In vitro Effect of Silver Nanoparticles on Blastocystis hominis.

INTRODUCTION: This study aims to assess the efficacy of silver nanoparticles (Ag Nps) alone and combined with metronidazole (Ag Nps + MTZ) as potential alternative therapeutic agents for Blastocystis hominis. METHODS: The parasites were challenged with Ag Nps, Ag Nps + MTZ and MTZ. To assess the efficacy of drugs, counting of viable parasites was done after 1, 2, and 3 hours of adding the drugs. RESULTS: Blastocystis hominis count was reduced by 20.72%, 28.23%, and 18.92% after one hour of adding Ag Nps, Ag Nps + MTZ, and MTZ, respectively. Cysts count was further reduced by 51.49%, 61.61%, and 40.78% after 2 hours and by 71.69%, 79.67%, and 62.65% after 3 hours of adding the drugs in the same order, respectively. CONCLUSION: There was a statistically significant difference (P<0.05) in the in vitro growth inhibition of the parasite over the different time intervals when using the tested drugs against the control drug.

Low efficacy of metronidazole in the eradication of Blastocystis hominis in symptomatic patients: Case series and systematic literature review. / Escasa eficacia de metronidazol en la erradicación de Blastocystis hominis en pacientes sintomáticos: serie de casos y revisión sistemática de la literatura.

INTRODUCTION: Blastocystis hominis (B. hominis) is a protozoan commonly found in the gastrointestinal tract. There are doubts about its clinical significance. Metronidazole (MTZ) is the recommended first-line treatment. MATERIALS AND METHODS: A retrospective review was carried out between 2011 and 2012. A total of 151 samples were randomly selected from 383 samples positive for B. hominis. Inclusion criteria were: suggestive symptoms, treatment indication and microbiological follow-up. A systematic review was performed of all studies that evaluated the effect of MTZ on B. hominis infection. RESULTS: Forty-six patients met the inclusion criteria (64% women; age, 44.2±2 years). MTZ was used in 39 patients, 31 of whom obtained a clinical response (79.5%) but only 15 a microbiological response (48.4%). No dose-effect relationship was observed. Twenty patients with no initial microbiological response received a second round of treatment (MTZ, cotrimoxazole, paramomycin, others), with a microbiological response in 70%. Overall, B. hominis was cured in 72% (95% CI: 57%-83%). Of 54 treatments associated with a clinical response, a microbiological response occurred in 31 (57%), while in the remaining 12 with no clinical response, microbiological cure was observed in only 2 (17%) (P=.022). The eradication rate in the systematic review varied between 0% and 100%. CONCLUSIONS: There seems to be a relationship between the clinical and microbiological response to B. hominis treatment. The microbiological response to MTZ treatment is insufficient in our geographical setting. The systematic review shows that the response to MTZ is very variable.

[Frequency and in vitro susceptibility antiparasitic of Blastocystis hominis from patients admitted to the Hospital Regional Lambayeque, Peru]. / Frecuencia y susceptibilidad antiparasitaria in vitro de Blastocystis hominis en pacientes admitidos en el HospitalRegional Lambayeque, Perú.

OBJECTIVE: To describe the frequency and antiparasitic in vitro susceptibility of Blastocystis hominis in patients admitted to theHospital Regional Lambayeque, Peru. MATERIAL AND METHODS: A cross-sectional study was conducted from January to August 2015 at 313 patients of all ages. B. hominis detection was performed on serial fecal samples by direct microscopic examination and microculture in modified Locke solution. The in vitro susceptibility testing against the drug metronidazole, nitazoxanide, trimethoprim-sulfamethoxazole and erythromycin was performed in 24 strains of B. hominis, which grew up (microculture method) in 10 double concentrations of each antimicrobial (from 256 ug/ml to 0.5 ug/mL) plus a control. RESULTS: 46.3% (145/313) of the sample had B. hominis, also the age between 12 to 17 years and 60 years was associated with higher frequency of parasites (OR: 2.93 and 2.62). The minimum inhibitory concentration (MIC) 90 of metronidazole and nitazoxanide was 3.19 ug/mL and 11.19 ug/ml, respectively, whereas the MIC 90 of trimethoprim-sulfamethoxazole and erythromycin were above 256 ug/mL. CONCLUSIONS: B. hominis occurs in high frequency in patients admitted to the Hospital Regional in Lambayeque, proving to be an important problem of public health in the region. Also B. hominis isolated from these patients were shown to be susceptible in vitro to low concentrations of metronidazole and nitazoxanide so they could be chosen for treatment of this parasite.

