Blastocystis Infection Research Articles

Molecular epidemiology of human Blastocystis in a village in Yunnan province, China

The purpose of this study was to improve our understanding of the molecular epidemiology of human Blastocystis, focusing on 239 randomly selected individuals in a single village in Yunnan province, China. Emphasis was placed on the relative frequency of different Blastocystis subtypes and underlying risk factors. We used a cross-sectional study design, by employing a pre-tested questionnaire to obtain demographic data and behavioural risk factors, and collected faecal samples for culture and subsequent identification of Blastocystis. DNA was extracted from Blastocystis isolates and the subtypes were identified using 7 subtype-specific sequenced-tagged site (STS) primers. Overall, 78 faecal samples were Blastocystis culture-positive (32.6%, 95% confidence interval: 26.7–38.6%). The majority ( n = 73, 93.6%) were single infections with one of the known subtypes, whereas 2 isolates consisted of 2 concurrent subtypes. The remaining 3 isolates could not be identified with the currently known STS primers. Risk factors for a Blastocystis infection were drinking unboiled water, consumption of raw water plants and pig ownership. The consumption of raw water plants was positively associated with subtype 1 infections, and drinking unboiled water with subtype 3 infections. In conclusion, human Blastocystis was common in this village in southwest China, and different subtypes were associated with distinct transmission routes or sources of infection, and hence Blastocystis subtypes might be linked to specific environmental compartments.

Is this recently characterized gastrointestinal pathogen responsible for rising rates of inflammatory bowel disease (IBD) and IBD associated autism in Europe and the United States in the 1990s?

In 2006, a pathogenic variant of the common intestinal organism Blastocystis was discovered in patients who were experiencing chronic gastrointestinal symptoms. Most species of Blastocystis inhabit humans with no symptoms. The discovery of a pathogenic variant of Blastocystis is significant, because Blastocystis is related to Entamoeba, a similar organism with pathogenic variants that kill over 100,000 people each year. Recent research has shown that Blastocystis infections may be undetectable using existing clinical methods. Medical case reports from the Middle East, Europe, and United States suggest that infection with this variant may already be widespread and misdiagnosed as one of several functional disorders. A more virulent or transmissible type of Blastocystis emerged in the Middle East in the 1980's, and was transmitted to Europe and the United States by military and more significantly vacation and business travel. The lack of adequate tests made it impossible to detect the infection. Transmission to the larger population resulted in rising inflammatory bowel disease (IBD) rates in Europe in the 1990's. The relationship between IBD and autism is explored, along with the possibility that the same pathogen causes both conditions. Serological and epidemiological findings are presented supporting the hypothesis. Blastocystis survives sewage treatment, shows low host specificity, and can be spread by many animals. Several communities which have been studied due to high autism rates are located close to rivers which receive large quantities of sewage effluent, such as South Thames (England), Olmsted County (Minnesota, USA) and many communities in Oregon (USA). Scientists from other countries represent the first line of defense against emerging infectious diseases, but their publications on Blastocystis are not well known in the United States and Europe. With the publication of corroborating research by Western scientists in core scientific journals, it is hoped that an appropriate response from the public health system will be forthcoming. Investigation into the existence of infection in the groups mentioned with sensitive and specific tests should be performed. Such tests could include a serum antibody test and a Polymerase Chain Reaction test specific to the pathogenic variant.

Blastocystis: Subtyping isolates using pyrosequencing™ technology

Blastocystis is a prevalent single-celled enteric parasite of unresolved clinical significance. Efforts based on molecular methodologies to establish whether pathogenicity is linked to specific isolates of the genetically diverse genus of Blastocystis have been scarce and so far yielded ambiguous results which can be difficult to interpret. To alleviate some of the problems related to unravelling the molecular epidemiology of Blastocystis infections we developed and evaluated a simple and high-throughput sequence analysis (SQA) pyrosequencing technique based on the detection of genotype-specific nucleotide polymorphisms in the 18S small subunit rRNA gene for a rapid and cost-effective post-PCR screening of Blastocystis genotypes. The method was effectively capable of genotyping 48/48 isolates positive by nested PCR in approximately one hour, and in 94% of the cases the isolate detected by PCR and pyrosequencing was also detected by one of two different PCR assays with subsequent dideoxy sequencing.

Infectivity of Blastocystis isolates from chickens, quails and geese in chickens

The infectivity of six Blastocystis isolates obtained from two domestic chickens, two Japanese quails and two domestic geese, were examined in 1-week-old male chicks. All six isolates were able to infect the chicks via the intracecal inoculation of 1x10(6) cells of cultured organisms. Since the infected chicks discharged many cysts in their feces, the infectivity of the concentrated cysts in chicks was compared among three isolates from different bird species. The CK86-1 and QQ93-3 isolates, which were obtained from a chicken and a quail, respectively, were successfully infected in chicks by orally inoculating with 1x10(2)-1x10(6) cysts. On the other hand, the AC03-1 isolate from a goose required more cysts to infect the chicks, from 1x10(3) cysts to 1x10(6) cysts. In addition, when an uninfected normal chick was housed with five experimentally inoculated chicks with cysts of the QQ93-3 isolate, the normal chick became infected, indicating the fecal-oral transmission of the cyst form among the birds. These results show that the transmission of Blastocystis infection occurs easily between the same or different bird species. Therefore, the proposal of new Blastocystis species on the basis of different avian host species is problematic.

