Low prevalence of Blastocystis sp. in active ulcerative colitis patients.

Ulcerative colitis (UC) is thought to originate from a disbalance in the interplay between the gut microbiota and the innate and adaptive immune system. Apart from the bacterial microbiota, there might be other organisms, such as parasites or viruses, that could play a role in the aetiology of UC. The primary objective of this study was to compare the prevalence of Blastocystis sp. in a cohort of patients with active UC and compare that to the prevalence in healthy controls. We studied patients with active UC confirmed by endoscopy included in a randomised prospective trial on the faecal transplantation for UC. A cohort of healthy subjects who served as donors in randomised trials on faecal transplantation were controls. Healthy subjects did not have gastrointestinal symptoms and were extensively screened for infectious diseases by a screenings questionnaire, extensive serologic assessment for viruses and stool analysis. Potential parasitic infections such as Blastocystis were diagnosed with the triple faeces test (TFT). The prevalence of Blastocystis sp. were compared between groups by Chi-square testing. A total of 168 subjects were included, of whom 45 had active UC [median age 39.0 years, interquartile range (IQR) 32.5-49.0, 49 % male] and 123 were healthy subjects (median age 27 years, IQR 22.0-37.0, 54 % male). Blastocystis sp. was present in the faeces of 40/123 (32.5 %) healthy subjects and 6/45 (13.3 %) UC patients (p = 0.014). Infection with Blastocystis is significantly less frequent in UC patients as compared to healthy controls.

Beyond the association. Toxoplasma gondii in schizophrenia, bipolar disorder, and addiction: systematic review and meta-analysis.

OBJECTIVE: To perform a meta-analysis on studies reporting prevalence of Toxoplasma gondii (T. gondii) infection in any psychiatric disorder compared with healthy controls. Our secondary objective was to analyze factors possibly moderating heterogeneity. METHOD: A systematic search was performed to identify studies into T. gondii infection for all major psychiatric disorders versus healthy controls. Methodological quality, publication bias, and possible moderators were assessed. RESULTS: A total of 2866 citations were retrieved and 50 studies finally included. Significant odds ratios (ORs) with IgG antibodies were found in schizophrenia (OR 1.81, P < 0.00001), bipolar disorder (OR 1.52, P = 0.02), obsessive-compulsive disorder (OR 3.4, P < 0.001), and addiction (OR 1.91, P < 0.00001), but not for major depression (OR 1.21, P = 0.28). Exploration of the association between T. gondii and schizophrenia yielded a significant effect of seropositivity before onset and serointensity, but not IgM antibodies or gender. The amplitude of the OR was influenced by region and general seroprevalence. Moderators together accounted for 56% of the observed variance in study effects. After controlling for publication bias, the adjusted OR (1.43) in schizophrenia remained significant. CONCLUSION: These findings suggest that T. gondii infection is associated with several psychiatric disorders and that in schizophrenia reactivation of latent T. gondii infection may occur.

Eosinophilia à deux: a brain nagging souvenir from the Philippines.

Angiostrongylus cantonensis is the most common cause of eosinophilic meningitis. Although a rare condition among travelers, increased travel and global transportation of food products may result in more cases across non-endemic, developed countries in the future. We here describe two men with headache and painful skin after visiting the Philippines as presenting symptoms. Subsequently, confusion and focal neurologic symptoms developed. Both had increased serum eosinophils; however, CSF eosinophilia was only demonstrated after repeated lumbar puncture. In the CSF of both, Angiostrongylus spp. DNA was detected. Both were treated with albendazole combined with corticosteroids, after which symptoms improved.

Subtype determination of Blastocystis isolates by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS).