IN VIVO AND IN VITRO EFFICACY OF NIGELLA SATIVA AQUEOUS EXTRACT ON BLASTOCYSTIS HOMINIS.

Metronidazole (MTZ) was the most widely accepted treatment for Blastocystis hominis (B. hominis) with high treatment failure rate, resistance and potential mutagenic and carcinogenic effects so there is urgent need to find out new, effective and safe treatment against B. hominis. The present research aimed to evaluate the therapeutic effect of the aqueous extract of Nigella sativa (NS) at different doses on B. hominis in vitro and in vivo in comparison to MTZ as a control drug. Isolates of B. hominis were obtained from patients complaining of diarrhea and abdominal pain. Isolates were cultured in egg diphasic medium (LE) for in vitro study and to adjust proper inoculating dose for in vivo study. The aqueous extract of NS at concentrations of 100 & 500 µg/ml showed a potent lethal effect on B. hominis isolates in vitro. Caecal tissue of experimentally infected and treated mice with two different doses of NS (250 & 500 mg/kg/d) were examined histopathologically and compared with that of mice infected and treated by two doses of MTZ (62 & 125 mg/kg/d) as control drug and Infected untreated mice as negative control group. Histopathological examination of infected untreated group showed all pathological degrees in the caecal tissue while infected treated one showed remission of pathological changes especially with higher dose (500 mg/kg). Present study proved that N. sativa had inhibitory effect on B. hominis in vitro and prevented cytopathic effect in infected mice inoculated orally with B. hominis.

In vitro effect of some Egyptian herbal extracts against Blastocystis hominis.

Blastocystis hominis is an enteric parasite that inhabits the gastrointestinal tract of humans and many animals. This emerging parasite has a worldwide distribution. It is often identified as the most common eukaryotic organism reported in human fecal samples that showed a dramatic increase in recent years. Metronidazole is the main therapy for blastocystosis. However, frequent reports of treatment failure suggesting isolates resistance to metronidazole. This study determined the growth pattern and in vitro susceptibility of B. hominis to nitazoxanide (NTZ), garlic, ginger, onion and turmeric. Fecal samples positive for Blastocystis were collected from patients with irritable bowel syndrome (IBS), and processed for culture. Cultured samples were subjected to examination by light microscopy. Herbs' extracts was freshly prepared. Drug susceptibility assays was done using 0.1 mg/ml of NTZ, garlic, ginger, onion and turmeric. Effects assessed on parasite culture after 24 hr. and 48 hr. Cultured fecal samples of B. hominis have identified several forms of the organism; vacuolar, granular, amoeboid and cyst forms within 24 hr. Nitazoxanide treatment significantly (P < 0.001) lowered the parasite number after 48 hr. (mean, 337.5 ± 17.67) /ml. The reduction rate after 48 hr. compared to PBS was 93.33%. Ginger treatment significantly (P < 0.002) lowered the number of the parasite after 48 hr. (mean, 335 ± 7.07)/ml. Moreover, garlic treatment also significantly (P < 0.002) lowered the number of the parasite after 48 hr. (mean, 382.5 ± 10.60)/ml. The reduction rates after 48 hr. in these treated samples compared to PBS were 92.98% and 92.44% respectively. However, onion, and turmeric treatments insignificantly lowered the number of the parasite after 48 hr. (P < 0.15 & < 0.22 respectively).

Protozoa as a cause of recurrent abdominal pain in children.

