Diagnosis of Balamuthia mandrillaris Encephalitis by Thymine–Adenine Cloning Using Universal Eukaryotic Primers

Kim, Ju Yeong; Yi, Myung Hee; Kim, Myung-Jun; Yeom, Joon‐Sup; Yoo, Hyun Dong; Kim, Sang Woo; Yong, Tai Soon

doi:10.3343/alm.2022.42.2.196

Cited by 2 publications

(2 citation statements)

References 25 publications

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“…Polymerase chain reaction (PCR) was performed to identify the amoebae isolates using reaction systems containing 2 μg of template DNA, 0.5 μl of Taq buffer, 3 μl of 10× buffer, 2 μl of dNTPs, and 2 μl each of the forward and reverse 18S rDNA primers (V4-1F 5′-GCGGTAATTCCAGCTC-3′ and V4-4R 5′-GCCMTTCCGTCAATTCC-3′) [ 21 ]. The amplifications were performed with forward and reverse 16s rDNA primers (27F 5′-AGAGTTTGATCCTGGCTCAG-3′ and 1492R 5′-GGTTACCTTGTTACGACTT-3′) to identify the bacterial endosymbiont, and the total volume was adjusted to 30 μl using DW.…”

Section: Methodsmentioning

confidence: 99%

Presence and diversity of free-living amoebae and their potential application as water quality indicators

Choi,

Seong,

Kim

et al. 2024

Parasites Hosts Dis

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Free-living amoebae (FLA) are found in diverse environments, such as soils, rivers, and seas. Hence, they can be used as bioindicators to assess the water quality based solely on their presence. In this study, we determined the presence of FLA in river water by filtering water samples collected from various sites and culturing the resulting filtrates. FLA were detected in all the water samples with varying quality grades (Grades Ι-V). The significant increase in the size of the amoebae population with the deterioration in the water quality. Monoxenic cultures of the amoebae were performed, and genomic DNAs were isolated, among which 18S rDNAs were sequenced to identify the amoeba species. Of the 12 species identified, 10 belonged to the Acanthamoeba genus; of the remaining 2 species, one was identified as Vannella croatica and the other as a species of Vermamoeba. Acanthamoeba was detected in samples with Grades Ι to VI quality, whereas the Vermamoeba species was present only in Grade Ι water. V. croatica was found exclusively in water with Grade ΙΙ quality. Following morphological observations, genomic DNA was sequenced using 16S rDNA to determine whether the species of Acanthamoeba harbored endosymbionts. Most of the isolated Acanthamoeba contained endosymbionts, among which 4 species of endogenous bacteria were identified and examined using transmission electron microscopy. This study provides evidence that the distribution of amoebae other than Acanthamoeba may be associated with water quality. However, further confirmation will be required based on accurate water quality ratings and assessments using a more diverse range of FLA.

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Section: Methodsmentioning

confidence: 99%

Presence and diversity of free-living amoebae and their potential application as water quality indicators

Choi,

Seong,

Kim

et al. 2024

Parasites Hosts Dis

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“…Naegleria fowleri and Balamuthia mandrillaris are free-living, eukaryotic, unicellular amoebae that pose a major risk towards human health [ 1 , 2 , 3 ]. These amoebae are found across the environment globally [ 4 ].…”

Section: Introductionmentioning

confidence: 99%

Zinc Oxide Nanoconjugates against Brain-Eating Amoebae

Siddiqui

Boghossian²,

Akbar³

et al. 2022

Antibiotics

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Naegleria fowleri and Balamuthia mandrillaris are opportunistic protists, responsible for fatal central nervous system infections such as primary amoebic meningoencephalitis (PAM) and granulomatous amoebic encephalitis (GAE) with mortality rates higher than 90%. Threatening a rise in cases is the increase in temperature due to global warming. No effective treatment is currently available. Herein, nanotechnology was used to conjugate Zinc oxide with Ampicillin, Ceftrixon, Naringin, Amphotericin B, and Quericitin, and the amoebicidal activity and host cell cytotoxicity of these resulting compounds were investigated. The compounds ZnO-CD-AMPi, ZnO-CD-CFT, ZnO-CD-Nar, ZnO-CD-AMB, and ZnO-CD-QT were found to reduce N. fowleri viability to 35.5%, 39.6%, 52.0%, 50.8%, 35.9%, and 69.9%, respectively, and B. mandrillaris viability to 40.9%, 48.2%, 51.6%, 43.8%, and 62.4%, respectively, when compared with their corresponding controls. Furthermore, the compounds reduced N. fowleri-mediated and B. mandrillaris-mediated host cell death significantly. Additionally, the compounds showed limited cytotoxicity against human cells; cell toxicity was 35.5%, 36.4%, 30.9%, 36.6%, and 35.6%, respectively, for the compounds ZnO-CD-AMPi, ZnO-CD-CFT, ZnO-CD-Nar, ZnO-CD-AMB, and ZnO-CD-QT. Furthermore, the minimum inhibitory concentrations to inhibit amoeba growth by 50% were determined for N. fowleri and B. mandrillaris. The MIC50 for N. fowleri were determined to be 69.52 µg/mL, 82.05 µg/mL, 88.16 µg/mL, 95.61 µg/mL, and 85.69 µg/mL, respectively; the MIC50 of the compounds for B. mandrillaris were determined to be 113.9 µg/mL, 102.3 µg/mL, 106.9 µg/mL, 146.4 µg/mL, and 129.6 µg/mL, respectively. Translational research to further develop therapies based on these compounds is urgently warranted, given the lack of effective therapies currently available against these devastating infections.

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Diagnosis of Balamuthia mandrillaris Encephalitis by Thymine–Adenine Cloning Using Universal Eukaryotic Primers

Cited by 2 publications

References 25 publications

Presence and diversity of free-living amoebae and their potential application as water quality indicators

Presence and diversity of free-living amoebae and their potential application as water quality indicators

Zinc Oxide Nanoconjugates against Brain-Eating Amoebae

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