The pathogenic role of the enteric parasite Blastocystis remains controversial. Recent studies have suggested that various subtypes (STs) found in human samples could be correlated to the presence or absence and variability of clinical manifestations, and that STs can differ with respect to drug sensitivity. Polymerase chain reaction (PCR) techniques used to determine these STs are expensive and are usually restricted to research laboratory settings. This study evaluates the potential application of the inexpensive matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) technique to discriminate Blastocystis STs. A database of parasitic protein signatures was constructed for five Blastocystis STs, and the reference spectra were challenged with those from 19 axenic cultures of ST1, ST2, ST3, ST4 and ST8 and those from nine xenic liquid cultures of ST3 and ST4. Samples from axenic cultures were prepared using standard formic acid extraction and direct deposition procedures. The reference spectra revealed five distinct spectral profiles, and the database library allowed for discrimination between all of the cultures with reliability indices ranging from 2.038 to greater than 2.8 when an extraction was performed. The direct deposition procedure resulted in greater variability in the discrimination and direct MALDI-TOF MS identification from xenic liquid cultures was effective in 3 out of 9 samples. MALDI-TOF MS proved to be an effective technology for efficiently discriminating Blastocystis STs in axenic cultures.

First case of Echinococcus vogeli infection imported to the Netherlands, January 2013.

In January 2013 in the Netherlands, a man in his 50s from Suriname underwent hemihepatectomy because of a cystic liver mass, assumed to be a cystadenoma. Pathology revealed an echinococcal infection. PCR analysis of cyst material identified Echinococcus vogeli, causing polycystic hydatid disease. This echinococcus species is rarely diagnosed outside South America. The patient received adequate treatment, but this case emphasises the importance of awareness of this infection when treating patients with cystic tumours from endemic areas.

Imported leishmaniasis in the Netherlands from 2005 to 2012: epidemiology, diagnostic techniques and sequence-based species typing from 195 patients.

Leishmaniasis is an imported disease in the Netherlands. We report data for the period between 2005 and 2012, on clinical presentation, country where leishmaniasis was acquired, and causative species, for 195 civilian and military patients who had travelled abroad. Most patients were affected by cutaneous leishmaniasis (CL) (n=185 patients), while visceral leishmaniasis (VL) (n=8 patients) and mucocutaneous leishmaniasis (n=2 patients) were less frequently observed. All VL patients had been infected in Europe. CL was mainly acquired in Afghanistan, Surinam, Morocco and Spain. The majority of CL patients consisted of military personnel (55%, 102/185), 78 of whom had been infected during an outbreak in Afghanistan. Parasitological diagnosis was made by a combination of polymerase chain reaction (PCR), microscopy and culture. Compared to a standard of parasitological proof by any method other than the one under consideration, sensitivities of the individual methods ranged from 73% to 98%. Microscopy was least sensitive, but is fast and cheap. Mini-exon repeat PCR combines high sensitivity and specificity, and allows differentiation between species by sequencing of the PCR product. Eight different species or species complexes were identified, allowing species-specific therapy. Four patients proved infected with Leishmania naiffi, a hitherto rarely described cause of leishmaniasis. In comparison to previous decennia, an increase in cutaneous leishmaniasis was observed in our hospital, both in civilian and military patients who had travelled abroad. This calls for increased awareness among clinicians, availability of diagnostic tests and species-specific treatment guidelines in non-endemic countries.

Chagas disease in European countries: the challenge of a surveillance system.

A study of aggregate data collected from the literature and official sources was undertaken to estimate expected and observed prevalence of Trypanosoma cruzi infection, annual incidence of congenital transmission and rate of underdiagnosis of Chagas disease among Latin American migrants in the nine European countries with the highest prevalence of Chagas disease. Formal and informal data sources were used to estimate the population from endemic countries resident in Europe in 2009, diagnosed cases of Chagas disease and births from mothers originating from endemic countries. By 2009, 4,290 cases had been diagnosed in Europe, compared with an estimated 68,000 to 122,000 expected cases. The expected prevalence was very high in undocumented migrants (on average 45% of total expected cases) while the observed prevalence rate was 1.3 cases per 1,000 resident migrants from endemic countries. An estimated 20 to 183 babies with congenital Chagas disease are born annually in the study countries. The annual incidence rate of congenital transmission per 1,000 pregnancies in women from endemic countries was between none and three cases. The index of under diagnosis of T. cruzi infection was between 94% and 96%. Chagas disease is a public health challenge in the studied European countries. Urgent measures need to be taken to detect new cases of congenital transmission and take care of the existing cases with a focus on migrants without legal residency permit and potential difficulty accessing care.

