Monday, July 28, 2014

This Month In Blastocystis Research (JUL 2014)

For Spanish-speaking Blastocystis geeks, this summer must have been a real treat: Londoño-Franco and colleagues published a paper in Biomédica on Blastocystis in children and Colombia. But not only did they look for Blastocystis in faecal samples, they also sampled from finger nails, house floors, toys, tap water,  vegetables, other food items, etc... It is extremely rare to see studies aiming to identify sources of potential transmission, and I thought that this study would merit a blog post (unfortunately, I will have to rely on the Google translated version with all its potential limitations; I excuse for any misunderstandings).

Of course one of the big questions still remaining in Blastocystis research is: From where do we get this parasite? With more than one billion people colonised on the globe, the transmission pressure must be massive, and it's tempting to expect infectious cysts (or other stages) being more or less ubiquitous. There is some evidence accumulating that the parasite can be water-borne, and we also know that zoonotic transmission can occur (although relatively rarely, supposedly). However, this study takes things way further:

The authors carried out their study in Calarcá where they identified a prevalence of Blastocystis (based on microscopy of stool concentrates) of 57.5% in 275 children less than 5 years old; children aged 48 months or more were more prone to be positive than those who were younger. This is something we see a lot, and it either suggests a cumulative effect of colonisation (once established, colonisation is chronic), or that the behaviour (~exposure) or intestinal microbiota of older children favours colonisation.
Agua de panela (source).

Blastocystis was also found in dogs (63.3%), cats (56.3%), and poultry (35.7%). Moreover, it was found in tap water (38.5%), on toys (29.9%), baby bottles (18.5%), and under the nails of infected children (42.2%), their siblings (44.8%), and their mothers (34.2%). Among the vegetables that are typically consumed raw, it was found most frequently in lettuce (66.7%), and, in descending order, in tomato (44.4%), carrots (37.5%), cabbage (28.6%) and onion (25%). A high occurrence was seen in containers used to store 'aqua de panela', which is allegedly some kind of sugar water (haven't had the opportunity to sample it myself), with 47.7% of the samples positive. I believe that this drink is used as a sweetener and possibly also as a refreshment/energy drink, and maybe served with for instance cheese (image). Taken into account that Blastocystis is not exactly fussy about growth medium requirements, it may not be surprising at all to learn that this type of drink serves as a perfect stronghold for Blastocystis

The authors also explored a number of other things, among them i) the relative occurrence of cysts and vacuolar stages in the different types of samples and ii) whether any symptoms experienced over the past month could be attributed to Blastocystis, and iii) risk factors for colonisation. However, Google translate plays tricks on me on some of these bits, so I won't try to go more into detail with these findings. Suffice to say that the approach of distinguishing between different stages should help researchers find out more about which stage(s) that is/are responsible for transmission. Also, if for instance vacuolar stages are found in agua de panela and not cysts, then this might indicate that Blastocystis is actually growing in the drink? Which again is interesting because this would mean that Blastocystis capable of infecting humans can grow at temperatures lower than 37 degrees C.

Now, I could only have great confidence in the diagnostic work carried out by this team; however, I would have absolutely loved molecular confirmation of all of these findings. Also, maybe it would have been an idea to try and culture some of the Blastocystis found on fomites and in food/water to test for viability, or, as mentioned by the authors themselves, to test for viability using trypan blue. However, the authors should be praised for their perseverance and ingenuity, and I hope that this study will inspire other colleagues to pursue and expand on these initiatives and ideas.

This month saw a number of different Blastocystis-related papers, among them a paper from Klimes et al. on issues with Blastocystis genome annotation and polyadenylation-mediated termination codon creation in nuclear mRNA transcripts. Moreover, there's a paper on population structure analysis of seven eukaryotic microbial lineages, including Blastocystis, that apparently makes it possible to infer variable impacts of genetic exchange in populations of predominantly clonal micro-pathogens  (in fact the authors used our MLST data for ST3 in their analyses!). Finally, our colleagues in České Budějovice have produced an interesting review on self-infections with parasites; in the paper they point to the traditional focus on sussing out the pathogenic potential of parasites instead of trying to identify the potentially positive effects of parasite colonisation. Definitely worth a read!


