scholarly journals Cultivation of Entamoeba histolytica in vitro and diagnose the bacterial growths in culture media

2009 ◽  
Vol 6 (3) ◽  
pp. 442-447
Author(s):  
Baghdad Science Journal
Keyword(s):  
Escherichia Coli ◽  
Entamoeba Histolytica ◽  
Culture Media ◽  
Agar Medium ◽  
Cyst Formation ◽  
Essential Requirement ◽  
Bacteria Growth ◽  
Bacterial Content ◽  
Infusion Agar

The parasite was isolated from a stool sample, cultivated and maintained in vitro using Locke-egg medium (LEM) and Liver infusion agar medium (LIAM) . The culture was maintained for up to 21 months, and the best time to maintain the parasite was every 48 hours, although the growth in the culture media continued for 13 days without a maintenance. Additionally, no cyst formation was observed during cultivation of parasite in the two culture media. Although, was observe young cyst formed in LEM media were deletion of maintained. The diagnosis of bacteria growth in the culture media, bacterial content (Escherichia coli) was an dominance and essential requirement for a successful cultivation of Entamoeba histolytica in the two culture media.

scholarly journals The effect of calcium antagonists on the growth of Entamoeba histolytica in vitro

2010 ◽  
Vol 4 (1) ◽  
pp. 5-10
Author(s):  
Zahra’a Abdul-Raheem Ahmed ◽  
Ali H. Ad’hiah ◽  
Amna N. Jasim
Keyword(s):  
Entamoeba Histolytica ◽  
Calcium Antagonists ◽  
Culture Media ◽  
Agar Medium ◽  
Reproduction Rate ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Infusion Agar ◽  
Ethylene Diaminetetraacetic Acid

he E. histolytica parasite was maintained in vitro using Locke-egg medium (LEM) and Liver infusion agar medium (LIAM). The effect of two calcium antagonists (Nifedipine and Ethylene-diaminetetraacetic acid EDTA) on the growth and activity of the parasite in the two culture media was investigated. The calcium antagonists Nifedipine and EDTA inhibited the reproduction rate of E. histolytica in a concentration-dependent manner. For Nifedipine, a concentration of 41.6 mg/ml inhibited the reproduction rate to 99.7% in both media. The EDTA had an approximate effect (98.2 and 95.8)% at a concentration of 0.83 mg/ml in LEM and LIAM media, respectively. Additionally, some cases of a parasite encystment were observed in LEM medium that was treated with Nifedipine.

scholarly journals The effect of aqueous plant extracts (Hibiscus sabdariffa and Glycyrrhiza glabra), on the growth of Entamoeba histolytica in vitro

2009 ◽  
Vol 3 (1) ◽  
pp. 50-55
Author(s):  
Zahra’a Abdul-Raheem Ahmed ◽  
Amna N. Jasim ◽  
Ali H. Ad’hiah
Keyword(s):  
Entamoeba Histolytica ◽  
Plant Extracts ◽  
Culture Media ◽  
Agar Medium ◽  
Glycyrrhiza Glabra ◽  
Reproduction Rate ◽  
Hibiscus Sabdariffa ◽  
Significant Difference ◽  
Infusion Agar

Entamoeba histolytica parasite was isolated from a stool sample, cultivated and maintained in vitro using Locke-egg medium (LEM) and Liver infusion agar medium (LIAM). The effect of two aqueous plant extracts (Hibiscus sabdariffa and Glycyrrhiza glabra) on the growth and activity of the parasite in the two culture media was investigated. The aqueous extracts of H. sabdariffa and G. glabra were effective in reducing the parasite size in the LEM medium. With respect to the reproduction rate, the third concentration (19.71 mg/ml) of H. sabdariffa was significantly effective in inhibiting such rate to 57.6 and 83.6% in LEM and LIAM media, respectively, while for G. glabra, no significant difference in the reproduction rate was observed in both culture media.