Miltefosine treatment of Leishmania major infection: an observational study involving Dutch military personnel returning from northern Afghanistan.

In a retrospective, observational study involving 34 patients with Leishmania major infection, 31 of whom had experienced unsuccessful treatment with intralesional antimony (ilSb(v)), miltefosine proved effective. Thirty patients experienced cure after receipt of miltefosine, 3 after receipt of additional ilSb(v), and 1 after 28 daily intravenous injections of antimony. Temporary diminution of ejaculate volume was reported by 21 patients.

[Infestation with the tapeworm Diphyllobothrium latum after eating raw fish]. / Infestatie met de lintworm Diphyllobothrium latum na het eten van rauwe vis.

A 31-year-old man with no relevant medical history encountered a white, ribbon-shaped object, 15 cm long and approximately 1 cm wide, in his faeces. It turned out to be Diphyllobothrium latum, a tapeworm that has fish as the intermediate host. The patient had eaten raw fish and shellfish during a holiday in Brazil 5 months before. He recovered after a single dose of praziquantel.

Diarrhoea in HIV-infected patients: no evidence of cytokine-mediated inflammation in jejunal mucosa.

OBJECTIVE: To determine whether a mucosal cytokine-mediated inflammatory response is involved in cryptosporidial or microsporidial diarrhoea, as well as in diarrhoea of unknown origin in HIV-infected patients. DESIGN: Prospective study. METHODS: Jejunal biopsies were obtained from HIV-infected patients with diarrhoea. Controls were HIV-infected and HIV-seronegative patients without diarrhoea. Two biopsies were homogenized immediately and two other biopsies were first cultured for 20 h. Cytokines [tumour necrosis factor (TNF), interleukin (IL)-1 beta, IL-6, IL-8, IL-10], soluble TNF receptors (sTNFR) p55 and p75, and soluble IL-2 receptor (sIL-2R) were assessed in the homogenates and in the supernatants by sandwich enzyme-linked immunosorbent or enzyme-linked binding assays. The cytokine receptors were also measured in serum. RESULTS: Six HIV-infected patients with cryptosporidiosis, six with microsporidiosis, seven with diarrhoea of unknown origin, seven without diarrhoea, and seven HIV-seronegative patients were eligible. Four patients were excluded because of the presence of other pathogens. No cytokines were detected in immediately homogenized jejunal tissue. Following culture, IL-6 and IL-8 levels were higher in HIV-infected patients with diarrhoea of unknown origin than in HIV-seronegative controls without diarrhoea, although this was not statistically significant. No differences in serum or post-culture supernatant sTNFR p55 and p75 levels existed between the HIV-infected patients with or without diarrhoea. sTNFR, IL-1 beta, IL-10 and the sIL-2R were only detected in low amounts or not at all, and were equally distributed among all patient groups. CONCLUSIONS: This study indicates that mucosal cytokine-mediated inflammatory responses do not play an important role in the pathogenesis of different types of diarrhoea in HIV-infected patients. These results do not support the use of immunomodulatory therapy in these patients.

Improvement of chronic diarrhoea in patients with advanced HIV-1 infection during potent antiretroviral therapy.

