Update on Acanthamoeba jacobsi genotype T15, including full-length 18S rDNA molecular phylogeny

Corsaro, Daniele; Köhsler, Martina; Filippo, Margherita Montalbano Di; Venditti, Danielle; Monno, Rosa; Cave, David Di; Berrilli, Federica; Walochnik, Julia

doi:10.1007/s00436-017-5406-1

Cited by 57 publications

(29 citation statements)

References 54 publications

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“…These stretches did not include low complexity spacers, which are difficult to align. Third, some of the strains showed greater differences from the Neff strain in each CWP, consistent with previous descriptions of Acanthamoeba strain diversity based upon 18S rDNA sequences (53). Fourth, while coding sequences and 5’ UTRs were well conserved, introns were very poorly conserved, with the exception of branch-point sequences.…”

Section: Resultssupporting

confidence: 88%

The most abundant cyst wall proteins ofAcanthamoeba castellaniiare cellulose-binding lectins from three gene families that localize to distinct structures in cyst walls

Samuelson

Magistrado-Coxen

Aqeel

et al. 2018

Preprint

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18Acanthamoeba castellanii, cause of keratitis and blindness, is an emerging pathogen because 19 of its association with contact lens use. The cyst wall contributes to pathogenesis as cysts are 20 resistant to sterilizing reagents in lens solutions and to antibiotics applied to the eye. We used 21 transmission electron microscopy, as well as structured illumination microscopy and probes for 22 cellulose and chitin, to show that purified cyst walls of A. castellanii retain an outer ectocyst 23 layer, an inner endocyst layer, and conical ostioles that connect the layers. Mass spectrometry 24 showed candidate cyst wall proteins are dominated by three families of lectins (named here 25 Luke, Leo, and Jonah), each of which binds to microcrystalline cellulose and to a lesser degree 26 chitin. A Jonah lectin, which has one choice-of-anchor A (CAA) domain, localizes to the 27 ectocyst layer of mature cyst walls. Luke lectins, which have two or three carbohydrate-binding 28 modules (CBM49), localize to the endocyst layer and ostioles. A Leo lectin, which has two 29 domains with eight Cys residues each (8-Cys), also localizes to the endocyst layer and 30 ostioles. In summary, the most abundant A. castellanii cyst wall proteins are three sets of 31 lectins, which have carbohydrate-binding modules that are conserved (CBM49s of Luke), 32 newly characterized (CAA of Jonah), or unique to Acanthamoebae (8-Cys of Leo). Despite 33 their lack of common ancestry, Luke and Leo lectins both localize to the endocyst layer and 34 ostioles, while the Jonah lectin localizes to the ectocyst layer. 35IMPORTANCE 36 Fifty years ago, investigators identified cellulose in the Acanthamoeba cyst wall, which has an 37 outer ectocyst layer, an inner endocyst layer, and conical ostioles that connect the layers. 38Here we show cyst walls also contain chitin and three large sets of cellulose-and chitin-binding 39 lectins, which have distinct localizations. The Acanthamoeba cyst wall, therefore, is more 40 complicated than cyst walls of Entamoeba and Giardia (causes of dysentery and diarrhea, 41 respectively), which have a single layer, a single glycopolymer, and small sets of one or two 42 lectins. In contrast, the Acanthamoeba cyst wall is far simpler than the walls of fungi and 43 plants, which have multiple layers, numerous glycopolymers, and hundreds of proteins. In 44 addition to providing a better understanding of the cell biology and biochemistry of the 45 Acanthamoeba cyst wall, these studies may lead to diagnostic antibodies that bind to cyst wall 46 proteins and/or therapeutics that target chitin. 47 100 file S1) (41). The GST-AcCBM49 expression construct was designed to replicate that used to 101 determine the carbohydrate binding properties of SlCBM49, which is a C-terminal 102 carbohydrate-binding module of the Solanum lycopersicum (tomato) cellulase SlGH9C (42). In 103 both intact cysts and purified cyst walls, GST-AcCBM49 predominantly labeled the ectocyst 104 layer, WGA highlighted the ostioles, and CFW stained the endocyst...

