Droplet digital PCR as a tool for investigating dynamics of cryptic symbionts

Hiillos, Anna-Lotta; Thonig, Anne; Knott, K. Emily

doi:10.1002/ece3.8372

Cited by 3 publications

(15 citation statements)

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“…Facilitation could occur, for example, by suppression of the host immune system, as has been suspected of parasitic mites and wing deforming virus in honeybees (Nazzi et al, 2012) and myxoma virus and the nematode Trichostrongylus retortaeformis infections in rabbits (Cattadori et al, 2007). Coinfecting parasites could, alternatively, antagonize each other within the host by competing for space or energetic resources (Clerc et The apicomplexans studied here infect the intestine of P. elegans (Hiillos et al, 2021;Paskerova et al, 2018), a circumstance that could lead to a negative outcome through competition of host resources (Dallas et al, 2019). However, as dual infections were relatively frequent, and Rhytidocystis sp.…”

Section: Discussionmentioning

confidence: 98%

“…It has an important role in benthic communities, as it can reach high densities (Bolam, 2004 ; Bolam & Fernandes, 2003 ) and is an important prey animal to other invertebrates and fish (Mattila, 1997 ). At least three protist species in the phylum Apicomplexa are known to infect P. elegans : an archigregarine, Selenidium pygospionis (Paskerova et al, 2018 ); an eugregarine Polyrhabdina pygospionis (Paskerova et al, 2021 ); and an undescribed marosporidian (Class Marosporida; Mathur et al, 2020 ) in the genus Rhytidocystis (Hiillos et al, 2021 ), from now on referred to as Rhytidocystis sp. All of these apicomplexans infect the worm's intestine.…”

Section: Methodsmentioning

confidence: 99%

“…Host samples were collected from three populations, Cramond Beach (55°58′N, 3°17′53 W, Edinburgh, UK), Herslev (55°40′N, 11°59′E, DK), and Vellerup (55°44′N 11°52′E, DK). These populations were chosen because they have been studied extensively previously (Bolam, 2004 ; Hiillos et al, 2021 ; Thonig et al, 2016 , 2017 ). All samples were collected in early November in consecutive years 2018 and 2019.…”

Section: Methodsmentioning

confidence: 99%

“…The cox1 gene is also currently the only available genetic marker for Rhytidocystis sp. (Hiillos et al, 2021). Additionally, because another P. elegans-infecting gregarine (Polyrhabdina pygospionis) was not studied here, the use of cox1 ensured the assays were specific, as P. pygospionis has lost cox1 from its mitochondrial genome (Mathur et al, 2021;Salomäki et al, 2021).…”

Section: Apicomplexan Detection and Quantificationmentioning

confidence: 99%

“…After amplification, the fraction of positive droplets is determined and the concentration of the target can be estimated (Hindson et al, 2011(Hindson et al, , 2013. As ddPCR gives absolute quantification and no standard curves are needed, it is particularly useful in quantifying symbionts that have little genetic information available and are unculturable outside their hosts (Hiillos et al, 2021).…”

Section: Apicomplexan Detection and Quantificationmentioning

confidence: 99%

See 4 more Smart Citations

Coinfection patterns of two marine apicomplexans are not associated with genetic diversity of their polychaete host

Hiillos

Rony

Rueckert

et al. 2022

J Eukaryotic Microbiology

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Coinfections of two or more parasites within one host are more of a rule than an exception in nature. Interactions between coinfecting parasites can greatly affect their abundance and prevalence. Characteristics of the host, such as genetic diversity, can also affect the infection dynamics of coinfecting parasites.Here, we investigate for the first time the association of coinfection patterns of two marine apicomplexans, Rhytidocystis sp. and Selenidium pygospionis, with the genetic diversity of their host, the polychaete Pygospio elegans, from natural populations. Host genetic diversity was determined with seven microsatellite loci and summarized as allelic richness, inbreeding coefficient, and individual heterozygosity. We detected nonsignificant correlations between infection loads and both individual host heterozygosity and population genetic diversity.Prevalence and infection load of Rhytidocystis sp. were higher than those of S. pygospionis, and both varied spatially. Coinfections were common, and almost all hosts infected by S. pygospionis were also infected by Rhytidocystis sp. Rhytidocystis sp. infection load was significantly higher in dual infections.Our results suggest that factors other than host genetic diversity might be more important in marine apicomplexan infection patterns and experimental approaches would be needed to further determine how interactions between the apicomplexans and their host influence infection.