BACKGROUND: A substantial number of patients with advanced HIV infection suffer from intractable diarrhoea. The aim of this study was to evaluate whether potent antiretroviral therapy could alleviate such diarrhoea. METHODS: In an open randomized study the effect of the HIV protease inhibitor indinavir in combination with nucleoside analogue reverse transcriptase inhibitors on chronic HIV-related diarrhoea was investigated in 14 late-stage (CD4+ lymphocyte count < or = 50 x 10(6) cells/l) HIV-infected patients. Data concerning stool frequency, stool consistency and antidiarrhoeal drug use were collected in daily diaries over a 24-week period. Endpoints of the study were reduction of stool frequency, improvement of stool consistency, weight gain, and in case of diarrhoea due to Enterocytozoon bieneusi or Cryptosporidium sp. disappearance of these parasites from stool. RESULTS: Thirteen patients started the study drug indinavir. One patient died after 1 week and one patient withdrew prematurely after 18 weeks. Median stool frequency declined from 5.8 daily at baseline to 2.3 daily after 24 weeks (P=0.04). Stool consistency improved considerably over the study period: before treatment 56% of stools were watery and 0% were formed; at week 24 these figures were 0 and 35%, respectively. Body weight increased significantly with a median increment of 6.6 kg at week 24 (P=0.0006). In two out of six patients with microsporidiosis and both patients with cryptosporidiosis, stools were free of parasites at week 24. Five out of six patients who used non-specific antidiarrhoeal medication on a regular basis prior to the study had ceased to do so at the end. CONCLUSION: The use of potent antiretroviral therapy in patients with advanced HIV infection can improve chronic HIV-related diarrhoea and in some cases lead to disappearance of E. bieneusi and Cryptosporidium sp. from the stools.

Pyrimethamine alone as maintenance therapy for central nervous system toxoplasmosis in 38 patients with AIDS.

We retrospectively assessed the efficacy of maintenance therapy with pyrimethamine alone in 38 patients with AIDS and central nervous system (CNS) toxoplasmosis. The diagnosis was based on clinical presentation and compatible CT scan abnormalities with subsequent response to therapy. Survival analysis was performed by the product limit method of Kaplan-Meier. Fourteen patients received maintenance therapy with 25 mg pyrimethamine per day (group 1), and 24 patients were treated with 50 mg per day (group 2). The median survival from initiation of maintenance therapy until death or end of the study for the entire study population was 32 weeks. Median survival in group 1 was 28 weeks, as compared with 36 weeks in group 2 (p = 0.34). Relapses occurred in 12 patients, six in group 1 and six in group 2. There was no significant difference in failure-free survival between the two treatment groups (p = 0.09). One patient in group 1 and two patients in group 2 experienced severe toxicity, requiring discontinuation of therapy. All three patients relapsed and died. Two patients in group 2 who stopped treatment on their own initiative also had relapses. Thus, all five patients who discontinued therapy had relapses. Five of 13 patients in group 1 and two of 20 patients in group 2 relapsed during continuous therapy with pyrimethamine (p = 0.13); these seven patients responded to reintroduction of combination therapy (n = 6) or treatment with 50 mg pyrimethamine per day (n = 1). The results of our retrospective analysis suggest that maintenance therapy with oral pyrimethamine, 50 mg per day, in AIDS patients with CNS toxoplasmosis is effective.

Diagnosis of intestinal and disseminated microsporidial infections in patients with HIV by a new rapid fluorescence technique.

AIMS: To assess the value of a new rapid fluorescence method for the diagnosis of microsporidiosis in HIV seropositive patients. METHODS: Microsporidian spores in stools were demonstrated by using the fluorochrome stain Uvitex 2B. The new technique was evaluated in three groups of HIV seropositive patients with diarrhoea. Group 1: 19 patients with biopsy confirmed E bieneusi infection (186 stool samples); group 2: 143 consecutive patients from whom faeces were submitted for routine investigation of diarrhoea (318 samples); group 3: 16 patients with small intestinal biopsy specimens negative for microsporidia (55 samples). The new method was used to monitor spore shedding during experimental treatment with paromomycin and albendazole in four patients. RESULTS: Brightly fluorescent spores were detected in all stool samples of patients in group 1. In group 2 16 (11%) patients had spores in their stool samples. E bieneusi was found in 11 patients; in the other five another genus of microsporidia, Encephalitozoon, was recognised. Encephalitozoon spores were also found in the urine of three of these patients and in the maxillary sinus aspirate of two of them, suggesting disseminated infection. The results were confirmed by electron microscopic examination. In group 3 negative biopsy specimens were confirmed by negative stool samples in all cases. Treatment with albendazole and paromomycin did not affect the spore shedding in three patients with E bieneusi infection. By contrast, in a patient with Encephalitozoon sp infection albendazole treatment resulted in clinical improvement together with complete cessation of spore excretion in the stool. CONCLUSION: The Uvitex 2B fluorescence method combines speed, sensitivity, and specificity for the diagnosis and treatment evaluation of intestinal and disseminated microsporidiosis.

