◽  
Gram Negative ◽  
Antibiotic Overuse ◽  
H Pylori

AbstractHelicobacter pylori infection can cause a variety of gastrointestinal diseases. In severe cases, there is a risk of gastric cancer. Antibiotics are often used for clinical treatment of H. pylori infections. However, because of antibiotic overuse in recent years and the emergence of multidrug-resistant bacteria, there is an urgent need to develop new treatment methods and drugs to achieve complete eradication of H. pylori. Endolysins and holins encoded by bacterial viruses (i.e., phages) represent a promising avenue of investigation. These lyase-based antibacterial drugs act on the bacterial cell wall to destroy the bacteria. Currently, a type of endolysin that has been studied more frequently acts on the amide bond between peptidoglycans, and holin is a transmembrane protein that can punch holes in the cell membrane. However, as a Gram-negative bacterium, H. pylori possesses a layer of impermeable lipopolysaccharides on the cell wall, which prevents endolysin interaction with the cell wall. Therefore, we designed a genetic linkage between an endolysin enzyme and a holin enzyme with a section of polypeptides (e.g., polycations and hydrophobic peptides) that enable penetration of the outer membrane. These complexes were designated “artilysins” and were efficiently expressed in Escherichia coli. In vitro bacteriostasis experiments showed that the purified artilysins had strong bacteriostatic effects on H. pylori. In addition, the surface of H. pylori was perforated and destroyed, as confirmed by electron microscopy, which was proved that artilysins had bacteriolytic effect on H. pylori.

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