scholarly journals The role of some types of erythrocytes on the growth of Entamoeba histolytica trophozoite in vitro

2010 ◽  
Vol 7 (1) ◽  
pp. 200-206
Author(s):  
Baghdad Science Journal
Keyword(s):  
Stool Sample ◽  
Entamoeba Histolytica ◽  
Culture Media ◽  
Agar Medium ◽  
Reproduction Rate ◽  
Sheep Erythrocytes ◽  
Histolytica Trophozoite ◽  
Infusion Agar

The parasite E.histolytica was first isolated from a stool sample, and then cultivated and maintained in vitro using Locke-egg medium (LEM) and Liver infusion agar medium (LIAM) . Then, the effect of some types of erythrocytes (human and sheep), on the growth and activity of the parasite in the two culture media was investigated. The parasite was able to ingest and lysis erythrocytes of human and sheep that were supplemented to the culture media and such manipulation was able to augment the reproduction rate of the cultivated E. histolytica, however, such consequence was media- and concentration-dependent. The reproduction rate was significantly increased (66.0, 57.5 and 58.6%, respectively) in LEM medium containing human erythrocytes types B at 0.11 x 106 cells/ml and O at 0.13 x 106 and 0.15 x 106 cells/ml. The sheep erythrocytes showed a similar enhancement (56.1%) at a concentration of 0.13 x 106 cells/ml. In contrast, adding erythrocytes to LIAM medium did not enhance the reproduction rate of the parasite significantly.

scholarly journals Impact of Different Carbon Sources on the in vitro Growth and Viability of Escherichia coli (SUBE01) and Salmonella spp. (SUBS01) Cells

2016 ◽  
pp. 39-44
Author(s):  
Ifra Tun Nur ◽  
Jannatun Tahera ◽  
Md Sakil Munna ◽  
M Majibur Rahman ◽  
Rashed Noor
Keyword(s):  
Escherichia Coli ◽  
Bacterial Growth ◽  
Culture Media ◽  
Bacterial Species ◽  
Carbon Sources ◽  
Growth Dynamics ◽  
Luria Bertani ◽  
Tween 20 ◽  
Salmonella Spp

With a previous observation of Escherichia coli growth cessation along with temperature variation within three different bacteriological culture media (nutrient agar, Luria-Bertani agar and minimal agar), current investigation further depicted on the possible growth dynamics of Escherichia coli (SUBE01) and Salmonella (SUBS01) growth and viability upon supplementation of different carbon sources (dextrose, sucrose, lactose, glycerol and tween 20) at 37°C under the aeration of 100 rpm. Viability of the tested bacterial species was assessed through the enumeration of the colony forming unit (cfu) appeared upon prescribed incubation for 12-24 hours on different agar plates consisting of the above mentioned carbon sources. Besides, to inspect the cellular phenotypic changes, morphological observations were conducted under the light microscope. Variations in bacterial growth (either growth acceleration or cessation) were further noticed through the spot tests on the agar plates. Considerable shortfalls in the culturable cells of E. coli and Salmonella spp. were noted in the minimal media separately consisting of sucrose, lactose, glycerol or tween 20 while an opposite impact of accelerated growth was noticed in the media supplied with dextrose. The data revealed a hierarchy of consequence of carbon sources as nutrient generators whereby the favourable bacterial growth and survival order of the carbon sources was estimated as dextrose > glycerol > lactose > tween 20 > sucrose.Bangladesh J Microbiol, Volume 32, Number 1-2,June-Dec 2015, pp 39-44

Persistent bloody diarrhoea without fever associated with diffusely adherent Escherichia coli in a young child

2013 ◽  
Vol 62 (12) ◽  
pp. 1907-1910 ◽  
Author(s):  
Sandra Patzi-Vargas ◽  
Mussaret Zaidi ◽  
Rodolfo Bernal-Reynaga ◽  
Magda León-Cen ◽  
Alba Michel ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Culture Media ◽  
Evidence Base ◽  
Antimicrobial Treatment ◽  
Molecular Techniques ◽  
Bloody Diarrhoea ◽  
Severe Diarrhoea ◽  
E Coli ◽  
Clinical Laboratories

