Showing posts with label Cryptosporidium. Show all posts
Showing posts with label Cryptosporidium. Show all posts

Monday, August 1, 2016

This Month in Blastocystis Research - Interactive Edition

What are your thoughts on Blastocystis carriage and age?

More and more data suggest that the prevalence of Blastocystis carriage increases by age - at least to a certain point.

Some intestinal parasites, such as Cryptosporidium, may not be that uncommon in infants/very young toddlers, while they are much less common in older children and adolescents. Other parasites appear to peak in prevalence around the age of 7, e.g., Dientamoeba fragilis.

Meanwhile, Blastocystis appear to increase in prevalence by age until mature adulthood... why is that? And what does it tell us? Please comment! I'm not having all the answers to these questions myself, and if some interesting suggestions pop up, I'll post them! You only need a Google account to be able to comment. If you don't have one, please send your comment using


For those interested in Blastocystis carriage in association with age, I have listed a couple of relevant recent studies below.


Forsell J, Granlund M, Samuelsson L, Koskiniemi S, Edebro H, & Evengård B (2016). High occurrence of Blastocystis sp. subtypes 1-3 and Giardia intestinalis assemblage B among patients in Zanzibar, Tanzania. Parasites & Vectors, 9 (1) PMID: 27356981  

Poulsen CS, Efunshile AM, Nelson JA, & Stensvold CR (2016). Epidemiological Aspects of Blastocystis Colonization in Children in Ilero, Nigeria. The American Journal of Tropical Medicine and Hygiene, 95 (1), 175-9 PMID: 27139454

Wednesday, December 11, 2013

Molecular Epidemiology: Developing a Language

Initiatives towards standardising diagnostic methods and convening on taxonomy and reference data is extremely important in a world where multiple research teams independently carry out research using molecular markers to identify and differentiate species and genotypes of infectious organisms; such activity is crucial to identify patterns of transmission, differences in virulence, and opportunities for control and intervention. Without such standards, efforts to survey and surveil such organisms would be more or less futile, and so they are the backbone of molecular epidemiology.

Having seen that a variety of morphologically similar but genetically diverse Blastocystis organisms found in humans could in fact colonise a range of different hosts, we realised back in 2006 that all these variants could not all be 'Blastocystis hominis', which was then the species name used for Blastocystis found in humans, and together with colleagues we took to revisiting Blastocystis terminology: We recognised that we did not know enough about host specificity and genetic diversity to be able to come up with relevant species names, and so we invented (or maybe not invented, but at least 'formalised') the subtype system, a sort of a barcode system, where genetically similar (typically 98-100%) organisms are assigned to the same subtype, hence ST1, ST2, ST3, etc., which we today now know so well.

Slapeta now suggests a barcoding system for Cryptosporidium. This single-celled parasite takes a major toll on the health of infants and toddlers in developing countries (in some places surpassed only by norovirus), and may also cause debilitating disease in immunocompromised. The nomenclature for Cryptosporidium is very complicated for those of us who are not experts; for instance, I only recently realised that C. parvum may now only refer to the Mouse I genotype and not the 'common' or 'traditional' C. parvum (which now appears to be C. pestis), which is common in both humans and cattle. However, there is a debate going on as to which taxonomy should be followed, and whether this novel leap in 'Cryptosporidium taxonomy revision' can be endorsed by Slapeta's fellow Crypto experts, remains to be seen. Contentiousness aside, barcoding Cryptosporidium does seem relevant due to the fact that the host specificity of Cryptosporidium is relatively loose; for instance humans and cattle are known to share at least 9 species of Cryptosporidium... 

In his paper, Jan Slapeta lists all the known species of Cryptosporidium (in the 'revised' terminology), and even includes GenBank reference strains for common molecular markers such as actin, HSP70 and COWP1 used for genotyping. Interestingly, he does not include the GP60 marker, a molecular marker for which the terminology is also discordant.

Slapeta moreover includes a file with reference SSU rDNA sequences that enable a standardisation of genetic analyses. This year, we did in fact a similar thing for Blastocystis: Along with our 2013 Protist paper surveying Blastocystis subtypes in animals (including the identification of a couple of new subtypes!), we uploaded a reference alignment consisting of some complete SSU rRNA gene sequences present in GenBank; one or more for each of the now known 17 subtypes; more will be added as more subtypes are discovered. The file can be downloaded when accessing the online version of the paper, and we hope that everyone interested in analysing sequences that represent potentially novel subtypes will use this reference alignment (which has been edited to eliminate regions of ambiguous base alignment); it should be quite helpful. Again, I also bring your attention to the pubmlst Blastocystis database, where fast files obtained by Blastocystis barcoding can be queried in batches for quick analysis of large amounts of sequence data. There's a Youtube video here on Blastocystis barcoding and how to use the pubmlst database.

