Molecular detection and comparison of Acanthamoeba genotypes in different functions of watersheds in Taiwan

Kao, Po Min; Hsu, Bing-Mu; Chen, Nai Hsiung; Huang, Kuan Hao; Huang, Chin Chun; Ji, Dar Der; Chen, Jung‐Sheng; Lin, Wei Chen; Huang, Shih Wei; Chiu, Yi Chou

doi:10.1007/s10661-011-2267-4

Cited by 16 publications

(11 citation statements)

References 39 publications

(29 reference statements)

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“…However, many species of Acanthamoeba are potentially pathogenic for animals and for humans. A. castellanii, A. polyphaga , and A. culbertsoni are the most common species to infect humans [30, 31]. Acanthamoeba spp.…”

Section: Discussionmentioning

confidence: 99%

Isolation and Identification of Free-Living Amoebae from Tap Water in Sivas, Turkey

Coşkun

Özçelık

Tutar

et al. 2013

BioMed Research International

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The present work focuses on a local survey of free-living amoebae (FLA) that cause opportunistic and nonopportunistic infections in humans. Determining the prevalence of FLA in water sources can shine a light on the need to prevent FLA related illnesses. A total of 150 samples of tap water were collected from six districts of Sivas province. The samples were filtered and seeded on nonnutrient agar containing Escherichia coli spread. Thirty-three (22%) out of 150 samples were found to be positive for FLA. The FLA were identified by morphology and by PCR using 18S rDNA gene. The morphological analysis and partial sequencing of the 18S rDNA gene revealed the presence of three different species, Acanthamoeba castellanii, Acanthamoeba polyphaga, and Hartmannella vermiformis. Naegleria fowleri, Balamuthia mandrillaris, or Sappinia sp. was not isolated during the study. All A. castellanii and A. polyphaga sequence types were found to be genotype T4 that contains most of the pathogenic Acanthamoeba strains. The results indicated the occurrence and distribution of FLA species in tap water in these localities of Sivas, Turkey. Furthermore, the presence of temperature tolerant Acanthamoeba genotype T4 in tap water in the region must be taken into account for health risks.

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Section: Discussionmentioning

confidence: 99%

Isolation and Identification of Free-Living Amoebae from Tap Water in Sivas, Turkey

Coşkun

Özçelık

Tutar

et al. 2013

BioMed Research International

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“…Furthermore, the M3 method was shown the best sensitivity (96%) and accuracy (97%) than other methods, and M1 method also was shown poor sensitivity (32 to 50%) and accuracy (41 to 56%) in whichever pretreatment process. These findings indicate that many studies involving the detection of environmental Acanthamoeba underestimated the detection rate by using single-step Genotyping (JDP) PCR 6 , 16 , 34 , 39 , 44 , 45 . Many environmental microbiology studies have proven the sensitivity of nested PCR and real-time PCR for the determination of the actual detection rate in the environment 29 , 30 .…”

Section: Resultsmentioning

confidence: 96%

“…Sensitivity and detection rates of culture-dependent procedure and direct molecular enumeration procedure for detecting Acanthamoeba in the aquatic environment are contradictory. Surprisingly, few reports have shown that the culture procedure is more sensitive than the direct concentration procedure while others have opposed the same 39 , 44 – 46 . Our results provide a reasonable explanation to support that the direct concentration procedure is usually sensitive than the culture-dependent procedure explaining the advantages and disadvantages of both methodologies.…”

Section: Resultsmentioning

confidence: 99%

Efficient nested-PCR-based method development for detection and genotype identification of Acanthamoeba from a small volume of aquatic environmental sample

Hsu

Chen

Tsai

et al. 2021

Sci Rep

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Acanthamoeba spp. are opportunistic human pathogens that cause granulomatous amoebic encephalitis and keratitis, and their accurate detection and enumeration in environmental samples is a challenge. In addition, information regarding the genotyping of Acanthamoeba spp. using various PCR methods is equally critical. Therefore, considering the diverse niches of habitats, it is necessary to develop an even more efficient genotyping method for Acanthamoeba spp. detection. This study improved the sensitivity of detection to avoid underestimation of Acanthamoeba spp. occurrence in aquatic environmental samples, and to accurately define the pathogenic risk by developing an efficient PCR method. In this study, a new nested genotyping method was established and compared with various PCR-based methods using in silico, lab, and empirical tests. The in silico test showed that many PCR-based methods could not successfully align specific genotypes of Acanthamoeba, except for the newly designed nested PCR and real-time PCR method. Furthermore, 52 water samples from rivers, reservoirs, and a river basin in Taiwan were analysed by six different PCR methods and compared for genotyping and detection efficiency of Acanthamoeba. The newly developed nested-PCR-based method of genotyping was found to be significantly sensitive as it could effectively detect the occurrence of Acanthamoeba spp., which was underestimated by the JDP-PCR method. Additionally, the present results are consistent with previous studies indicating that the high prevalence of Acanthamoeba in the aquatic environment of Taiwan is attributed to the commonly found T4 genotype. Ultimately, we report the development of a small volume procedure, which is a combination of recent genotyping PCR and conventional real-time PCR for enumeration of aquatic Acanthamoeba and acquirement of biologically meaningful genotyping information. We anticipate that the newly developed detection method will contribute to the precise estimation, evaluation, and reduction of the contamination risk of pathogenic Acanthamoeba spp., which is regularly found in the water resources utilised for domestic purposes.

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“…The detection of Acanthamoeba in the temperate region, subtropical region, and tropical region was in the range of 7–94% [6–13], 3.6–21.2% [14–19], and 13.2–43.2% [20–26], respectively. In these regions, the water temperature differs according to the rivershed.…”

Section: Introductionmentioning

confidence: 99%

Diversity and Seasonal Impact ofAcanthamoebaSpecies in a Subtropical Rivershed

Kao

Chou

Tao

et al. 2013

BioMed Research International

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This study evaluated the presence of Acanthamoeba species in the Puzih River watershed, which features typical subtropical monsoon climate and is located just above the Tropic of Cancer in Taiwan. The relationship between the seasonal and geographical distributions of Acanthamoeba species in this rivershed was also investigated. Acanthamoeba species were detected in water samples using the amoebal enrichment culture method and confirmed by PCR. A total of 136 water samples were included in this study, 16 (11.7%) of which contained Acanthamoeba species. Samples with the highest percentage of Acanthamoeba (32.4%) were obtained during the summer season, mainly from upstream areas. The identified species in the four seasons included Acanthamoeba palestinensis (T2), Acanthamoeba sp. IS2/T4 (T4), Acanthamoeba lenticulata (T5), Acanthamoeba hatchetti (T11), Acanthamoeba healyi (T12), and Acanthamoeba jacobsi (T15). The most frequently identified Acanthamoeba genotype was T4 (68.7%). Acanthamoeba genotype T4 is responsible for Acanthamoeba keratitis and should be considered for associated human health risk potential in the rivershed.

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Molecular detection and comparison of Acanthamoeba genotypes in different functions of watersheds in Taiwan

Cited by 16 publications

References 39 publications

Isolation and Identification of Free-Living Amoebae from Tap Water in Sivas, Turkey

Isolation and Identification of Free-Living Amoebae from Tap Water in Sivas, Turkey

Efficient nested-PCR-based method development for detection and genotype identification of Acanthamoeba from a small volume of aquatic environmental sample

Diversity and Seasonal Impact ofAcanthamoebaSpecies in a Subtropical Rivershed

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