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Section: Resultssupporting

confidence: 88%

The most abundant cyst wall proteins ofAcanthamoeba castellaniiare cellulose-binding lectins from three gene families that localize to distinct structures in cyst walls

Samuelson

Magistrado-Coxen

Aqeel

et al. 2018

Preprint

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“…that could be grown at 37 • C or even at 44 • C (such as A. jacobsi). The identified thermophilic Acanthamoeba included A. lenticulata [36], A. castellanii [37], A. byersi [38], A jacobsi [39] and A. hatchetti [40], all known to be involved in pathogenic processes. Unfortunately, our methodology did not allow us to identify the different Acanthamoeba genotypes [41].…”

Section: Discussionmentioning

confidence: 99%

Cartography of Free-Living Amoebae in Soil in Guadeloupe (French West Indies) Using DNA Metabarcoding

et al. 2020

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Free-living amoebae (FLA) are ubiquitous protists. Pathogenic FLA such as N. fowleri can be found in hot springs in Guadeloupe, soil being the origin of this contamination. Herein, we analyzed the diversity and distribution of FLA in soil using a targeted metataxonomic analysis. Soil samples (n = 107) were collected from 40 sites. DNA was extracted directly from soil samples or from FLA cultivated at different temperatures (30, 37 and 44 °C). Metabarcoding studies were then conducted through FLA 18SrDNA amplicons sequencing; amplicon sequence variants (ASV) were extracted from each sample and taxonomy assigned against SILVA database using QIIME2 and SHAMAN pipelines. Vermamoeba were detected in DNA extracted directly from the soil, but to detect other FLA an amoebal enrichment step was necessary. V. vermiformis was by far the most represented species of FLA, being detected throughout the islands. Although Naegleria were mainly found in Basse-Terre region, N. fowleri was also detected in Grand Terre and Les Saintes Islands. Acanthamoeba were mainly found in areas where temperature is approx. 30 °C. Vannella and Vahlkampfia were randomly found in Guadeloupe islands. FLA detected in Guadeloupe include both pathogenic genera and genera that can putatively harbor microbial pathogens, therefore posing a potential threat to human health.

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“…can complete their life cycle, environmental performance without having to enter the human or animal host [7,8]. The genus Acanthamoeba has been currently classified into 21 different genotypes, T1-T21, based on 18S rRNA nucleotide sequence [9,10]. Some genera of Acanthamoeba cause different infections, which produce Acanthamoeba keratitis (AK), subacute or chronic granulomatous amoebic encephalitis, and skin infections.…”

Section: Introductionmentioning

confidence: 99%

Genotyping of Acantamoeba spp. from rhisophere in Hungary

Orosz

Posta

2020

AMicr

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The protista Acanthamoeba is a free-living amoeba existing in various environments. A number of species among protista are recognized as human pathogens, potentially causing Acanthamoeba keratitis (AK), granulomatous amoebic encephalitis (GAE), and chronic granulomatous lesions. In this study, 10 rhizosphere samples were collected from maize and alfalfa plants in experimental station at Institute of Genetics, Microbiology and Biotechnology, Szent István University. We detected Acanthamoeba based on the quantitative real-time PCR assay and sequence analysis of the 18S rRNA gene. All studied molecular biological methods are suitable for the detection of Acanthamoeba infection in humans. The quantitative real-time PCR-based methods are more sensitive, simple, and easy to perform; moreover, these are opening avenue to detect the effect of number of parasites on human disease. Acanthamoeba species were detected in five (5/10; 50%) samples. All Acanthamoeba strains belonged to T4 genotype, the main AK-related genotype worldwide. Our result confirmed Acanthamoeba strains in rhizosphere that should be considered as a potential health risk associated with human activities in the environment.

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Update on Acanthamoeba jacobsi genotype T15, including full-length 18S rDNA molecular phylogeny

Cited by 57 publications

References 54 publications

The most abundant cyst wall proteins ofAcanthamoeba castellaniiare cellulose-binding lectins from three gene families that localize to distinct structures in cyst walls

The most abundant cyst wall proteins ofAcanthamoeba castellaniiare cellulose-binding lectins from three gene families that localize to distinct structures in cyst walls

Cartography of Free-Living Amoebae in Soil in Guadeloupe (French West Indies) Using DNA Metabarcoding

Genotyping of Acantamoeba spp. from rhisophere in Hungary

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