Exacerbated symptoms in Blastocystis sp.-infected patients treated with metronidazole: two case studies.

Blastocystis sp. is a gastrointestinal (GI) protozoan parasite reported to cause non-specific GI symptoms including diarrhea, flatulence, abdominal pain, and nausea. Complete eradication of Blastocystis sp. is rather challenging even with the drug of choice, i.e., metronidazole. Here, we report on two Blastocystis sp.-infected individuals, who presented increased parasite load and exacerbated symptoms upon treatment with the usual recommended dosage and regime of metronidazole. The two studies uniquely demonstrate for the first time a cyst count as high as fivefold more than the original cyst count before treatment and show an exacerbation of GI symptoms despite treatment. The study provides additional support in recognizing metronidazole resistance in Blastocystis sp. and its consequences towards the pathogenicity of the parasite.

PATHOGENICITY OF Blastocystis sp. TO THE GASTROINTESTINAL TRACT OF MICE: RELATIONSHIP BETWEEN INOCULUM SIZE AND PERIOD OF INFECTION / Histopatogenicidade de Blastocystis sp. para o trato gastrointestinal de camundongos: relação com inóculo e tempo de infecção

The pathogenic potential of Blastocystis sp. in experimental models requires further investigation. In this work, the pathogenicity of this parasite in the gastrointestinal tract of male Swiss mice was evaluated according to the inoculum size and period of infection. Animals were infected intragastrically, with 100, 500, 1,000, 5,000 and 10,000 Blastocystis sp. vacuolar forms obtained from a mixture of eight human isolates cultured axenically in Jones' medium. After seven, 14, 21, 28 and 60 days of infection, the animals were sacrificed and fragments of the small intestine (duodenum), large intestine, and cecum were subjected to histopathological analysis. Blastocystis sp. triggered an inflammatory response in the different tissues analyzed, with a predominance of mononuclear cells. The parasite was found in the muscular layer of the cecum, showing its invasive character. Larger inocula triggered inflammatory processes earlier (seven days) than smaller ones (from 21 days). We conclude that, in the proposed model, the pathogenicity of Blastocystis sp. isolates that were studied is related to inoculum size and period of infection.

Pouco é sabido sobre o potencial patogênico de Blastocystis sp. em modelos experimentais. Neste trabalho a patogenicidade desse parasito para o trato gastrointestinal de camundongos Swiss machos foi avaliada de acordo com o inóculo e tempo de infecção. Os animais foram infectados, via intragástrica, com 100, 500, 1.000, 5.000 e 10.000 formas vacuolares de Blastocystis sp. obtidos a partir de uma mistura de oito isolados humanos cultivados axenicamente em meio Jones. Após 7, 14, 21, 28 e 60 dias de infecção os animais foram sacrificados e fragmentos do intestino delgado (duodeno), grosso e ceco foram retirados para análise histopatológica. Blastocystis sp. desencadeou resposta inflamatória nos diferentes tecidos analisados, com predominância de infiltrado mononuclear. No ceco o parasito foi encontrado na túnica muscular mostrando seu caráter invasivo. Inóculos maiores desencadearam processos inflamatórios mais precocemente (7 dias) e inóculos menores mais tardiamente (a partir de 21 dias). Conclui-se que no modelo proposto a patogenicidade dos isolados de Blastocystis sp. estudados tem relação com o inóculo e tempo de infecção.

Stress exacerbates infectivity and pathogenicity of Blastocystis hominis: in vitro and in vivo evidences.

BACKGROUND: Stress alters the oxidant-antioxidant state and immune cell responses which disrupts its function to combat infection. Blastocystis hominis, a common intestinal protozoan has been reported to be opportunistic in immunocompromised patients namely cancer. B. hominis infectivity in other altered immune system conditions especially stress is unknown. We aimed to demonstrate the stress effects towards the susceptibility and pathogenicity of B. hominis infection. METHODS/FINDINGS: Three-week-old Wistar rats were divided into four groups: (a)control; (b)stress-induced; (c)B. hominis infected; (d)stress-induced with B. hominis infection; (n = 20 respectively). Stress was induced for an hour daily (30 days) using a Belly Dancer Shaker. Weight gain was monitored, stool samples were collected for B. hominis screening and blood for the determination of differential count, levels of immunoglobulin, oxidative damage, and peripheral blood mononuclear cell (PBMC) proliferation upon induction with solubilized antigen of B. hominis (Blasto-Ag). Group (b) exhibited the highest level of weight gain. Group (d) had higher levels of parasite cyst count in stools, serum IgE, oxidized protein and lipid compared to the group (c). Levels of monocyte and antioxidant in group (d) were decreased and their PBMCs showed highest inhibition of proliferation level when exposed to Blasto-Ag. Monocyte level in Group (b) showed insignificant difference compared to group (a) but was significantly lower compared to group (c). Antioxidant levels in group (c) were generally lower compared to group (a) and (b). Inhibition level exhibited by Blasto-Ag treated PBMCs of group (c) was higher compared to group (a) and (b). CONCLUSION: The pathogenicity and augmentation of B. hominis infection is enhanced when stress is present. Lifestyles today are becoming increasingly stressed and the present findings suggest that the parasite which has been reported to be one of the most common organisms seen in stool surveys, namely in developing countries, may tend to be more pathogenic in stressful situations.

