Isolation of potentially pathogenic strains of Acanthamoeba in wild squirrels from the Canary Islands and Morocco

Lorenzo‐Morales, Jacob; López-Darias, Marta; Martínez-Carretero, Enrique; Valladares, Basilio

doi:10.1016/j.exppara.2007.03.014

Cited by 24 publications

(12 citation statements)

References 28 publications

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“…Nevertheless, the cyst form of Acanthamoeba is resistant to chemical reagents, temperature, and pH. For this reason, a diversity of reagents-including ammonia, lysis buffer, proteinase K, and chelex 100-as well as increased duration times for incubation have been utilized to extract DNA for molecular studies (Iovieno et al 2011;Lehmann et al 1998;Lorenzo-Morales et al 2007;Pasricha et al 2003;Zhang et al 2004).…”

Section: Introductionmentioning

confidence: 99%

A simple PCR condition for detection of a single cyst of Acanthamoeba species

2011

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Acanthamoeba is a free-living protozoan with a worldwide distribution in a variety of natural and artificial habitats. It has even been found in contact lens solution. Acanthamoeba spp. can cause infections such as granulomatous amoebic encephalitis and amoebic keratitis. Specific and sensitive diagnosis of Acanthamoeba infections can prevent clinical symptoms from worsening. Recently, PCR technique has been used for Acanthamoeba diagnosis. Unfortunately the dormant cyst of Acanthamoeba is resistant to chemical reagents; thus, most extraction of DNA uses a commercial DNA extraction kit for obtaining DNA for further use in polymerase chain reaction (PCR). Therefore in the present study, we improved the ability to diagnose Acanthamoeba using a simplified PCR technique. Interestingly, heating at 94°C for 10 min could release DNA which is amplified with specific primers designed from 16S rRNA. The PCR product is about 180 bp. This technique is a simple and efficient method for detection of Acanthamoeba-even a single cyst-and does not require high-cost reagents or complicated procedures to extract DNA.

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Section: Introductionmentioning

confidence: 99%

A simple PCR condition for detection of a single cyst of Acanthamoeba species

2011

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“…High percentage of Acanthamoeba spp. in water, is a hygienic risk for public health especially for individuals with immune deficiency situation and use in recreational water sources [26].…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Acanthamoeba Species from Tehran Surface Water Sources Using Real Time PCR Method

2014

International Conference on Food, Biological and Medical Sciences (FBMS-2014) Jan. 28-29, 2014 Bangkok (Thailand)

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Acanthamoeba spp. is the ubiquitous potentially pathogenic free-living amoebae in nature such as water sources. Recently Acanthamoeba keratitis (Ak) continue to rise in Iran. Most of patients report a history of contact with water sources before the onset of disease. The main aim of the present study was to determine the occurrence of Acanthamoeba spp. in the recreational water sources of Tehran, Iran using morphological and molecular based tests. A total of 70 surface water samples were collected from environmental sources, including parks pools and swimming pools. The samples were filtrated and transferred to non-nutrient agar plates seeded with Escherichia coli and incubated for 2 to 7 days at 30°C or 42°C. The plates were examined by microscopy to morphologically identify Acanthamoeba spp. Following DNA extraction, PCR was used to confirm the microscopically identification. A total of 11 out of 35 samples of parks pools and 3 out of 35 of swimming pools were positive for Acanthamoeba species based on the morphological criteria, and all were confirmed by PCR method. The significant frequency of Acanthamoeba spp in surface water sources of Tehran is of importance to beconsidered from public health point of view by authorities.

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“…was diagnosed in Dakar (Ndiaye et al, 2005). The extents to which free living amoebae are present in water sources associated with human activities have been previously studied in Egypt (Mansour et al, 1991;Hamadto et al, 1993;Sadaka et al, 1994;LorenzoMorales et al, 2006) and Morocco (Lorenzo-Morales et al, 2007). Morphological study enabled us to detect morphotypic diversity.…”

Section: Discussionmentioning

confidence: 99%

Free-living amoebae (FLA): detection, morphological and molecular identification ofAcanthamoebagenus in the hydraulic system of an haemodialysis unit in Tunisia

Dendana¹,

Sellami²,

Jarraya

et al. 2008

Parasite

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Summary :The free-living amoebae (FLA) are ubiquitous and opportunistic protozoa. They can induce human and animal diseases. The aim of our study was to detect the FLA and Acanthamoeba genus in the hydraulic system of an hemodialysis unit. It was a prospective study of 46 water samples. The first collect (23) was before cleaning and after the haemodialysis sessions and the second (23) after cleaning and before the hemodialysis sessions. Results: the morphological study enabled us to detect morphotypic diversity. The predominant morphotypes were the acanthopodial forms (29 %). At the entrance of hemodialysis unit there were acanthopodial (44 %) and monotactic (25 %) forms; at the outlet, acanthopodial and fan-shaped forms (25 % each). In addition, Acanthamoeba genus was present in 39 % (1 st collect) and 18 % (2 nd collect). The amplification of the FLA 18S rDNA gene was negative in only one sample localized in the last stage of water treatment unit (WTU). The amplification of the 18S rDNA (ASA.A1) Acanthamoeba gene was positive in15 samples. Conclusions: we noted that, in the hemodialysis unit, the purification techniques used in the WTU were effective, but there is a problem of water stagnation in the drain, which constitutes an appropriate condition for the biofilms formation. It is then necessary to use a filter with a low porosity (0.2 µm) at the entrance of the hemodialysis unit and if possible to change the drain. Résumé

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Isolation of potentially pathogenic strains of Acanthamoeba in wild squirrels from the Canary Islands and Morocco

Cited by 24 publications

References 28 publications

A simple PCR condition for detection of a single cyst of Acanthamoeba species

A simple PCR condition for detection of a single cyst of Acanthamoeba species

Evaluation of Acanthamoeba Species from Tehran Surface Water Sources Using Real Time PCR Method

Free-living amoebae (FLA): detection, morphological and molecular identification ofAcanthamoebagenus in the hydraulic system of an haemodialysis unit in Tunisia

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