Consensus on methods, terminology and diagnostic algorithms is essential to developing a common language and understanding of how infectious organisms impact our lives; without it,  confusion wreaks havoc with our efforts.


Alfellani MA, Taner-Mulla D, Jacob AS, Imeede CA, Yoshikawa H, Stensvold CR, & Clark CG (2013). Genetic diversity of Blastocystis in livestock and zoo animals. Protist, 164 (4), 497-509 PMID: 23770574

Kotloff KL, Nataro JP, Blackwelder WC, Nasrin D, Farag TH, Panchalingam S, Wu Y, Sow SO, Sur D, Breiman RF, Faruque AS, Zaidi AK, Saha D, Alonso PL, Tamboura B, Sanogo D, Onwuchekwa U, Manna B, Ramamurthy T, Kanungo S, Ochieng JB, Omore R, Oundo JO, Hossain A, Das SK, Ahmed S, Qureshi S, Quadri F, Adegbola RA, Antonio M, Hossain MJ, Akinsola A, Mandomando I, Nhampossa T, Acácio S, Biswas K, O'Reilly CE, Mintz ED, Berkeley LY, Muhsen K, Sommerfelt H, Robins-Browne RM, & Levine MM (2013). Burden and aetiology of diarrhoeal disease in infants and young children in developing countries (the Global Enteric Multicenter Study, GEMS): a prospective, case-control study. Lancet, 382 (9888), 209-22 PMID: 23680352

Šlapeta J (2013). Cryptosporidiosis and Cryptosporidium species in animals and humans: a thirty colour rainbow? International Journal for Parasitology, 43 (12-13), 957-70 PMID: 23973380  

Stensvold CR, Suresh GK, Tan KS, Thompson RC, Traub RJ, Viscogliosi E, Yoshikawa H, & Clark CG (2007). Terminology for Blastocystis subtypes--a consensus. Trends in Parasitology, 23 (3), 93-6 PMID: 17241816

Striepen B (2013). Parasitic infections: Time to tackle cryptosporidiosis. Nature, 503 (7475), 189-91 PMID: 24236315

Xiao L, Ryan UM, Fayer R, Bowman DD, & Zhang L (2012). Cryptosporidium tyzzeri and Cryptosporidium pestis: which name is valid? Experimental Parasitology, 130 (3), 308-9 PMID: 22230707 

Wednesday, December 4, 2013

This Month in Blastocystis Research (NOV 2013)

Few commercial kits are available for detection of Blastocystis. One of them is the ParaFlorB kit developed by Boulder Diagnostics which uses a monoclonal antibody to detect Blastocystis-specific antigen. We are currently testing this kit in our lab, and I hope to be able to get back with a summary of our experience once the evaluation has finished. Another kit is the EasyScreenTM Enteric Parasite Detection Kit (Genetic Signatures, Sydney, Australia), which was recently evaluated by some of my Australian colleagues (Stark et al., 2013). In this case, Blastocystis has been included in a panel testing for 5 parasitic genera, the other ones being Giardia, Cryptosporidium, Entamoeba, and Dientamoeba, which makes it interesting in a clinical microbiology context, - at least for research purposes.

It can certainly be discussed whether both Dientamoeba and Blastocystis should be part of routine screening for single-celled intestinal parasites. For some years, we have included Dientamoeba in a PCR panel also consisting of Cryptosporidium, Giardia and Entamoeba, but we are about to remove it from this panel. This does not mean that we will not be testing for Dientamoeba; it only means that we will offer testing for Dientamoeba as a separate analysis, in line with our tests for Blastocystis.

According to the study, the kit performs quite well with the only major impediment being the fact that it does not enable differentiation between pathogenic and apathogenic species of Entamoeba; another drawback is the fact that it does not enable detection of the more rare protozoa, such as Cystoisospora and Cyclospora (and I would also mention microsporidia and maybe Balantidium coli). Also, I might be a little worried that the kit will not pick up all species and genotypes of Cryptosporidium, - in fact little was done to challenge the kit in the evaluation. Regarding Cryptosporidium, only C. hominis and C. parvum were tested. In Sweden, at least 10% of all human cryptosporidiosis is due to non-hominis and non-parvum species and genotypes. This is an observation that has led me to revisit our own Cryptosporidium real-time PCR. With help from Welsh and Swedish colleagues I managed to establish quite a broad panel of different Cryptosporidium species and genotypes, and much to my surprise, our 'old' real-time PCR failed to detect the vast majority of these... which means that this Cryptosporidium PCR was far from genus-specific. So, I set out to design a genus-specific PCR which is now being integrated with our Giardia real-time PCR in a duplex assay.
Anyway, similar to Cryptosporidium, many species of Blastocystis - the so-called subtypes - can colonise and infect humans. In the evaluation of the EasyScreen kit, only subtypes 1, 3, and 4 were used to challenge the kit, and so, it is not known whether the kit also detects other subtypes found in humans (ST2, ST5, ST6, ST7, ST8, and ST9).