Clinical significance of Blastocystis hominis: experience from a medical center in northern Taiwan.

BACKGROUND AND PURPOSE: Blastocystis hominis is an intestinal protozoan. The pathogenic role of this organism in human beings is still controversial and has varied among reports from different geographic areas. The purpose of this study was to determine the clinical significance of B. hominis in northern Taiwan. METHODS: A total of 100 patients who had a positive B. hominis stool examination during the period April to December of 2001 were retrospectively identified from Taipei Veterans General Hospital. The demographic and clinical characteristics of these patients were reviewed from the medical records. RESULTS: All of the patients were adults. Fifty nine patients had more than one underlying diseases, including malignancies. Twenty one patients presented with fever and 10 patients had gastrointestinal symptoms, including diarrhea and/or abdominal pain. However, all of the patients had other conditions that might have contributed to the clinical presentation, and they improved without specific treatment for B. hominis. Furthermore, there were no significant differences in clinical symptoms and white blood cell count between patients with malignancy or diabetes mellitus and those without. Six patients had hypereosinophilia that could not be attributed to other causes. Among 34 patients who had a further stool examination within one year, B. hominis was undetectable in 31 patients (91.2%), despite their having no specific antiprotozoal treatment. CONCLUSIONS: The association of clinical symptoms and B. hominis could not be delineated from our study, even in immunocompromised patients. All of the patients improved without receiving any specific therapy. More studies from different areas are needed in order to delineate the clinical significance B. hominis.

Morphology, histochemistry and infectivity of Blastocystis hominis cyst.

Different morphological forms of Blastocystis hominis had been identified in human stool samples. These included both cystic and trophic stages. The latter was induced to encyst by keeping them in potassium dichromate solution for two weeks. Suspected of being the infective stage, cysts were studied in more detail as regards their morphology using both light and electron microscopy. Histochemistry and infectivity studies were also carried out. Light microscopy revealed the cysts to be ovoid or round 5-7 microm with thick cyst wall and a single nucleus. The induced cysts were morphologically similar to the cysts present in the stool. By electron microscope, the cyst wall was evident surrounded by an additional fibrillar coat. The cytoplasm contained one nucleus, many mitochondria, glycogen deposits and a number of variable sized vacuoles. Histochemical studies detected carbohydrates in the cyst wall and fat globules in the cytoplasm. Oral inoculation of albino mice with these cysts led to inflammatory changes in the large and small intestine. The parasite was found at the mucosal epithelium but with no invasion. Different forms of the parasite were detected in the lumen of the intestine.

Blastocystis hominis: parasita intestinal pouco estudado no Brasil / Blastocystis hominis

O objetivo deste artigo é difundir o conhecimento sobre Blastocystis hominis e a doença que produz. O autor apresenta uma revisão começando com os dados históricos e taxonômicos, descrevendo, a seguir, a morfologia, biologia e patogenia do protozoário. Conclui analisando os aspectos clínicos e epidemiológicos, incorporando sua própria experiência no diagnóstico e tratamento desta parasitose intestinal. As considerações realizadas pretendem motivar a realização de pesquisas que permitam determinar as características clínico-epidemiológicas da blastocistose no Brasil

Development of Blastocystis hominis cysts into vacuolar forms in vitro.

The development of cysts of Blastocystis hominis isolated from human feces by the Ficoll-Paque concentration method and cultured in Jones' medium containing 10% horse serum is described. The morphological changes were studied by light and transmission electron microscopy at different intervals for up to 48 h. The cysts developed into a large number of vacuolar forms within 24 h, and binary fission was the only mode of reproduction observed.

Irregular shedding of Blastocystis hominis.

The shedding pattern of the protozoan parasite, Blastocystis hominis, is investigated in man and in experimental animal infections. The shedding pattern of the vacuolar and cystic forms of Blastocystis hominis in infected individuals have been shown in the present study to be irregular. The study shows that there is marked fluctuation in the shedding of the parasite from day to day, varying from as high as 17 to 0 per x40 microscopic field. The cystic stages when estimated in 8 Blastocystis-infected individuals ranged from as high as 7.4x10(5) cysts per gram of stool to 0. The shedding of cystic and vacuolar forms observed over a period of 20 days in experimentally-infected Wistar rats were not only shown to be irregular but the amount varied from host to host. The study has important diagnostic implications in that the stool samples must be collected more than once from patients showing clinical signs and symptoms to eliminate the cause of it to Blastocystis. The study also shows that there are asymptomatic individuals who pass a large amount of cysts as such individuals should be treated to prevent transmission to others.