OBJECTIVES: The aim of this study was to investigate whether protozoa can be identified as a cause of recurrent abdominal pain (RAP), and whether protozoan infections can be recognized by a specific clinical presentation. METHODS: For 2 years, all patients (ages 4-16 years) fulfilling the Apley criteria of RAP referred to secondary care were prospectively evaluated for protozoa (Giardia lamblia, Dientamoeba fragilis, Blastocystis hominis) and treated if positive. Re-examination followed at least 10 days after treatment. Disappearance of pain with eradication and a pain-free follow-up of at least 6 months were considered to be indicative of a causal relation with RAP. The predictive value of the characteristics of the pain for protozoan infections was calculated. RESULTS: Of 220 included patients (92 boys, mean age 8.8 years), 215 brought a stool sample; 73 (34%) carried parasites, 10 of whom had 2 parasites, 2 had 3 parasites. Sixty-five patients were treated. Twenty-five (11%) were pain-free after eradication (21 had D fragilis, 8 B hominis, 4 G lamblia), of whom 11 had another infection (2) or constipation (9) as second diagnosis for the pain. Five had recurrence of infection with D fragilis and were again pain-free with eradication. Patients with protozoa as cause of their pain did not show differences with respect to their presentation when compared with patients with an asymptomatic infection and patients without protozoa. CONCLUSIONS: Protozoa were found as the cause of pain in 6% to 11% of children with RAP. These patients did not show a characteristic presentation when compared with patients with other causes of abdominal pain.

No advantage for antibiotic treatment over placebo in Blastocystis hominis-positive children with recurrent abdominal pain.

OBJECTIVE: The aim of the study was to investigate whether recurrent abdominal pain (RAP) in Blastocystis hominis-positive children can be treated successfully with trimethoprim-sulfamethoxazole (TMP/SMX). METHODS: From October 2004 to December 2008, all of the patients referred to the Division of Gastroenterology and Nutrition of the University Children's Hospital Zurich because of RAP and detection of B hominis in stool samples as the only pathological finding after a standard workup were offered to participate in the study. Patients were randomly assigned into 2 groups. TMP/SMX or placebo was given for 7 days in a double-blind, placebo-controlled manner. Pain index (PI) was measured with a visual analogue scale. Two weeks after completion of treatment, 3 stool samples were collected and patients were followed clinically. If B hominis was still present, metronidazole was given for 7 days. RESULTS: Forty patients were included; 37 finished the study (TMP/SMX n = 20, placebo n = 17). Mean PI declined from 7.1 to 3.6 for all of the patients, with a decrease from 6.9 to 4.1 in the TMP/SMX and 7.4 to 3.0 in the placebo group, irrespective of detection of B hominis after treatment. There was no statistically significant difference in PI reduction between the 2 groups. Metronidazole treatment led to a further PI decline from 3.7 to 1.9. Eradication rates were 35% (TMP/SMX) and 44% (metronidazole), compared with spontaneous clearance of 29% in the placebo group. CONCLUSIONS: There is no advantage for TPM/SMX over placebo in the treatment of RAP in B hominis-positive children.

In vitro sensitivity of Blastocystis hominis to garlic, ginger, white cumin, and black pepper used in diet.

