Saturday, February 23, 2013

Blastocystis aux Enfers

We tremble at the thought of being devoured by a ferocious animal, - of ending our days in a narrow, suffocating slimy tube covered in acidic, nauseating glaze! Remarkably, for some eukaryotic beings, this is the only way forward if they want to carry on with their lives! Intestinal protists such as Blastocystis are in a state of hibernation when outside our bodies and the only thing that may rouse these Sleeping Beauties to action is the passage through low pH enzyme ponds. They thrive, grow and raise their progeny only in the swampy Tartarus of our large intestines; they bequeath to their offspring the affinity for this gloomy, filthy slew; this murky, densely populated, polluted channel, and when the pool of poo becomes all too arid, they know it’s time to buckle up, shut down, and prepare themselves for the great unknown which can potentially mean death to them if eventually they are not lucky enough to be gulped down by another suitable host.

And yet, despite their remarkable modesty and humble requirements these little buggers are being bullied by their inhospitable human hosts; we’d throw anything at them to force them out, organic and inorganic compounds meant to arrest or even kill them. But the whelps of Blastocystis appear extremely resilient, which may hold the key to part of their success; they stay afloat on the Styx of our bowels. In order to eschew Flagyl, perhaps they bribed Phlegyas?

I think it's sometimes useful to put things into a completely different perspective. In any event, from an evolutionary biology standpoint it is highly interesting that a genus which is genetically related to water molds such as those causing potato blight and sudden oak death, has so successfully adapted to a parasitic, anaerobic life style, capable of protractedly colonising a plethora of very diverse host species including members of primates, other mammals, birds, reptiles, amphibians and arthropods and thereby evading innate and adaptive immune defenses from such a diverse range of hosts. One could be inclined to say: Well done! But which is it? Parasitism? Commensalism? Mutalism? Symbiosis? And what will happen to Blastocystis in the future? Will this successful crusader eventually succumb to our avid but maybe imprudent war strategies? And if so, what will happen to us after removing such a common player from our intestinal ecosystems?

Friday, February 22, 2013

Bubbly Blasto!

Yesterday, I was checking up on a fresh Blastocystis culture. I loaded 20 ยตL of the culture "sediment" on to a glass slide, placed the cover slip on top and examined it by light microscopy. While examining the slide, I observed a multitude of dividing cells, indicating vigorous growth and a thriving strain, and once again I was struck by the appearance of dividing Blastocystis. This is basically what they may look like:

Like soap bubbles really, only a lot smaller obviously (mikrons), and somewhat opaque! You'll see them in different sizes and the way they divide looks just like this. Apparently some sort of random budding or multiple fission. You'll see little more than this bubbly structure, which means that there are very few morphological hallmarks to describe. A few nuclei may be discernible along the cytoplasmatic rim, but that's about it when you use light microscopy. Ultrastructural and biochemical analysis is required if you hope to be able to describe some of the processes involved in reproduction.

We often say that Blastocystis organisms representing different subtypes are morphologically indistinguishable; what this actually means is that we do not have the tools to differentiate them morphologically. There may actually be great variation between strains in terms of for instance how they grow in vivo and in vitro and maybe also how they reproduce. Vacuolar forms are the most common form seen in xenic cultures, but other morphotypes are sometimes observed, for instance the granular stage, which, in my experience, is typically seen in cultures that are not “well looked after”, i.e. where medium is not being replaced about twice a week. Dunn and colleagues. (1989) observed that the granular stage could arise from vacuolar stages in cultures where the concentration of horse serum was increased.

I have previously stated that there is no evidence for phagocytosis in Blastocystis. Actually, Dunn et al. (1989) captured what they thought to be bacterial engulfment by ultra-structural analysis, and they also observed bacteria-engulfing pseudopodia in amoeboid stages, in which degraded bacteria were observed. I don't think that I've ever come across this amoeboid stage, but it has been described by quite a few researchers.

Anyway, let's hope for another kind of bubbles this Friday night!

Suggested reading:

Dunn LA, Boreham PF, & Stenzel DJ (1989). Ultrastructural variation of Blastocystis hominis stocks in culture. International Journal for Parasitology, 19 (1), 43-56 PMID: 2707962

Saturday, February 16, 2013

Waiting For The Human Intestinal Eukaryotome

We were lucky enough to have a paper accepted for publication in the ISME Journal (Nature Publishing Group) in which we call for data on the "human intestinal eukaryotome".

In the paper, we start out:

"Recent developments in Next Generation Sequencing (NGS) technologies have allowed culture-independent and deep molecular analysis of the microbial diversity in faecal samples, and have provided new insights into the bacterial composition of the distal gut microbiota. Studies of the microbiome in different patient groups using metagenomics or 16S rRNA gene sequencing are increasing our knowledge of how the microbiota influences health and disease. The majority of recent advances in our understanding of human microbiota structure and dynamic changes in disease were made through phylogenetic interrogation of small subunit (SSU) rRNA (Paliy and Agans 2012). However, until recently such studies have generally failed to include data on common eukaryotic, endobiotic organisms such as single-celled parasites and yeasts ('micro-eukaryotes'). This deficiency may strongly bias the interpretation of results and ignoring an entire kingdom of organisms is a major limitation of human microbiome studies."

Saturday, February 2, 2013

Blog Feedback

I'm very thankful for all the positive feedback I get from readers across the globe, mostly by email. Due to time limits I can only respond to 5-10% of the mail, and I'm sorry for not getting back to the rest of you.

Meanwhile, this blog currently holds more than 60 posts, and you will also find a lot of key words in the right side bar, so take your time and browse a few posts or look up a few relevant key words, -  you might find an answer to one or more  of your questions.

Having said that, I try to read all my email, and I am listening! The feedback and questions that I get are vital for our work and help us identify the avenues that we need to take to unveil the many mysteries of Blastocystis.

And let me just say this for now: A proper microbiological work-up (by state-of-the-art methods, including PCR for intestinal parasites), is something that is offered on a routine basis in only very few laboratories, and also the number of clinically orientated Blastocystis research centres can be counted on one hand, I believe. Subtyping of Blastocystis is currently done mostly in epidemiological surveys (as part of research projects), and I suspect that our lab is one of the very few labs in the world doing subtyping on a routine basis.

Oh, and I've been asked by some readers about how to get blog updates. It's easy: You can follow this blog by email, - just scroll down and find "follow by email" in the right side bar and enter your email address. You can also subscribe to posts via atom (go to the very bottom of the page).

And then here's a little something about stomach acidity and intestinal microbiota from Scientific American, - but make sure to read the comments underneath the post too!