Dissecting the IgM antibody response during the acute and latent phase of toxoplasmosis.

A major problem in anti-toxoplasma IgM serology is the occurrence of clinically non-relevant (CNR) IgM in sera of latently infected (LI) individuals. The susceptibility for CNR IgM of the Toxo ISAGA IgM, Platelia Toxo IgM and Vidas Toxo IgM assays was determined using sera of LI individuals with an IgM titer in the Abbott IMx Toxo IgM assay. The specificity of CNR-IgM and IgM antibodies in sera of acutely infected (AI) individuals was compared by immunoblotting.Seven/19 samples with CNR IgM antibodies were found positive in the ISAGA IgM, compared with 16/19 and 17/19 with the Vidas and Platelia Toxo IgM assays, respectively. In contrast, immunoblotting allowed a distinction between CNR IgM and AI IgM antibodies of the 30 sera studied.Clearly, IgM assays are susceptible to CNR IgM antibodies. In cases of doubt, immunoblotting is of value as confirmation method. Because CNR IgM antibodies are specific for toxoplasma, it will be difficult to improve IgM ELISAs in such a way that CNR IgM antibodies will not interfere with the analysis.

Serum neopterin concentrations during treatment of leishmaniasis: useful as test of cure?

Neopterin, a product of gamma-interferon-activated macrophages, was measured in sera from 28 patients (12 patients with cutaneous leishmaniasis and 16 patients with visceral leishmaniasis) to determine the utility as a marker of disease activity and therapeutic efficacy. Patients originated from Kenya (n=5) and from the Academic Medical Center, Amsterdam, The Netherlands (n=23). In seven patients follow-up sera after treatment were available. Two patients at the time of diagnosis of visceral leishmaniasis were co-infected with HIV. The 12 patients with cutaneous leishmaniasis had serum neopterin levels below the upper limit of the normal range. All 16 patients with visceral leishmaniasis had elevated levels of serum neopterin before treatment. In six out of seven patients with visceral leishmaniasis followed during treatment neopterin levels decreased to values below the upper limit of the normal range (10 nmol l(-1)). Sequential measurements of serum neopterin levels may be useful for monitoring therapeutic efficacy in patients with visceral leishmaniasis.

High prevalence of Enterocytozoon bieneusi infections among HIV-positive individuals with persistent diarrhoea in Harare, Zimbabwe.

Infection with the microsporidian parasite Enterocytozoon bieneusi may be a major cause of prolonged diarrhoea in individuals also infected with human immunodeficiency virus (HIV). The parasite has been reported from Europe, Australia and the Americas, with a prevalence of 7-29%. Faecal specimens were obtained from 202 adults and 106 children in Harare, Zimbabwe, all of whom were in hospital and had diarrhoea. HIV serology was available for 119 adults: 106 were HIV seropositive. There were clinical grounds for suspecting HIV infection in 23 of the remaining patients. E. bieneusi was identified in specimens from 13/129 patients (10%) for whom HIV infection was indicated by serology and/or clinical signs, 1/60 patients (2%) of uncertain HIV status, and 0/13 seronegative patients. 18/106 children were HIV seropositive and 12 were not; HIV serology was not available for the remainder, but 19 were strongly suspected of being infected with HIV on clinical criteria. E. bieneusi was not detected in samples from any child. As is common in Zimbabwe, the prevalence of other parasites in faecal specimens was low and, amongst patients with proven or suspected HIV infection, E. bieneusi was the most prevalent parasite identified, particularly in patients with diarrhoea of over 4 weeks duration.

