Disseminated microsporidiosis: An underdiagnosed and emerging opportunistic disease

@#Disseminated microsporidiosis is a life-threatening disease resulting from the haematogenous spread of microsporidia species. The diagnosis is challenging owing to its subtle nonspecific clinical presentation, which usually reflects the underlying organ involved. Therefore, a high index of suspicion is required for early diagnosis. Besides, tools for confirmatory laboratory diagnosis are limited. Currently, there is no direct diagnostic method that can detect the infection without involving invasive procedures. Clinical confirmation of disseminated microsporidiosis is usually based on light and transmission electron microscopy of infected tissue specimens. These are then followed by species detection using polymerase chain reaction (PCR). Disseminated microsporidiosis shows the potential to be cleared up by albendazole or fumagillin if they are detected and treated early. Based on a series of case reports, this review aims to present a current update on disseminated microsporidiosis with emphasis on the clinical manifestations based on the organ system infected, diagnostic approach and treatment of this devastating condition. Keywords:

Anti-microsporidial effect of thymoquinone on Encephalitozoon intestinalis infection in vitro

To evaluate the anti-microsporidial effects of the active component of Nigella sativa seeds, thymoquinone, against Encephalitozoon intestinalis using an in vitro model. Methods: Anti-microsporidial effect of thymoquinone against Encephalitozoon intestinalis was evaluated by using various concentrations of thymoquinone (0, 1, 5, 10, 15, 20, 30, 35, and 40 uM) and sterile dimethyl sulfoxide. Real time PCR was used to evaluate the inhibitory effects of thymoquinone on the life cycle of Encephalitozoon intestinalis. Results: The cytotoxic effect of thymoquinone on HEK293 cell line was observed with 30, 35, and 40 uM concentrations of thymoquinone after 24, 48, and 72 hours of incubation. It was observed that 10, 15, 20, and 30 uM concentrations of thymoquinone decreased the spore density compared with the control; however, it was significant only at 30 uM. Conclusions: Thymoquinone shows potent anti-microsporidial effects against Encephalitozoon intestinalis in the in vitro model; however, the toxic concentrations of thymoquinone are also toxic to the host cells.

Anti-microsporidial effect of thymoquinone on Encephalitozoon intestinalis infection in vitro

Objective: To evaluate the anti-microsporidial effects of the active component of Nigella sativa seeds, thymoquinone, against Encephalitozoon intestinalis using an in vitro model. Methods: Anti-microsporidial effect of thymoquinone against Encephalitozoon intestinalis was evaluated by using various concentrations of thymoquinone (0, 1, 5, 10, 15, 20, 30, 35, and 40 μM) and sterile dimethyl sulfoxide. Real time PCR was used to evaluate the inhibitory effects of thymoquinone on the life cycle of Encephalitozoon intestinalis. Results: The cytotoxic effect of thymoquinone on HEK293 cell line was observed with 30, 35, and 40 μM concentrations of thymoquinone after 24, 48, and 72 hours of incubation. It was observed that 10, 15, 20, and 30 μM concentrations of thymoquinone decreased the spore density compared with the control; however, it was significant only at 30 μM. Conclusions: Thymoquinone shows potent anti-microsporidial effects against Encephalitozoon intestinalis in the in vitro model;however, the toxic concentrations of thymoquinone are also toxic to the host cells.

Occurrence of zoonotic Enterocytozoon bieneusi in cats in Brazil / Ocorrência de Enterocytozoon bieneusi zoonótico em gatos no Brasil

Abstract Enterocytozoon bieneusi is an opportunistic intestinal pathogen that infects humans and a wide variety of animals worldwide. Our aim in this study was to investigate the occurrence of E. bieneusi in a domestic cat population in Campo Grande, Mato Grosso do Sul, Brazil. Sixty fecal samples from diarrheic cats were subjected to polymerase chain reaction (PCR) and the amplicons were sequenced for identification. E. bieneusi was detected in two samples (3.3%), both identified as genotype D. This genotype has already been reported in animals and humans and is considered a zoonotic genotype. Our findings represent the first report of E. bieneusi in domestic cats in Brazil, reinforcing the importance of identifying this agent as a source of infection in animals and humans.