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Section: Discussionmentioning

confidence: 98%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Apicomplexan Detection and Quantificationmentioning

confidence: 99%

Section: Apicomplexan Detection and Quantificationmentioning

confidence: 99%

See 3 more Smart Citations

Coinfection patterns of two marine apicomplexans are not associated with genetic diversity of their polychaete host

Hiillos

Rony

Rueckert

et al. 2022

J Eukaryotic Microbiology

Self Cite

View full text Add to dashboard Cite

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Using digital PCR to predict ciliate abundance from ribosomal RNA gene copy numbers

Gross,

Dunthorn,

Mauvisseau

et al. 2024

Environmental Microbiology

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Ciliates play a key role in most ecosystems. Their abundance in natural samples is crucial for answering many ecological questions. Traditional methods of quantifying individual species, which rely on microscopy, are often labour‐intensive, time‐consuming and can be highly biassed. As a result, we investigated the potential of digital polymerase chain reaction (dPCR) for quantifying ciliates. A significant challenge in this process is the high variation in the copy number of the taxonomic marker gene (ribosomal RNA [rRNA]). We first quantified the rRNA gene copy numbers (GCN) of the model ciliate, Paramecium tetraurelia, during different stages of the cell cycle and growth phases. The per‐cell rRNA GCN varied between approximately 11,000 and 130,000, averaging around 50,000 copies per cell. Despite these variations in per‐cell rRNA GCN, we found a highly significant correlation between GCN and cell numbers. This is likely due to the coexistence of different cellular stages in an uncontrolled (environmental) ciliate population. Thanks to the high sensitivity of dPCR, we were able to detect the target gene in a sample that contained only a single cell. The dPCR approach presented here is a valuable addition to the molecular toolbox in protistan ecology. It may guide future studies in quantifying and monitoring the abundance of targeted (even rare) ciliates in natural samples.

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Heterogeneous distribution of Cardinium in whitefly populations is associated with host nuclear genetic background

Liu

Wei

et al. 2023

Insect Science

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Inherited bacterial symbionts are very common in arthropods, but infection frequency can vary widely among populations. Experiments and interpopulation comparisons suggest that host genetic background might be important in explaining this variation. Our extensive field investigation showed that the infection pattern of the facultative symbiont Cardinium was heterogeneous across geographical populations of the invasive whitefly Bemisia tabaci Mediterranean (MED) in China, with genetic nuclear differences evident in 2 of the populations: 1 with a low infection rate (SD line) and 1 with a high infection rate (HaN line). However, whether the heterogeneous frequency of Cardinium is associated with the host genetic background remains poorly understood. Here, we compared the fitness of the Cardinium‐infected and uninfected sublines with similar nuclear genetic backgrounds from SD and HaN lines, respectively, and further determine whether host extranuclear or nuclear genotype influenced the Cardinium–host phenotype by performing 2 new introgression series of 6 generations between SD and HaN lines (i.e., Cardinium‐infected females of SD were backcrossed with uninfected males of HaN, and vice versa). The results showed that Cardinium provides marginal fitness benefits in the SD line, whereas Cardinium provides strong fitness benefits in the HaN line. Further, both Cardinium and the Cardinium–host nuclear interaction influence the fecundity and pre‐adult survival rate of B. tabaci, whereas the extranuclear genotype does not. In conclusion, our results provide evidence that Cardinium‐mediated fitness effects were closely associated with the host genetic background, which provides a fundamental basis for understanding the mechanism underlying the heterogeneous distribution of Cardinium in B. tabaci MED populations across China.

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Droplet digital PCR as a tool for investigating dynamics of cryptic symbionts

Cited by 3 publications

References 84 publications

Coinfection patterns of two marine apicomplexans are not associated with genetic diversity of their polychaete host

Coinfection patterns of two marine apicomplexans are not associated with genetic diversity of their polychaete host

Using digital PCR to predict ciliate abundance from ribosomal RNA gene copy numbers

Heterogeneous distribution of Cardinium in whitefly populations is associated with host nuclear genetic background

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