The cultivation of Acanthamoeba using with different axenic and monoxenic media

Eroglu, Fadime; Evyapan, Gülşah; Koltas, Ismail Soner

doi:10.19127/mbsjohs.71412

Cited by 11 publications

(7 citation statements)

References 18 publications

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“…The results of the present study were consistent with the results of Eroğlu; et al., they observed the highest growth peak in RPMI-FBS, however, this finding agrees with the increase in growth in the other culture media. Also, there were no signficant differences in growth in PYG culture medium as compared to our study ( Eroğlu et al., 2015 ). In another study by Peretz, it was observed that culture in PYG and TYI-S-33 medium could help to diagnose Acanthamoe ba keratitis alongside the calcofluor white staining and fluorescence microscopy.…”

Section: Discussioncontrasting

confidence: 67%

Growth comparison of Acanthamoeba genotypes T3 and T4 in several culture media

Latifi

Salimi

2020

Heliyon

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Acanthamoeba causes severe diseases such as Granulomatous Amebic Encephalitis (GAE) and Acanthamoeba keratitis (AK). Improving the culture media classically used for this amoeba could help to identify it quickly and facilitate its study as a biological model. The purpose of this study was to compare the growth of two Acanthamoeba genotypes (T3 and T4) in several culture media. Acanthamoeba griffini (T3 genotype) and Acanthamoeba castellanii (T4 genotype) were cultured in PYG, TSY, TYI-S-33, RPMI, and RPMI-FBS medium. The number of amoebas grown in different culture media was counted and compared to each other for 14 days. Findings in this research revealed the highest growth in RPMI-FBS medium. For this reason, we can recommend this culture medium to promote the growth of Acanthamoeba in its biological studies.

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Section: Discussioncontrasting

confidence: 67%

Growth comparison of Acanthamoeba genotypes T3 and T4 in several culture media

Latifi

Salimi

2020

Heliyon

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“…A wide range of bacteria have been used in co-culture with Acanthamoeba. The most common microbes used to culture Acanthamoeba are E. coli , Klebsiella aerogenes [ 88 , 89 , 90 ] and Enterobacter spp. (E. cloacae and E. aerogenes ) [ 8 , 25 , 59 ] on NNA or in saline [ 83 ] ( Figure 3 ).…”

Section: Resultsmentioning

confidence: 99%

A Systematic Review of Intracellular Microorganisms within Acanthamoeba to Understand Potential Impact for Infection

et al. 2021

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Acanthamoeba, an opportunistic pathogen is known to cause an infection of the cornea, central nervous system, and skin. Acanthamoeba feeds different microorganisms, including potentially pathogenic prokaryotes; some of microbes have developed ways of surviving intracellularly and this may mean that Acanthamoeba acts as incubator of important pathogens. A systematic review of the literature was performed in order to capture a comprehensive picture of the variety of microbial species identified within Acanthamoeba following the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines. Forty-three studies met the inclusion criteria, 26 studies (60.5%) examined environmental samples, eight (18.6%) studies examined clinical specimens, and another nine (20.9%) studies analysed both types of samples. Polymerase chain reaction (PCR) followed by gene sequencing was the most common technique used to identify the intracellular microorganisms. Important pathogenic bacteria, such as E. coli, Mycobacterium spp. and P. aeruginosa, were observed in clinical isolates of Acanthamoeba, whereas Legionella, adenovirus, mimivirus, and unidentified bacteria (Candidatus) were often identified in environmental Acanthamoeba. Increasing resistance of Acanthamoeba associated intracellular pathogens to antimicrobials is an increased risk to public health. Molecular-based future studies are needed in order to assess the microbiome residing in Acanthamoeba, as a research on the hypotheses that intracellular microbes can affect the pathogenicity of Acanthamoeba infections.

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“…The genome sequences of the isolates were submitted to GenBank under the accession numbers MH791020, MH790983 and MN700296 ( Mohd Hussain et al, 2019 , Mohd Hussain et al, 2020 ). Trophozoites were grown monoxenically on a new 1.5% non-nutrient agar (NNA) (Sigma Aldrich A7002, USA) plates seeded with E. coli at 30 ⁰C for three days ( Eroğlu et al, 2015 ). The preparation of cysts was made from approximately 100 late-log-phase trophozoite cultures using 1.5% NNA medium, as previously established by de Aguiar et al (2013) .…”

Section: Methodsmentioning

confidence: 99%

In vitro effects of multi-purpose contact lens disinfecting solutions towards survivability of Acanthamoeba genotype T4 in Malaysia

Hussain

Afiqah

Ghani

et al. 2021

Saudi Journal of Biological Sciences

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The incidence of Acanthamoeba keratitis has been increasing since the previous decades, especially among contact lens users. This infection is majorly caused by the use of ineffective contact lens disinfecting solution. Thus, this study was conducted to evaluate the in vitro effects of multi-purpose disinfecting solutions (MPDS) against Acanthamoeba trophozoites and cysts. Acanthamoeba genotype T4 isolated from contact lens paraphernalia and an environmental strains were propagated for trophozoite or cyst-containing culture and adjusted in final concentration of 1 × 10 5 cells/ml. Amoebicidal and cysticidal assays were conducted by incubating trophozoites and cysts with OPTI-FREE® Express®, ReNu® Fresh™, Complete® Multi-Purpose Solution and AVIZOR Unica® Sensitive according to the manufacturer’s minimum recommended disinfectant time (MMRDT) for up to 12 h at 30 ⁰C. Trypan blue hemocytometer-based microscopic counts determined amoebicidal and cysticidal effects. The viability of Acanthamoeba trophozoites and cysts was confirmed by re-inoculated them in the 1.5% non-nutrient agar plates. It was found that none of the MPDS showed amoebicidal and cysticidal effects during the MMRDT. However, OPTI-FREE® Express® demonstrated a significant differences in average cell reduction for both stages within MMRDT. When subjected to 12 h exposure, both OPTI-FREE® Express® and ReNu® Fresh™ led to significant reduction in the number of trophozoite and cyst cells. Notably, Complete® Multi-Purpose Solution and AVIZOR Unica® Sensitive did appreciably improve the solution effectiveness towards trophozoite cells when incubated for 12 h. All MPDS were largely ineffective, with 100% survival of all isolates at MMRDT, while OPTI-FREE® Express® showed limited amoebicidal activity against the contact lens paraphernalia isolate, however, it was more against the environmental strains after 12 h incubation time. The commercially available MPDS employed in this research offered minimal effectiveness against the protozoa despite the contact time. Improvement or development of new solution should consider the adjustment of the appropriate disinfectant concentration, adequate exposure time or the incorporation of novel chemical elements, which are effective against Acanthamoeba for accelerated disinfecting and more reduction of potential exposure of contact lens users to Acanthamoeba keratitis.

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The cultivation of Acanthamoeba using with different axenic and monoxenic media

Cited by 11 publications

References 18 publications

Growth comparison of Acanthamoeba genotypes T3 and T4 in several culture media

Growth comparison of Acanthamoeba genotypes T3 and T4 in several culture media

A Systematic Review of Intracellular Microorganisms within Acanthamoeba to Understand Potential Impact for Infection

In vitro effects of multi-purpose contact lens disinfecting solutions towards survivability of Acanthamoeba genotype T4 in Malaysia

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