A survey of Blastocystis infection in anuran and urodele amphibians

Blastocystis infection in amphibians was surveyed in three species of anuran and one species of urodele amphibians captured at two distinct locations in Japan. All three species of frogs were highly infected with Blastocystis, while 69 individual urodele newts, Cynopus pyrrhogaster, were negative for infection. Eleven Blastocystis isolates (47.8%) were recovered from 23 Rana nigromaculata leopard frogs. Twenty-three (92%) of 25 Rana catesbeiana bullfrogs and all (100%) of 24 Bufo japonicus japonicus toads were positive for Blastocystis. Two distinct populations of the toad and bullfrog showed a high prevalence (100 or 84.6%) of Blastocystis infection, while in two populations of the leopard frog only one population was positive for Blastocystis (84.6%). Three Blastocystis isolates from different species of the frogs were established. Since none of the three isolates could survive at 37 °C, a temperature tolerance assay was performed to assess the optimal growth temperature and to determine the range of non-lethal temperatures. During the exponential growth phase of 3- or 4-day cultures at 25 °C, three isolates were exposed to 4, 28, 31, or 34 °C for 3 days and then returned to 25 °C to monitor the cell growth. Based on the optimal growth temperatures and different ranges of temperature tolerance among the three new isolates from frogs and two known species, Blastocystis hominis and Blastocystis lapemi, it was established that the three isolates recovered from different species of frogs had different physiological features from B. hominis and B. lapemi.

A survey of Blastocystis infection in anuran and urodele amphibians*1

A survey of Blastocystis infection in anuran and urodele amphibians*1

Blastocystis in animal handlers.

The present study investigated whether people working closely with animals were at higher risk of getting infected with Blastocystis hominis. The prevalence of the parasite was determined in two population groups, i.e., animal handlers and normal healthy individuals who did not work with animals. In all, 105 stool samples were collected from animal handlers from 2 local research institutions, a local zoo, and a local abattoir and 163 stool samples were collected from normal healthy individuals residing in high-rise flats in the city. The in vitro culture method used in the study detected that 41% of 105 animal handlers and 17% of 163 flat-dwellers in the city were positive for Blastocystis. This statistically significant finding (P = 0.0000313) shows that people who work closely with animals do stand at risk of acquiring Blastocystis infection.

A survey of Blastocystis in domestic chickens.

The prevalence and morphology of Blastocystis in fresh faecal material from 227 domestic chickens was investigated. A very high prevalence of infection (approximately 95%) was found in chickens from four of the five commercial farms studied. Extremely high numbers of Blastocystis were found in the majority of samples. Blastocystis cells showed considerable variation in size, ranging from approx. 3 microm to approx. 120 microm in diameter. This size range is more extreme than those previously recognised for the organism from chickens. All chickens from one farm appeared free of Blastocystis infection. Most Blastocystis cells appeared to be the vacuolar form, although the shape of the cells and the appearance of the central vacuole contents varied considerably within and among faecal samples. Nuclei showed "spots" of electron-opaque material, generally arranged as a band within the nuclei. Multiple individual cysts within a single outer fibrillar layer were found in addition to single cysts without an encompassing fibrillar layer.

Severe Blastocystis hominis in an elderly man

We describe a unique case of severe Blastocystis hominis infection in an elderly man with severe dehydration, marked leukocytosis and hypoalbuminaemia after antibiotic treatment for right pneumonia. The patient recovered after treatment with metronidazole. This case presentation demonstrates the ability of B. hominis to induce severe gastrointestinal manifestations and general deterioration, particularly in light of the controversy surrounding its possible potential pathogenicity. We believe, therefore that aggressive treatment with metronidazole should be instituted, following demonstration of the parasite in the stools, if diarrhoea is protracted, since it may well be attributed to Blastocystis infection.

Occurrence of blastocystis sp. in cattle in Aragón, northeastern Spain.

The occurrence of Blastocystis sp. in cattle is reported herein for the first time in Spain. Stool samples from 554 bovines ranging from 3 days old to adults collected from 30 cattle farms in Aragón (northeastern Spain) were concentrated by the formalin-ethyl acetate method and examined light microscopically for the presence of gastrointestinal parasites. Vacuolated cells of Blastocystis sp. was detected in fecal specimens from 10 calves (1.8%) aged from 7 weeks to 6 months. Infected calves were detected at 3 of the 30 farms surveyed (10%). Blastocystis sp. cells were rounded or oval, with a diameter ranging from 6.25 to 15 microns. Blastocystis infection in cattle was never correlated with diarrhea, although infection intensities were generally low.

Use of electron microscopy in examination of faeces and rectal and jejunal biopsy specimens.

The stools and rectal biopsy specimens of 44 patients with AIDS and diarrhoea were examined by culture, light microscopy, and electron microscopy. In 13 patients examination of rectal biopsy material and faecal samples showed no pathogen, but in two of these, microsporidiosis was found by electron microscopical examination of jejunal biopsy specimens. This organism was also identified electron microscopically in one of the further five jejunal biopsy samples taken from patients with a known cause of diarrhoea. Blastocystis hominis infection was identified electron microscopically in six patients, all of whom had cryptosporidiosis additionally seen by light microscopy. Four of these six patients remained well for long periods, with only moderate diarrhoea, and follow up showed no evidence of blastocystis infection. In only four of 11 patients found to have cryptosporidium in their stools at light microscopy were organisms found at electron microscopy. Viral inclusions were only identified at electron microscopy in one of 10 patients with an opportunistic viral infection seen at light microscopy (cytomegalovirus n = 7, herpes simplex virus n = 3). No additional viral pathogens were detected in either stools or rectal biopsy material by electron microscopy. It is concluded that routine electron microscopic examination of stool samples or rectal biopsy material taken from patients with AIDS and diarrhoea is unnecessary and does not increase the yield of potential pathogens compared with standard microbiological techniques and histology.