To determine the growth pattern and in vitro susceptibility of Blastocystis hominis to metronidazole (MTZ), garlic, ginger, white cumin, and black pepper. Stool specimens were collected from 16 irritable bowel syndrome (IBS) and 10 controls between July-November 2010. Stool microscopy and culture for B. hominis was performed. Drug susceptibility assays was done using 0.01 and 0.1 mg/ml of MTZ, garlic, ginger, white cumin, and black pepper. Effect was assessed on B. hominis culture after 48 h. Stool DNA was extracted using stool DNA extraction kit (Qiagen) and polymerase chain reaction (PCR) done using subtype-specific sequence-tagged-site primers. B. hominis genotype 3 and coinfection of 1 and 3 tended to grow well in culture compared to isolated type 1 infection. Exposed to MTZ at a concentration of 0.01 mg/ml, 38% (6/16) B. hominis from IBS did not grow in culture compared to 100% (10/10) of B. hominis from control (p = 0.001). When they were exposed to MTZ at 0.1 mg/ml, 56% (9/16) B. hominis from IBS did not grow in cultures compared to 100% (10/10) from control (p = 0.01). Forty-four percent (7/16) B. hominis from IBS did not grow in culture compared to 100% (10/10) B. hominis from control when exposed to garlic at a concentration of 0.01 mg/ml (p = 0.003) and following exposure to garlic at 0.1 mg/ml, 38% (6/16) B. hominis from IBS did not grow in cultures compared to 100% (10/10) from control (p = 0.001). B. hominis isolates from IBS had a cell count of 6,625 at a MTZ concentration of 0.01 mg/ml that reduced to 1,250 as MTZ concentration was increased to 0.1 mg/ml (p = 0.08). B. hominis from IBS with a mean cell count of 3 × 10(5) at baseline decreased to 1 × 10(4) when exposed to garlic at 0.01 mg/ml (p < 0.001) and to 1 × 10(3) (p < 0.001) when garlic was 0.1 mg/ml. B. hominis from IBS cell count decreased to 1 × 10(5) when exposed to white cumin at 0.01 mg/ml (p = 0.01) and to 1 × 10(5) (p < 0.001) when white cumin was 0.1 mg/ml. Exposed to black pepper at 0.1 mg/ml, cell count of B. hominis from IBS decreased to 1 × 10(5) (p = 0.01). B. hominis from IBS decreased to 1.3 × 10(5) exposed to ginger at 0.01 mg/ml (p = 0.001). B. hominis isolates were mostly genotypes 3, type 1 and 3 coinfection, and non-typeable B. hominis isolates. B. hominis isolates from IBS mostly genotype 1 demonstrated an increased sensitivity to garlic at 0.01 mg/ml with a B. hominis cell count of 3,714 compared to 6,142 when exposed to 0.01 mg/ml of MTZ. However, this sensitivity did not increase as garlic concentration was increased to 0.1 mg/ml, for B. hominis cell count was 6,000 compared to 1,428 as MTZ was increased to 0.1 mg/ml.

Clinical efficacy of Saccharomyces boulardii or metronidazole in symptomatic children with Blastocystis hominis infection.

Although many Blastocystis infections remain asymptomatic, recent data suggest it also causes frequent symptoms. Therapy should be limited to patients with persistent symptoms and a complete workup for alternative etiologies. The goal of this study was to compare the natural evolution (no treatment) to the efficacy of Saccharomyces boulardii (S. boulardii) or metronidazole for the duration of diarrhea and the duration of colonization in children with gastrointestinal symptoms and positive stool examination for Blastocystis hominis. This randomized single-blinded clinical trial included children presenting with gastrointestinal symptoms (abdominal pain, diarrhea, nausea-vomiting, flatulence) more than 2 weeks and confirmed B. hominis by stool examination (B. hominis cysts in the stool with microscopic examination of the fresh stool). The primary end points were clinical evaluation and result of microscopic stool examination at day 15. Secondary end points were the same end points at day 30. Randomization was performed by alternating inclusion: group A, S. boulardii (250 mg twice a day, Reflor®) during 10 days; group B, metronidazole (30 mg/kg twice daily) for 10 days; group C, no treatment. At day 15 and 30 after inclusion, the patients were re-evaluated, and stool samples were examined microscopically. On day 15, children that were still symptomatic and/or were still B. hominis-infected in group C were treated with metronidazole for 10 days. There was no statistically significant difference between the three study groups for age, gender, and the presence of diarrhea and abdominal pain. On day 15, clinical cure was observed in 77.7% in group A (n, 18); in 66.6% in group B (n, 15); and 40% in group C (n:15) (p < 0.031, between groups A and C). Disappearance of the cysts from the stools on day 15 was 80% in group B, 72.2% in group A, and 26.6% in group C (p = 0.011, between group B and group C; p = 0.013, between group A and group C). At the end of the first month after inclusion, clinical cure rate was 94.4% in group A and 73.3% in group B (p = 0.11). Parasitological cure rate for B. hominis was very comparable between both groups (94.4% vs. 93.3%, p = 0.43). Metronidazole or S. boulardii has potential beneficial effects in B. hominis infection (symptoms, presence of parasites). These findings challenge the actual guidelines.

[The effect of trimethoprim-sulfamethoxazole in Blastocystis hominis infection]. / Blastocystis hominis infeksiyonunda trimethoprim-sülfametoksazolün etkisi.