Evaluation of the Eiken latex agglutination test for anti-Toxoplasma antibodies and seroprevalence of Toxoplasma infection among factory workers in Addis Ababa, Ethiopia.

Sera from 170 factory workers aged 18-45 years enrolled in a pilot study of human immunodeficiency virus 1 (HIV-1) infection in Addis Ababa, Ethiopia, were screened for anti-Toxoplasma immunoglobulin G antibodies by the Sabin-Feldman test (reference standard) and the Eiken latex agglutination test (under evaluation for use in developing countries). Based on the Sabin-Feldman test, the prevalence of anti-Toxoplasma antibodies was 80.0% (95% confidence interval 73.9-86.1%). The sensitivity and specificity of the Eiken latex agglutination test were 96.3% and 97.1%, respectively, showing its validity for the detection of anti-Toxoplasma antibodies. The prevalence of antibodies did not differ between individuals infected and uninfected with HIV-1 (74.2% versus 83.3%, P > 0.05). However, antibody titres were higher in HIV-infected persons than in those who were uninfected (P < 0.001). Based on these findings, we expect that toxoplasmic encephalitis will be a common opportunistic infection among HIV-infected Ethiopians, and chemoprophylaxis with co-trimoxazole may be beneficial to those with low CD4+ T cell counts. The prognostic significance of high titres of anti-Toxoplasma antibodies remains to be established among Ethiopian HIV-infected individuals.

Electron microscopy as an essential technique for the identification of parasites in aids patients.

We use EM with increasing frequency for the identification of opportunistic parasitic infections in HIV-infected individuals. Apart from Pneumocytis carinii, Toxoplasma, Cryptosporidium, and Leishmania, we studied several aspects of microsporidiosis. Infection with the intestinal microsporidian Enterocytozoon was found in as much as 27% of AIDS patients with chronic diarrhoea without other pathogens. EM diagnosis of microsporidiosis is commonly performed on intestinal biopsies, but we have recently demonstrated spores of microsporidium with a non-invasive technique, viz. in faeces (1). However, EM of biopsy material remains the reference technique to distinguish the various species. Combining faeces and biopsy examination, we identified another group of microsporidians, Encephalitozoon sp., in the small intestine of AIDS patients with chronic diarrhoea (Fig. 1). Encephalitozoon sp. with identical ultrastructure was found in urine and sinus discharge, suggesting dissemination of the infection. In the maxillary sinus of one patient, we demonstrated E. bieneusi, a parasite which had previously been found only in small intestine and bile duct epithelium (2) (Fig. 2). After albendazole treatment, Encephalitozoon sp. disappeared from faeces, urine and nasal discharge. Although ultrastructural damage was noted in the developmental cycle of E. bieneusi in biopsies after treatment with albendazole, spores continued to be present in the faeces. These results demonstrate the great value of EM in the diagnosis of several parasitic diseases, especially microsporidiosis.

[3 emerging protozoal infections in The Netherlands: Cyclospora, Dientamoeba, and Microspora infections]. / Drie opkomende protozoaire infectieziekten in Nederland: Cyclospora-, Dientamoeba- en Microspora-infecties.

Increased international travelling, an increased number of patients with immunosuppression caused by HIV infection, and renewed interest in known but little studied microorganisms, resulted in a more frequent finding of certain medically important parasites. Three emerging protozoal infection, Cyclospora cayetanensis, Dientamoeba fragilis and Microspora (Enterocytozoon bieneusi and Encephalitizoon) are causative agents of diarrhoea. Encephalitozoon infections are also associated with hepatitis, hepatoconjunctivitis and nephritis. C. cayetanensis infection was diagnosed in 28 patients in the years 1992-1995 in the Academic Medical Centre in Amsterdam, half of these patients returning from a visit to Indonesia. D. fragilis has a prevalence in the Netherlands of 8% among patients with diarrhoea lasting longer than one week referred for parasitological investigation. The prevalence of E. bieneusi in HIV positive patients with diarrhoea was 8-10% in the Academic Medical Centre in the last five years. Infection with Encephalitozoon appears to be endemic in the Netherlands.