Diffusely adherent Escherichia coli (DAEC) is thought to cause diarrhoea in children, and so too are other diarrhoeagenic E. coli (DEC); however, the evidence base is inconclusive. DEC pathotypes are differentiated on the basis of their pathogenic features, and thus cannot be quickly identified on selective culture media. Molecular techniques, not readily available in most clinical laboratories, are required to differentiate DEC strains from non-pathogenic E. coli in the stool flora. We report a case of persistent bloody diarrhoea, without fever, in a previously healthy 21-month infant from whom we isolated five DAEC strains. The child’s stools movements were loose, with gross blood and mucus; fresh mount analysis revealed numerous faecal leukocytes and erythrocytes. Response to antimicrobial treatment with trimethoprim-sulfamethoxazole was poor despite susceptibility in vitro. Although the patient improved with azithromycin, blood was present in the patient’s stools for over 30 days. The severe diarrhoea in this patient might be explained by the fact that these DAEC isolates harboured a siderophore receptor, which allows the bacteria to use iron derived from haem compounds that promote its multiplication. The isolates also induced in vitro secretion of several immunomodulatory cytokines that may account for the patient’s loose stools and faecal leukocytes. DAEC may play a greater role than suspected in afebrile children with bloody diarrhoea.

scholarly journals In Vitro Evaluation of Dietary Fiber Anti-Infectious Properties against Food-Borne Enterotoxigenic Escherichia coli

Nutrients ◽  
2021 ◽  
Vol 13 (9) ◽  
pp. 3188
Author(s):  
Thomas Sauvaitre ◽  
Claude Durif ◽  
Adeline Sivignon ◽  
Sandrine Chalancon ◽  
Tom Van de Wiele ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Culture Media ◽  
Toxin Production ◽  
Enterotoxigenic Escherichia Coli ◽  
In Vitro Assays ◽  
Dietary Fibers ◽  
Mucus Cell ◽  
Etec Strain ◽  
Beneficial Effects

Dietary fibers have well-known beneficial effects on human health, but their anti-infectious properties against human enteric pathogens have been poorly investigated. Enterotoxigenic Escherichia coli (ETEC) is the main agent of travelers’ diarrhea, against which targeted preventive strategies are currently lacking. ETEC pathogenesis relies on multiple virulence factors allowing interactions with the intestinal mucosal layer and toxins triggering the onset of diarrheal symptoms. Here, we used complementary in vitro assays to study the antagonistic properties of eight fiber-containing products from cereals, legumes or microbes against the prototypical human ETEC strain H10407. Inhibitory effects of these products on the pathogen were tested through growth, toxin production and mucus/cell adhesion inhibition assays. None of the tested compounds inhibited ETEC strain H10407 growth, while lentil extract was able to decrease heat labile toxin (LT) concentration in culture media. Lentil extract and specific yeast cell walls also interfered with ETEC strain H10407 adhesion to mucin beads and human intestinal cells. These results constitute a first step in the use of dietary fibers as a nutritional strategy to prevent ETEC infection. Further work will be dedicated to the study of fiber/ETEC interactions within a complex gut microbial background.

scholarly journals Antimicrobial Activity of Nisin and Lysozyme on Foodborne Pathogens Listeria Monocytogenes, Staphylococcus Aureus, Salmonella Typhimurium, and Escherichia Coli at Different pH

2018 ◽  
Author(s):  
Hamdollah Moshtaghi ◽  
Azadeh Rashidimehr ◽  
Behzad Shareghi
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Listeria Monocytogenes ◽  
Foodborne Pathogens ◽  
Inhibitory Concentration ◽  
Low Ph ◽  
Bacteria Growth ◽  
Microdilution Method ◽  
Natural Preservatives