The aim of this study was to investigate clinical findings and the effects of trimethoprim-sulfamethoxazole (TMP-SMX) in cases of blastocystosis. A total of 37 cases whose stool specimens were sent to the parasitology laboratory from the outpatient clinics of our hospital for various reasons were included in the study. Only five or more Blastocystis hominis were found during examination with direct wet mount using the 40x objective. The stool specimens were tested for other agents (Salmonella spp., Shigella spp., Escherichia coli H157:07, rotavirus) and cases with one of these were excluded from the study. The cases with blastocystosis were given TMP-SMX for 7 days. After the treatment, the cases were questioned as to symptoms once again, the stool specimen examinations were repeated with the same methods, and the results were evaluated. In 34 (91.89%) out of the 37 cases where B. hominis was found, various clinical symptoms such as stomach ache, flatulence, diarrhea, itching and fever were observed singly and/or together. After the treatment it was found that 36 (97.3%) out of 37 cases improved. This study supports the premise that TMP-SXT is effective in the treatment of B. hominis.

Evaluation of the nitric oxide activity against Blastocystis hominis in vitro and in vivo.

The effect of exogenous nitric oxide (NO) on growth, viability and ultra-structural of B. hominis was assessed in vitro by sodium nitrite (NaNO2) in 0.6 mM, 0.8 mM & 1 mM concentrations. The viability of B. hominis was identified using neutral red stain. The role of NO as an endogenous oxidant was assessed by identifying its level in cecum tissue, ileum tissue, blood and stool elutes of mice infected with B. hominis symptomatic human isolates using reactive nitrogen assay compared to control. In vitro study revealed that NaNO2 inhibited the growth and decreased viability of B. hominis with minimal lethal concentra-tion dose 1 mM on the 4th day while, minimal effects were detected with 0.6 and 0.8 mM. Transmission electron microscopy study proved that apoptotic-like features were observed in growing axenic culture of B. hominis upon exposure to NaNO2. These changes were not only found on the vacuolar (central body) form but also they were detected on granular, multi-vacuolar and cyst forms. In vivo study proved that high levels of NO were found in infected mice compared to low changes in control group. The high levels were in cecum tissue particularly. The mean levels of NO among infected mice were 211.8 +/- 20.7 microM in cecum, 90.4 +/- 11.6 microM in ileum, 60.1 +/- 4.7 microM in blood and 63.6 +/- 7.3 microM in stool elutes while, the mean levels of NO in control mice were 70.2 +/- 3.1 in cecum, 67.8 +/- 4.7 microM in ileum, 30.9 +/- 4.2 microM in blood and 28.1 +/- 2.9 microM in stool elutes. The differences were statistically highly significant. NO-donor drugs proved useful in treatment and increase the host resistance to B. hominis.

Effect of exogenous adminstration of antioxidants on Blastocystis hominis in vivo.

The effect of exogenous administration of antioxidant (Anttox) on the course of B. hominis in experimentally infected mice was studied. B. hominis isolates were obtained from 10 gastrointestinal symptomatic adult patients. Three groups of 30 infected mice (3/isolate) were used. GI was untreated infected, GII was treated by antox for 4 weeks after infection diagnosis (treatment strategy), and GIII antox treated by for 4 weeks before infection (prophylactic strategy). Mild pathological changes were detected on 13.4%, 19.9% & 86.8% of mice in Gs I, II & III, respectively. Moderate pathological changes were found in 29.9%, 26.6% & 6.6% of mice in Gs I, II & III, respectively. While, the majority of severe pathological changes were in Gs I & II (56.7% & 53.5%) as compared to GIII (6.6%). Meanwhile, 86.8% of mice in GIII had B. hominis forms > 10/high power field compared to 3.3% in Gs I & II, respectively. Although 19.8% of mice in GII were positive for B. hominis by direct smear, no growth resulted in vitro and all the forms were non-viable by using neutral red stain. All the differences were statistically significant. So, antioxidant exacerbated B. hominis intensity but it decreased the pathological changes.

Pathogenicity of Blastocystis hominis, a clinical reevaluation.

