Molecular Identification of Pathogenic Free-Living Amoeba from Household Biofilm Samples in Iran: A Risk Factor for Acanthamoeba Keratitis

Norouzi, Maryam; Saberi, Reza; Niyyati, Maryam; Lorenzo‐Morales, Jacob; Mirjalali, Hamed; Fatemi, Marziye; Javanmard, Ehsan; Karamati, Seyed Ahmad

doi:10.3390/microorganisms9102098

Cited by 10 publications

(5 citation statements)

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“…Recent study has reported that Vermamoeba (Hartmannella) vermiformis as the etiological agent of an infection close to the eye of a female patient [5]. Some investigators checked localization of FLAs in our environments because they thought that FLA is a potential pathogen or vector of bacterial pathogens [33][34][35][36]. FLAs were detected with potential human pathogenic bacteria in stored water in rural households in South Africa [34].…”

Section: Discussionmentioning

confidence: 99%

Free-Living Amoeba Vermamoeba vermiformis Induces Allergic Airway Inflammation

et al. 2022

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The high percentage of Vermamoeba was found in tap water in Korea. This study investigated whether Vermamoeba induced allergic airway inflammation in mice. We selected 2 free-living amoebas (FLAs) isolated from tap water, which included Korean FLA 5 (KFA5; Vermamoeba vermiformis) and 21 (an homolog of Acanthamoeba lugdunensis KA/ E2). We axenically cultured KFA5 and KFA21. We applied approximately 1 × 106 to mice’s nasal passages 6 times and investigated their pathogenicity. The airway resistance value was significantly increased after KFA5 and KFA21 treatments. The eosinophil recruitment and goblet cell hyperplasia were concomitantly observed in bronchial alveolar lavage (BAL) fluid and lung tissue in mice infected with KFA5 and KFA21. These infections also activated the Th2-related interleukin 25, thymic stromal lymphopoietin, and thymus and activation-regulated chemokines gene expression in mouse lung epithelial cells. The CD4+ interleukin 4+ cell population was increased in the lung, and the secretion of Th2-, Th17-, and Th1-associated cytokines were upregulated during KFA5 and KFA21 infection in the spleen, lung-draining lymph nodes, and BAL fluid. The pathogenicity (allergenicity) of KFA5 and KFA21 might not have drastically changed during the long-term in vitro culture. Our results suggested that Vermamoeba could elicit allergic airway inflammation and may be an airway allergen.

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Section: Discussionmentioning

confidence: 99%

Free-Living Amoeba Vermamoeba vermiformis Induces Allergic Airway Inflammation

et al. 2022

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“…Beside the classification according to cyst size and shape (group I–III) (Marciano-Cabral and Cabral 2003 ), Acanthamoeba spp. based on the SSU rDNA can be divided into 23 genotypes (T1–T23) (Norouzi et al 2021 ); however, most of the AK cases are caused by the T4 genotype (Jercic et al 2019 ). In this study, an Acanthamoeba sp.…”

Section: Resultsmentioning

confidence: 99%

Molecular evidence of Monocercomonas and Acanthamoeba in the feces of captive reptiles

et al. 2022

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Reptiles are frequently kept as pet animals. They are considered as important reservoirs of protozoa with veterinary-medical significance. At a reptile farm in Ireland, fecal samples were collected from 98 captive reptiles, representing 43 species of three orders (Squamata, Testudines, and Crocodylia). After DNA extraction, all samples were screened by conventional PCRs, targeting the ribosomal small subunit (SSU) RNA and alpha-tubulin genes of trichomonads and SSU RNA gene of Acanthamoeba spp. One leopard gecko (Eublepharis macularius) was positive for a not yet reported species/genotype of the genus Monocercomonas, different from M. colubrorum. Various Acanthamoeba genotypes were detected in six reptilian species, i.e., Acanthamoeba genotype T11 in Eunectes notaeus and Heloderma suspectum/horridum; genotype T4 in Varanus exanthematicus, Chlamydosaurus kingii, and Macrochelys temminckii; and the genotype T13 in Iguana iguana. Some of these amoeba species might have clinicopathological significance in both humans and animals. Our findings highlight the importance to monitor pathogenic protozoa in pet as well as wildlife reptiles, as a source of possible infection for animals and humans living nearby.