For those interested in these diagnostic multiplex systems, please also visit a previous blog post.

Anastasios Tsaousis and his Canadian group in Halifax had a paper out just now in Eukaryotic Cell expanding their work on the evolution of the cytosolic iron/sulfur cluster assembly machinery in Blastocystis spp. and other microbial eukaryotes. This type of work is crucial for obtaining a deeper understanding of the metabolism of Blastocystis and to understand how it has evolved and how it potentially differs from other eukaryotes.  Apparently, Iron-sulfur cluster-containing proteins and their biosynthetic machinery in single-celled parasites are remarkably different from those in their mammalian hosts and they therefore represent a potentially relevant target for the development of novel chemotherapeutic and prophylactic agents against parasite infections. For those interested in iron-sulfur clusters in protists in general, a review was published in Advances in Parasitology some weeks ago (please see cited literature).

There is paper out on fasciolosis and co-infections, including Blastocystis, and in that paper it appears that nitazoxanide may be able to eradicate Blastocystis. However, only three persons were treated, and I'm not sure that the diagnostic tests used would have picked up light infections of Blastocystis.

Speaking of treatment: Another paper has appeared from the highly productive team in Sydney, - this time on treatment failure in patients with chronic Blastocystis infection and first-authored by Ms Tamalee Roberts, whom I was so fortunate to spend some time with during the recent congress in Copenhagen. The paper is a little difficult to follow, particularly since nothing is mentioned in the Materials and Methods section on the choice of treatment and treatment strategies in general, but then again, the paper is based on a string of individual (groups of) cases with different kinds of treatment approaches and various backgrounds. I really like the fact that the authors are looking at multiple cases and also that have included a few patients receiving the Triple Therapy (nitazoxanide, furazolidone, secnidazole), which appears to have no major clinical efficacy. The paper also confirms the uselessness of metronidazole when it comes to eradicating Blastocystis. What I could have wished for is that the authors had been able to pursue the microbiological effect of treatment in each of the cases; only in some cases do we get to know about clearance/persistence of Blastocystis. Also, here at the SSI we sometimes wonder, whether persistence of symptoms after treatment may in some cases reflect adverse effects of the treatment (including perturbation of intestinal flora), in which case even randomised controlled treatment (RCT) studies are difficult to design and interpret, unless very clear case definitions and inclusion criteria are available. Hence, for RCT studies I think it is pertinent only to include patients with very similar symptoms (and possibly microbiomes!); given the prevalence of Blastocystis, this shouldn't be too difficult.

Regarding my most recent blog post, I have noticed that it caused quite a stir! I did anticipate some kerfuffle though. But fact is that we have gradually been able to collect so much data from different, independent studies, and the trend appears clear. We now need to investigate what this means, and whether this is something that can be exploited.

There will be no DEC 2013 version of 'This Month in Blastocystis Research' - instead I plan on doing a 'Blastocystis Highlights in 2013' post in line with last year's. Suggestions for significant papers/contributions are welcome!

Cited literature:

Ali V, & Nozaki T (2013). Iron-sulphur clusters, their biosynthesis, and biological functions in protozoan parasites. Advances in Parasitology, 83, 1-92 PMID: 23876871

Roberts T, Ellis J, Harkness J, Marriott D, & Stark D (2013). Treatment failure in patients with chronic Blastocystis infection. Journal of Medical Microbiology PMID: 24243286

Stark D, Roberts T, Ellis JT, Marriott D, & Harkness J (2013). Evaluation of the EasyScreen™ Enteric Parasite Detection Kit for the detection of Blastocystis spp., Cryptosporidium spp., Dientamoeba fragilis, Entamoeba complex, and Giardia intestinalis from clinical stool samples. Diagnostic Microbiology and Infectious Disease PMID: 24286625

Tsaousis AD, Gentekaki E, Eme L, Gaston D, & Roger AJ (2013). Evolution of the Cytosolic Iron/Sulfur cluster Assembly machinery in Blastocystis sp. and other microbial eukaryotes. Eukaryotic Cell PMID: 24243793
Zumaquero-Ríos JL, Sarracent-Pérez J, Rojas-García R, Rojas-Rivero L, Martínez-Tovilla Y, Valero MA, & Mas-Coma S (2013). Fascioliasis and intestinal parasitoses affecting schoolchildren in atlixco, puebla state, Mexico: epidemiology and treatment with nitazoxanide. PLoS Neglected Tropical Diseases, 7 (11) PMID: 24278492