Blastocystis (B.) hominis was considered to be a member of normal intestinal flora in the past, but in recent years it has been accepted as a very controversial pathogenic protozoan. In this study, 52 individuals whose stool examination revealed B. hominis were evaluated for clinical symptoms. Metronidazole was administered for 2 weeks to the patients infected with B. hominis. After 2 weeks of treatment they were called for a follow-up stool examination. No other bacteriological and parasitological agents were found during stool examination of these patients. The frequency rate of intestinal symptoms was 88.4% in the B. hominis cases. Abdominal pain was the most frequent symptom (76.9%). Diarrhea and distention followed at a rate of 50.0% and 32.6%. Intestinal symptoms may be seen frequently together with the presence of B. hominis and this protozoan may be regarded as an intestinal pathogen, especially when other agents are eliminated.

The use of the antifungal agent miconazole as an inhibitor of Blastocystis hominis growth in Entamoeba histolytica/E. dispar cultures.

In regions with high prevalence, Blastocystis hominis is frequently found in association with Entamoeba histolytica/E. dispar in xenic cultures. Its exacerbated growth is often superimposed on the growth of amebas, thus impeding the continuation of the amebas in the culture, within a few generations. The present study reports on the excellent efficacy (100%) of the antifungal agent miconazole in eliminating B. hominis from cultures of E. histolytica/E. dispar, thereby maintaining the integrity of the trophozoites of the amebas. Nystatin presented low efficacy (33.3%).

The use of the antifungal agent miconazole as an inhibitor of Blastocystis hominis growth in Entamoeba histolytica/E. dispar cultures

In regions with high prevalence, Blastocystis hominis is frequently found in association with Entamoeba histolytica/E. dispar in xenic cultures. Its exacerbated growth is often superimposed on the growth of amebas, thus impeding the continuation of the amebas in the culture, within a few generations. The present study reports on the excellent efficacy (100 percent) of the antifungal agent miconazole in eliminating B. hominis from cultures of E. histolytica/E. dispar, thereby maintaining the integrity of the trophozoites of the amebas. Nystatin presented low efficacy (33.3 percent).

Em regiões de alta prevalência, Blastocystis hominis é freqüentemente encontrado em associação com Entamoeba histolytica/E. dispar em cultivos xênicos. Seu crescimento exacerbado se sobrepõe muitas vezes ao das amebas, impedindo a manutenção destas em cultura, dentro de poucas gerações. O presente estudo relata a excelente eficácia (100 por cento) do antifúngico miconazol na eliminação de B. hominis dos cultivos de E. histolytica/E. dispar, mantendo-se a integridade dos trofozoítos das amebas. A nistatina apresentou eficácia baixa (33,3 por cento).

Evaluation of the in vitro effect of Nigella sativa aqueous extract on Blastocystis hominis isolates.

The effect of Nigella sativa aqueous extract was evaluated against the in vitro growth of 2 different isolates of the intestinal protozoan parasite Blastocystis hominis. Different concentrations (10, 100, 500 microg/ml) of Nigella aqueous extract and metronidazole, an active standard drug for B. hominis, were incubated with B. hominis isolates in culture media at 37 degrees C. Their possible effect on B. hominnis living cell count (LCC) was assessed on Day 1, 3 & 6. The aqueous extract of N. sativa at concentrations of 100 and 500 microg/ml showed a potent lethal effect on both B. homninis isolates, but with different extent. There is no significant difference between the inhibitory effect of N. sativa and metronidazole on the LCC on the 6th day. On assessment of living cell rate (LCR) which calculate percentage rate of living cell, N. sativa at 500 microg/ml concentration has a significant inhibitory effect on both isolates. So, it is considered as the most active concentration of Nigella aqueous extract. These results prove that N. sativa aqueous extract could be useful in the treatment of B. hominis.

Protease activity of Blastocystis hominis.