Londoño-Franco AL, Loaiza-Herrera J, Lora-Suárez FM, & Gómez-Marín JE (2014). [Blastocystis sp. frequency and sources among children from 0 to 5 years of age attending public day care centers in Calarcá, Colombia]. Biomedica : Revista del Instituto Nacional de Salud, 34 (2), 218-27 PMID: 24967927 

Klimeš V, Gentekaki E, Roger AJ, & Eliáš M (2014). A large number of nuclear genes in the human parasite Blastocystis require mRNA polyadenylation to create functional termination codons. Genome Biology and Evolution PMID: 25015079 

Lukeš J, Kuchta R, Scholz T, & Pomajbíková K (2014). (Self-) infections with parasites: re-interpretations for the present. Trends in Parasitology PMID: 25033775

Tomasini N, Lauthier JJ, Ayala FJ, Tibayrenc M, & Diosque P (2014). How often do they have sex? A comparative analysis of the population structure of seven eukaryotic microbial pathogens. PLoS One, 9 (7) PMID: 25054834 

Thursday, July 3, 2014

This Month in Blastocystis Research (JUN 2014) - IMECs Edition

In June there was a paper out in Frontiers in Microbiology by Laura W Parfrey and co-workers identifying the diversity of intestinal microbial eukaryotic communities (IMECs) in humans and other mammals. It's probably one of the most interesting papers I've read for a long time; maybe because it expands on many of the things I've been blogging about - or at least intended to blog about (!) - over the past two years.

What the team did was to do comprehensive analysis of IMECs in both humans and mammals using broad specificity primers for PCR and next generation sequencing technology-based sequencing of the PCR products. While I'm not in a position to validate the analysis of the data, I'd just want to highlight the importance of the approach. It is very rare to see this type of analysis, despite the fact that it's probably the best currently available approach to studying the ecology, homeostasis and public health significance of IMECs. Some of these euks have probably co-evolved with humans and other animals over thousands and thousands of years and therefore may constitute part of the habitual/commensal flora; and so a current working hypothesis (Hygiene Theory) is that losing IMECs ('defaunation' due to Western life style (excessive hygiene and changes in diet)) may prove detrimental to human health and may be one of the most important reasons why we develop for instance allergies and other autoimmune diseases.

Blastocystis virtually obligate finding in Malawi citizens?
And indeed, what the authors found was that among 23 study individuals residing in agrarian communities in Malawi, Blastocystis and Entamoeba were almost obligate findings (not found in two infants, but apart from that almost a consistent finding), while none of the 13 (somewhat age-matched) study individuals from Boulder, Colorado, were infected with Blastocystis, and only two individuals had Entamoeba coli. I was surprised to read that Dientamoeba was not detected in any of the populations; it appears that there is a strong geographical component to the distribution of this parasite, but as the authors mention, specific tools are needed to confirm the absence.

The funny thing is that although this is not a paper specifically on Blastocystis, it is probably the most interesting surveys on Blastocystis coming from the US and a very valuable Blastocystis. Data on Blastocystis in this country is really scarce, but if the prevalence of the parasite is really as low as indicated in this study, then it's maybe quite understandable! And maybe (and this is a highly presumptuous 'maybe', I know) Blastocystis might even therefore an emerging pathogen in the US? When was the US experiencing the great IMECs wipe out? Can it be confirmed? Is there - within the US - also a strong geographical compoenent to the prevalence of IMECs?

Anyway, there are many interesting observations in the paper - and please visit the supporting files. Blastocystis ST11 was confirmed in an elephant (which also hosted Entamoeba moshkovskii! Probably first report of this parasite in an animal). ST13 was found in a Gazelle; not surprisingly, but nice to see independent data confirming what few researchers have found until now. ST4 was found in a sheep and in Okapis; when it comes to ST4, I'm hardly surprised about anything; it appears to be such a sporadic finding in a diversity of non-human hosts (i.e. low host specificity and incidental); one sheep also had ST8, a subtype almost exclusively seen in non-human primates (even South American monkeys rather than for instance African monkeys and apes), so this was surprising too. ST8 was moreover found in two kangaroos (not the first time), in an okapi (different from two first ones) which also hosted ST12, and in an armadillo!

Take home messages include:

1) The study is one of the first to virtually survey IMECs in human and non-human faecal samples using NGS tools.
2) The study confirms a very high prevalence of Blastocystis in some sub-Saharan African communities (for more on this, see a previous blog post), and interestingly, the prevalence and co-infection rate of (up to four species of) Entamoeba was comparably high.
3) Data suggest that IMECs in Western populations are highly reduced compared to rural African populations, but we still need to know more about the relative distribution of for instance fungi and whether these fungi are actually colonising the gut or just carry over from ingested food; right now, it seems as if there might be an inverse relationship between fungal and non-fungal IMECs... something that we can hopefully soon gather sufficient data on for publishing.
4) For those interested in Blastocystis subtype data, including host specificity and geographical distribution, there is a lot to look at in the paper (including supplementary files).

There's a lot more to be said about this paper, but I will sort of leave it here. But please go and read it!


Parfrey, L., Walters, W., Lauber, C., Clemente, J., Berg-Lyons, D., Teiling, C., Kodira, C., Mohiuddin, M., Brunelle, J., Driscoll, M., Fierer, N., Gilbert, J., & Knight, R. (2014). Communities of microbial eukaryotes in the mammalian gut within the context of environmental eukaryotic diversity Frontiers in Microbiology, 5 DOI: 10.3389/fmicb.2014.00298