Next generation amplicon sequencing improves detection of Blastocystis mixed subtype infections

Maloney, Jenny G.; Molokin, Aleksey; Santı́n, Mónica

doi:10.1016/j.meegid.2019.04.013

Cited by 69 publications

(63 citation statements)

References 25 publications

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“…The use of NGS could aid in understanding host‐parasite specific epidemiological cycling in nature because of its capacity to resolve a higher level of genetic diversity identifying low abundance of Blastocystis STs within the same host (Maloney et al. 2019b). ST7 was the most common ST found causing single or mixed infections in combination with other Blastocystis STs in three foxes and a common genet.…”

Section: Resultsmentioning

confidence: 99%

“…No mixed subtype infections were found using the traditional method, barcoding primers coupled with Sanger sequencing, possibly due to the inherent nature of PCR, which preferentially amplifies the predominant subtypes that will be identified by Sanger sequencing. The use of NGS could aid in understanding host-parasite specific epidemiological cycling in nature because of its capacity to resolve a higher level of genetic diversity identifying low abundance of Blastocystis STs within the same host (Maloney et al 2019b). ST7 was the most common ST found causing single or mixed infections in combination with other Blastocystis STs in three foxes and a common genet.…”

Section: Resultsmentioning

confidence: 99%

“…Next‐generation amplicon sequencing (NGS) libraries were prepared and analysed as previously described (Maloney et al. 2019b). Briefly, all samples were screened by PCR using primers ILMN_Blast505_532F and ILMN_Blast998_1017R.…”

Section: Methodsmentioning

confidence: 99%

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Blastocystis sp. Subtype Diversity in Wild Carnivore Species from Spain

Calero‐Bernal

Santı́n

Maloney

et al. 2019

J Eukaryotic Microbiology

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The occurrence and molecular diversity of the stramenopile eukaryote Blastocystis sp. was investigated by PCR and sequencing (Sanger and NGS) methods in 380 faecal specimens of free‐living carnivores in Spain. Blastocystis sp. was confirmed in 1.6% (6/380) of the specimens analysed. Two samples from a common genet and a fox were successfully subtyped as ST7 by Sanger. Using NGS, ST14 was found in a fox and a European polecat, ST7 in a fox, and two additional foxes presented mixed infections of ST1/ST2/ST4 and ST1/ST2/ST7, respectively. Wild carnivore species could act as carriers of zoonotic Blastocystis subtypes.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Blastocystis sp. Subtype Diversity in Wild Carnivore Species from Spain

Calero‐Bernal

Santı́n

Maloney

et al. 2019

J Eukaryotic Microbiology

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“…PCR ampli cation coupled with sequencing, preferentially ampli ed and allowed to identify the predominant subtypes. Nevertheless, as previously demonstrated, the cloning of the PCR product or next generation amplicon sequencing is required to discriminate different subtypes in the same host [3,24,25,26].…”

Section: Resultsmentioning

confidence: 99%

“…Blastocystis is a common intestinal protozoon distributed worldwide, infecting humans and a wide range of domestic and wild animals. Molecular studies based on sequence analysis of the small subunit (SSU) ribosomal RNA evidenced an extensive genetic diversity allowing the identi cation in mammalian and avian hosts of at least 26 divergent lineages, termed subtypes (STs), which could be considered as separate species [1,2,3]. Ten of the 26 subtypes, ST1 to ST9, and ST12 have been identi ed in human samples and have also been reported in animals so far [4,5,6].…”

Section: Introductionmentioning

confidence: 99%

Molecular detection of Blastocystis from animals in Italy: subtypes distribution and implications for the zoonotic transmission

Gabrielli

Furzi

Brianti

et al. 2020

Preprint

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Background: Blastocystis is a common intestinal protist distributed worldwide infecting humans and a wide range of domestic and wild animals. It exhibits an extensive genetic diversity and so far, 26 subtypes (STs) have been identified in animal hosts, ten of them (ST1-ST9 and ST12) reported in humans with varying prevalence. Since several STs are common to humans and animals it has been proposed that a proportion of human infections may have a zoonotic origin. Aims of the present study were to: 1) genetically detect Blastocystis in faecal samples of farmed animals and wild carnivores; 2) investigate the distribution of Blastocystis STs in different animal hosts; 3) provide a first study on the Blastocystis STs circulating between animals and humans in Italy.Methods: Fresh faecal samples (N=269) were collected from carnivores and farmed animals in different Italian provinces and submitted to genomic DNA extraction and PCR amplification followed by both sequence and phylogenetic analysis (Neighbour Joining and Maximum Parsimony)Results: Blastocystis was detected in 50% of the farmed animals (42 out of 84), and 19 of them were successfully subtyped. Conversely, all the faecal samples (N=185) from domestic and wild carnivores (dogs, cats, foxes) tested in the present study, resulted negative. Phylogenetic analysis showed the finding of ST5, ST7, ST9 and ST10 in the samples from animals. The comparison with sequences of Blastocystis STs previously detected from humans in Italy showed the ST7, as a potential source of zoonotic transmission.Conclusions: The present study represents the widest epidemiological survey so far performed in animals in Italy. Further epidemiological studies using molecular approaches are required to determine the occurrence and distribution of Blastocystis STs in other potential animal reservoirs and to define the pathways of zoonotic transmission.

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