Parasite-derived proteases are important for the parasite life cycle and the pathogenesis of the disease they produce. Proteases of intestinal protozoan parasite Blastocystis hominis were studied for the first time with azocasein assays and gelatin SDS-PAGE analysis. Parasitic lysates were found to have high protease activity and nine protease bands of low (20-33 kDa) and high (44-75 kDa) molecular weights were reported. Proteases were found to be pH-dependent and highest proteolytic activity was observed at neutral pH. Inhibition studies showed that B. hominis isolate B, like many other protozoan parasites, contains mainly cysteine proteases.

Blastocystis hominis and the evaluation of efficacy of metronidazole and trimethoprim/sulfamethoxazole.

Blastocystis hominis is commonly found in the intestinal tract of humans. Although the pathogenicity of this unicellular parasite is controversial, anti-protozoan agents are usually administered to infected individuals. At present, the first choice of chemotherapeutic agent is Metronidazole as described in the literature. In this study, we evaluated the effects of metronidazole and Trimethoprim/Sulfamethoxazole (TMP/SMX) on persons infected with B.hominis. A total of 104 subjects infected with B. hominis were admitted to the laboratory from 2002 to 2003. All individuals were non-immunocompromised and subjects were monitored for 1 year after treatment. All stool samples were microscopically examined after staining with iodine and by culturing in an egg slant medium. Of the 104 infected individuals (52+/-16 years of age, M:F=60:44) with B. hominis infection, 28 were discharging large numbers of parasites before treatment. Of 28 severely infected individuals, 12 were treated with metronidazole/250-750 mg at a regimen of 3 x/day/10 days and 4 of the 12 were eradicated. Nine individuals were treated with TMP/SMX/1 tab at a regimen of 3 x/day/10 days and 2 of the 9 were eradicated. For severe B. hominis infections, it appears that metronidazole and TMP/SMX are effective in some individuals, but not all.

Caspase-3-like protease influences but is not essential for DNA fragmentation in Blastocystis undergoing apoptosis.

Blastocystis hominis undergo apoptosis after treatment with a cytotoxic monoclonal antibody (MAb), 1D5, by mechanisms that are not fully understood, although our previous study demonstrated that caspase-3-like protease activity is involved. To elucidate the mechanism of MAb 1D5-induced apoptosis, we inhibited Blastocystis caspase-3-like protease to investigate if there would be a concomitant decrease in in situ DNA fragmentation. However, MAb 1D5-induced apoptosis, evidenced by DNA fragmentation, was not completely blocked by pretreating with specific caspase-3 inhibitor, Ac-DEVD-CHO, indicating that caspase-independent apoptotic pathways might also be involved. Our results also revealed that the treatment with MAb 1D5 resulted in the loss of mitochondrial membrane potential (deltapsim), independent of Ac-DEVD-CHO pretreatment. In conclusion, this study demonstrates that MAb 1D5-induced apoptosis in B. hominis is not wholly dependent on caspase-3-like protease activity and is associated with mitochondrial dysregulation. This is the first report showing evidence for complex apoptotic pathways in a unicellular parasite.

In vitro susceptibility of Blastocystis hominis isolated from patients with irritable bowel syndrome.

This study aims to determine the growth pattern and in vitro susceptibility of clinical isolates of Blastocystis hominis to different concentrations of metronidazole, furazolidone and ciprofloxacin. Stool specimens from 25 consecutive patients with irritable bowel syndrome presenting to the gastroenterology department of Aga Khan University Hospital between January and May 2003 are examined by microscopy and cultured for B. hominis. Drug susceptibility assays are performed for metronidazole, furazolidone, and ciprofloxacin using final concentrations of 0.01 mg/mL and 0.1 mg/mL. The effect of the drugs is assessed after B. hominis culture for 48 h. With furazolidone and metronidazole, 68% (17/25) and 60% (15/25) of B. hominis isolates, respectively, failed to grow at drug concentrations of both 0.01 mg/mL and 0.1 mg/mL. However, ciprofloxacin failed to suppress growth completely at both concentrations. B. hominis resistance to furazolidone, metronidazole and ciprofloxacin at 0.01 mg/mL was 32% (8/25), 40% (10/25) and 100% (25/25), respectively. B. hominis isolates varied in their degree of susceptibility to the three drugs studied, being greater with furazolidone than with metronidazole, and complete resistance with ciprofloxacin.