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“…The ameba trophozoites remained in an irregular shape because of their undisrupted acanthopodia, flattened onto the contact lens, and coated with bacteria. Biofilms produced by microorganisms have been shown to help protect ameba [ 39 ]. The biofilms produced by S. aureus and P. aeruginosa may protect Acanthamoeba from the cytotoxic effect of P. betle leaf extract, which requires further investigation.…”

Section: Discussionmentioning

confidence: 99%

Inhibitory and anti-adherent effects of Piper betle L. leaf extract against Acanthamoeba triangularis in co-infection with Staphylococcus aureus and Pseudomonas aeruginosa: A sustainable one-health approach

Kwankaew,

Sangkanu,

Mitsuwan

et al. 2024

Vet World

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Background and Aim: Keratitis is a serious ocular infection often caused by pathogenic micro-organisms such as Acanthamoeba spp. Among other harmful microbes, Acanthamoeba keratitis presents a particular challenge due to its resistance to conventional antimicrobial agents. Piper betle Linn., commonly known as betel leaf, has been traditionally used for its medicinal properties. This study aimed to assess the potential of the leaf ethanol extract of P. betle Linn. in the treatment of Acanthamoeba triangularis in monoculture and co-culture with two prevalent pathogenic bacteria, Staphylococcus aureus and Pseudomonas aeruginosa, associated with keratitis. Materials and Methods: Minimum inhibitory concentrations (MICs) of A. triangularis, S. aureus, and P. aeruginosa extracts in monoculture and coinfected conditions were examined. In addition, this study explored the potential of the extract in preventing Acanthamoeba adherence in both monoculture and co-culture environments. Scanning electron microscopy (SEM) analysis confirmed the impact of the extract on Acanthamoeba cell membranes, including acanthopodia. Furthermore, a time-kill kinetic assay was used to validate the amoebicidal activity of the extract against A. triangularis and the tested bacteria. Results: MICs for trophozoites, cysts, P. aeruginosa, and S. aureus in the monoculture were 0.25, 0.25, 0.51, and 0.128 mg/mL, respectively, whereas the MICs for Acanthamoeba coinfected with bacteria were higher than those in the monoculture. This extract inhibited the growth of A. triangularis trophozoites and cysts for up to 72 h. Moreover, P. betle extract effectively prevented the adherence of Acanthamoeba to contact lenses under monoculture conditions. SEM analysis confirmed that P. betle extract affects the cell membrane of Acanthamoeba, including Acanthopodia. In addition, the time-kill kinetic assay confirmed that the extract contained amoebicidal activity against A. triangularis, including the tested bacteria. Notably, S. aureus was more susceptible than A. triangularis and P. aeruginosa to P. betle extract treatment. Unexpectedly, our study revealed that S. aureus negatively affected A. triangularis in the co-culture after 3 days of incubation, whereas P. aeruginosa facilitated the growth of A. triangularis in the presence of the extract. Conclusion: This study provides compelling evidence of the anti-adhesive and anti-Acanthamoeba properties of P. betle leaf extract against A. triangularis under monoculture and co-culture conditions. The observed impact on Acanthamoeba cell membranes, coupled with the time-kill kinetic assay results, underscores the potential of P. betle leaf extract as a promising agent for combating Acanthamoeba-related infections in humans and animals. Keywords: Piper betle extract, Acanthamoeba triangularis, co-infection, keratitis, Pseudomonas aeruginosa, Staphylococcus aureus.

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Molecular Identification of Pathogenic Free-Living Amoeba from Household Biofilm Samples in Iran: A Risk Factor for Acanthamoeba Keratitis

Cited by 10 publications

References 41 publications

Free-Living Amoeba Vermamoeba vermiformis Induces Allergic Airway Inflammation

Free-Living Amoeba Vermamoeba vermiformis Induces Allergic Airway Inflammation

Molecular evidence of Monocercomonas and Acanthamoeba in the feces of captive reptiles

Inhibitory and anti-adherent effects of Piper betle L. leaf extract against Acanthamoeba triangularis in co-infection with Staphylococcus aureus and Pseudomonas aeruginosa: A sustainable one-health approach

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