Blastocystis is a genus of single-celled protozoan parasites belonging to a group of organisms

known as the Stramenopiles (also called Heterokonts) that includes algae, diatoms, and water
molds. Blastocystis comprises several species, living in the gastrointestinal tracts of species
as diverse as humans, farm animals, birds, rodents, reptiles, amphibians, fish, and
cockroaches.[1] The species residing in humans is known as Blastocystis hominis and is the
focus of this article.

Blastocystis hominis has a widespread geographic distribution and is found at a rate of 5-10%
in most developed countries, and a rate of up to 50% in less developed areas.[2] High rates of
infection are also found in individuals in developed countries who work with animals.[3]
Although it's role in human disease is frequently referred to as controversial, a systematic
survey of research studies conducted by 11 infectious disease specialists from 9 countries
found that over 95% of papers published in the last ten years identified it as causing illness in
immunocompetent individuals. The paper attributed confusion over pathogenicity to the
existence of asymptomatic carriers, a phenomenon the study noted is common to all
gastrointestinal protozoa.[2]

Classification
The appropriate classification of Blastocystis has only recently been resolved. The original
description of Blastocystis was as a yeast due to its yeast-like glistening appearance in fresh
wet mounts and the absence of pseudopodia and locomotion.[4] This was then contradicted by
Zierdt, who reclassified it under subphylum Sporozoa, based on some distinctive protistan
features of the Blastocystis cell, such as the presence of nuclei, smooth and rough
endoplasmic reticulum, Golgi complex, and mitochondrion-like organelles. Its sensitivity to
antiprotozoal drugs and its inability to grow on fungal media further indicated that it was a
protozoan.

However, major revisions were made to its classification. An analysis of gene sequences was
performed in 1996, which placed it into the group Stramenopiles.[5][6] Other Stramenopiles
include brown algae, mildew, diatoms, the organism that caused the Irish potato famine, and
the organism responsible for Sudden oak death disease. However, the position of Blastocystis
within the stramenopiles remains enigmatic.[7]

Morphology
Blastocystis has various morphological forms.

Four commonly described forms are the vacuolar (otherwise known as central body),
granular, amoeboid, and cyst forms. The appearance of the organism is largely dependent
upon environmental conditions as it is extremely sensitive to oxygen. Whether all of these
forms exist in the host intestine is unclear.

Vacuolar form

The vacuolar form is the typical cell form of Blastocystis seen in culture and is often used for
the identification of the organism. These vacuolar forms vary greatly in size, with diameters
ranging between 2 µm and 200 µm. The vacuolar form is otherwise known as central body
form because it has a large central vacuole surrounded by a thin band of peripheral cytoplasm
which contains other organelles. Flocculent material has been described as being scattered
unevenly throughout the vacuole. The function of the vacuole is still unclear, however, it has
been suggested that, like for many eukaryotic cells, it is for storage purposes. Other functions,
such as cell division during reproduction and the deposition of apoptotic bodies, have been
proposed, although more tests need to be done to validate these roles.

Four common forms of Blastocystis hominis. Clockwise from top left: vacuolar, granular,
amoeboid, and cyst forms.
Granular form

The granular form is somewhat morphologically similar to the vacuolar forms except that
distinct granules are observed in the central vacuole and / or cytoplasm. Within the central
vacuole, these granules appear in different forms too. Three types were suggested – metabolic,
lipid, and reproductive granules. Metabolic granules play a role in chemical processes that are
necessary for the maintenance of life in the organism. It was also put forward that
reproductive granules were involved in the development of progeny cells. These hypotheses
were made based on microscopy alone, which may be deemed misleading, hence more need
to be done before making a definite conclusion. It has also been suggested that the granules
may be an indication that the cell is dying.

Amoeboid form

The other form that exists is the amoeboid form. The amoeboid form of Blastocystis is non-
motile and strongly adhesive. A research study has reported that amoeboid forms are
produced only in cultures taken from symptomatic individuals, with asymptomatic individuals
producing exclusively vacuolar forms. The study suggested this method could be used for
diagnosing symptomatic infection. Additionally, it suggested the symptoms could be due to
the accumulation of the strongly adhesive amoeboid forms on the host's intestinal wall. A
detailed ultra-structural study of amoeboid forms was published in 2007.[16]

Cyst form

The Blastocystis cyst form is a more recent discovery and has helped in the advancement of
understanding the way the infection is transmitted. As compared to the other forms, it is
generally smaller in size and has a thick multilayered cyst wall. It lacks a central vacuole and
few nuclei, multiple vacuoles and food storage deposits were observed. The cyst form is the
most resistant form of this parasite and is able to survive in harsh conditions because of its
thick multilayered cyst wall. Experiments have been carried out to show its ability to
withstand acidic gastric juices. Besides, the cysts did not lyse when placed in distilled water
and could survive well at room temperature for up to 19 days, indicating its strong resistance.
[23][24]
In another experiment, the cyst form was even able to survive in culture medium
containing antiprotozoal drugs. This further supports the idea that the cyst form is the most
resistant of the four forms.

[edit] Life cycle

The supposed life cycle begins with ingestion of the cyst form. After ingestion, the cyst
develops into other forms which may in turn re-develop into cyst forms. Through human
feces, the cyst forms enter the external environment and are transmitted to humans and other
animals via the fecal-oral route, repeating the entire cycle.

Life cycle of Blastocystis proposed by Tan [25]

Obtaining and culturing Blastocystis

The ATCC maintains a collection of Blastocystis isolates. Some records show whether the
isolates were obtained from symptomatic or asymptomatic carriers. As yet, no publication has
identified the subtypes of most of the ATCC isolates, which are mostly axenic. Researchers
have reported that patients with Irritable bowel syndrome may provide a reliable source for
xenic Blastocystis isolates. Some researchers have reported being able to culture Blastocystis
from 46% of IBS patients.[26] Researchers have described different culture mechanisms for
growing Blastocystis. Colony growth on solid medium colonies on solid culture medium using
a synthetic medium with added supplements have both been described.[27][28] However, most
cultivation is performed in liquid media of various type

Mitochondrion-like organelles
The organelles in Blastocystis that resemble mitochondria are an enigma as the organism is a
strict anaerobe. Recent sequence analyses of the organelle genome and over 12,000 expressed
sequence tags (ESTs) has given us many insights into the role these organelles play in the
metabolism of the cell. The genome encodes several subunits of NADH dehydrogenase
(complex I) but lacks all trace of genes for cytochrome and ATPase subunits (Complexes III-
V). ESTs confirm the presence of complexes I and II, and indicate that this partial electron
transport chain may lead to an alternative oxidase. The ESTs also suggest that many other
metabolic pathways characteristic of mitochondria are still present in the Blastocystis
organelles. However, other findings show that the organelle also has characteristics in
common with hydrogenosomes, as a gene encoding [FeFe] hydrogenase is present and the
protein has been localised to the organelles.[29]