Background: To prevent or inhibit the growth of pathogenic microorganisms and food spoilage factors, many studies have been done by using natural preservatives. The aim of study was to investigate the effect of different concentrations of lysozyme and Nisin on the growth rate and also to determine the minimum inhibitory concentration (MIC) and minimum bactericidal cocentratiin (MBC) of these combinations on the bacteria of Escherichia coli, Staphylococcus aureus, Salmonella typhimorium and Listeria monocytogenes. Methods: In this study, various concentrations of lysozyme and Nisin were set in form of alone concentration and in combination concentrations (0, 19.53, 39.06, 78.13, 156.25, 312.5, 625, 1250, 2500, 5000) in vitro conditions and 6 pH 5.5, 6, 6.5, 7, 7.5, and 8. Microdilution method at 24°C was done and the combined effect on bacteria growth was read by using ELISA reader. Results: The results showed that lysozyme was less effective on Escherichia coli and Nisin was less effective on Listeria monocytogenes. Moreover, combining lysozyme and Nisin at low pH decreased the MIC. Conclusions: The results of the study showed that the effect of combining lysozyme and Nisin on Staphylococcus aureus is above all other bacteria and at low pH reduces the MIC.

scholarly journals Impact of thrombocytes, on bacterial growth and antimicrobial activity of selected antibiotics

2019 ◽  
Vol 39 (3) ◽  
pp. 593-597 ◽  
Author(s):  
Alina Karoline Nussbaumer-Pröll ◽  
Sabine Eberl ◽  
Birgit Reiter ◽  
Thomas Stimpfl ◽  
Walter Jäger ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Bacterial Growth ◽  
Limiting Factor ◽  
Growth Media ◽  
Healthy Human ◽  
Bacterial Killing ◽  
Bacteria Growth ◽  
The Impact

AbstractIn vitro pharmacodynamic models are used to optimize in vivo dosing regimens in antimicrobial drug development. One limiting factor of such models is the lack of host factors such as corpuscular blood components as erythrocytes which have already been shown to impact activity of antibiotics and/or growth of the pathogen. However, the impact of thrombocytes has not previously been investigated. We set out to investigate if the addition of thrombocytes (set to physiological concentrations in blood of healthy human, i.e., 5 × 105 thrombocytes/μL standard growth media Mueller Hinton Broth, MHB) has an influence on bacterial growth and on the efficacy of antibiotics against Gram+ and Gram− bacteria. Growth assays and time-killing-curves (TKC) were performed with ATCC-strains of Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa in triplicate over 24 h. The same approach was followed for 5 clinical isolates of Escherichia coli. Meropenem, ciprofloxacin, and tigecycline were tested as representatives of broad-spectrum antibiotics, and concentrations several-fold above and below the minimal inhibitory concentration (MIC) were simulated. No significant impact of thrombocytes was found on bacterial growth or antimicrobial stability for the investigated agents. Bacteria reduced thrombocyte content to different degree, indicating direct interaction of pathogens and thrombocytes. Impact on bacterial killing was observed but was not fully reproducible when thrombocytes from different donors where used. While interaction of bacteria and thrombocytes was evident in the present study, interaction between antibiotic activity and thrombocytes seems unlikely. Whether variability was caused by different thrombocyte concentrates needs further investigation.

scholarly journals Entamoeba histolytica Interaction with Enteropathogenic Escherichia coli Increases Parasite Virulence and Inflammation in Amebiasis

2019 ◽  
Vol 87 (12) ◽  
Author(s):  
Luz A. Fernández-López ◽  
Karla Gil-Becerril ◽  
Silvia Galindo-Gómez ◽  
Teresa Estrada-García ◽  
Cecilia Ximénez ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Entamoeba Histolytica ◽  
Proinflammatory Cytokine ◽  
Fold Increase ◽  
Symptomatic Infection ◽  
Content Type ◽  
Inflammatory Reactions ◽  
Cytokine Genes ◽  
E Coli

ABSTRACT Epidemiological studies suggest frequent association of enteropathogenic bacteria with Entamoeba histolytica during symptomatic infection. In this study, we sought to determine if the interaction with enteropathogenic (EPEC) or nonpathogenic Escherichia coli (strain DH5α) could modify the virulence of E. histolytica to cause disease in animal models of amebiasis. In vitro studies showed a 2-fold increase in CaCo2 monolayer destruction when E. histolytica interacted with EPEC but not with E. coli DH5α for 2.5 h. This was associated with increased E. histolytica proteolytic activity as revealed by zymogram analysis and degradation of the E. histolytica CP-A1/5 (EhCP-A1/5) peptide substrate Z-Arg-Arg-pNC and EhCP4 substrate Z-Val-Val-Arg-AMC. Additionally, E. histolytica-EPEC interaction increased EhCP-A1, -A2, -A4, and -A5, Hgl, Apa, and Cox-1 mRNA expression. Despite the marked upregulation of E. histolytica virulence factors, nonsignificant macroscopic differences in amebic liver abscess development were observed at early stages in hamsters inoculated with either E. histolytica-EPEC or E. histolytica-E. coli DH5α. Histopathology of livers of E. histolytica-EPEC-inoculated animals revealed foci of acute inflammation 3 h postinoculation that progressively increased, producing large inflammatory reactions, ischemia, and necrosis with high expression of il-1β, ifn-γ, and tnf-α proinflammatory cytokine genes compared with that in livers of E. histolytica-E. coli DH5α-inoculated animals. In closed colonic loops from mice, intense inflammation was observed with E. histolytica-EPEC manifested by downregulation of Math1 mRNA with a corresponding increase in the expression of Muc2 mucin and proinflammatory cytokine genes il-6, il-12, and mcp-1. These results demonstrate that E. histolytica/EPEC interaction enhanced the expression and production of key molecules associated with E. histolytica virulence, critical in pathogenesis and progression of disease.

scholarly journals Expression of a Mycobacterium tuberculosis Arabinomannan Antigen In Vitro and In Vivo

2001 ◽  
Vol 69 (9) ◽  
pp. 5671-5678 ◽  
Author(s):  
J. Reid Schwebach ◽  
Arturo Casadevall ◽  
Rachel Schneerson ◽  
Zhongdong Dai ◽  
Xiaojuan Wang ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Culture Media ◽  
Extended Period ◽  
Tween 80 ◽  
Surface Expression ◽  
Dependent Manner ◽  
Bacteria Growth ◽  
Growth Changes

ABSTRACT The outermost layer of Mycobacterium tuberculosiscontains two major polysaccharides, arabinomannan (AM) and glucan (GC). We studied the in vitro and in vivo expression of anM. tuberculosis AM antigen using monoclonal antibody (MAb) 9d8 (2a), an isotype-switched variant of the immunoglobulin G3 (IgG3) MAb 9d8. MAb 9d8 had been previously shown to bind M. tuberculosis AM and the M. tuberculosis surface. Our in vitro experiments showed that MAb 9d8(2a) bound strongly to whole-cell M. tuberculosis Erdman but not to the CDC 1551 strain grown in medium for an extended period. However, AM antigen was detected in the culture supernatant of both strains, and its concentration increased in a time-dependent manner. The detection of AM antigen from both strains was decreased in the presence of Tween 80. In mice infected with M. tuberculosis Erdman, AM antigen accumulated in organ homogenates concomitant to an increase in bacterial organ burden and an increase in IgG and IgM titer to AM. These results (i) indicate that the surface expression of AM during in vitro growth changes with culture age, is strain dependent, and is affected by the presence of Tween 80 in the culture media; (ii) show that AM is produced by bacteria growth in vivo; and (iii) demonstrate that the amount of in vivo-detected AM can be dependent on the number of bacteria in the infected organ.

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