Resumo Enterocytozoon bieneusi é um patógeno intestinal oportunista que infecta humanos e uma variedade de animais em todo o mundo. O objetivo no presente estudo foi investigar a ocorrência de E. bieneusi em uma população de gatos domésticos em Campo Grande, Mato Grosso do Sul, Brasil. Sessenta amostras fecais de gatos diarréicos foram submetidas a reação em cadeia da polimerase (PCR) e os produtos de amplificação foram sequenciados para identificação molecular. E. bieneusi foi detectado em duas amostras (3,3%), ambos identificados como genótipo D. Esse genótipo tem sido relatado em animais e humanos e é considerado um genótipo zoonótico. Nossos resultados representam a primeira descrição de E. bieneusi em gatos domésticos no Brasil, reforçando a importância desse agente como fonte de infecção para animais e humanos.

Molecular identification of Nosema species in provinces of Fars, Chaharmahal and Bakhtiari and Isfahan (Southwestern Iran)

Objective To molecularly identify Nosema species in provinces of Isfahan, Fars, Chaharmahal and Bakhtiari. Methods One hundred and eighty adult honey bees suspected with nosemosis from provinces of Fars (different counties), Isfahan, and Chaharmahal and Bakhtiari were tested. In order to determine the species of Nosema, previously developed PCR and primers based on 16S rRNA gene were used. PCR products were purified and sent to the Korean company of Macrogen for sequencing. Results Only Nosema ceranae was determined in all samples based on their molecular profile. Sequences of the 16S rRNA gene were sent to GenBank/NCBI (samples accession numbers KP318660–KP318663). Conclusions This species currently exists in European honeybee apiaries of Apis mellifera in the studied provinces.

Microsporidiosis in Iran: A systematic review and meta-analysis

Objective To examine all evidence about Microsporidia infection in vertebrate/invertebrate hosts and Iranian populations distributed in different regions of the country. Methods All published articles up to December 2015, including descriptive and cross-sectional studies related to the prevalence and genotyping of Microsporidia infection in Iran, was considered in this systematic review. The meta-analysis was done using the random-effects model and Stats Direct statistical software. MEGA 5.05 software and maximum likelihood algorithm with Kimura 2-parameter model were used for phylogenetic analysis. Results Of the 1152 investigated studies, 33 eligible studies reported a prevalence of Microsporidia infection in vertebrate and invertebrate hosts. According to this systematic review, the overall prevalence rate of Microsporidia infection in immunocompromised patients in Iran was 8.18%. Furthermore, the overall prevalence rate of Microsporidia infection in immunocompromised patients with chronic diarrhoea, patients with non-diarrhoea, gastroenteritis, and patients with CD4 (<200 cells/μL) was 15.4%, 4.1%, 0.5%, and 12.9% respectively. The highest prevalence rate of human and animal Microsporidia was estimated in Kerman (29%) and Khuzestan (26.5%). The overall prevalence rate of Microsporidia infection in honeybees using the random-effects model was 40%. Furthermore, the highest prevalence rate of nosemosis was described in East Azerbaijan (48.2%). The most Microsporidia isolates from immunocompromised patients and pigeons in Iran belonged to genotypes D (n = 16; 50%) and E (n = 6; 20.6%) of Enterocytozoon bieneusi. Conclusions This study may be the first systematic review and meta-analysis that provides a broad outlook on the prevalence of microsporidiosis in Iran. It is necessary to investigate Microsporidia infection in vertebrate and invertebrate hosts and environmental resources in Iran.

Molecular identification of Nosema species in provinces of Fars, Chaharmahal and Bakhtiari and Isfahan (Southwestern Iran)

Objective: To molecularly identify Nosema species in provinces of Isfahan, Fars, Chaharmahal and Bakhtiari. Methods: One hundred and eighty adult honey bees suspected with nosemosis from provinces of Fars (different counties), Isfahan, and Chaharmahal and Bakhtiari were tested. In order to determine the species of Nosema, previously developed PCR and primers based on 16S rRNA gene were used. PCR products were purified and sent to the Korean company of Macrogen for sequencing. Results: Only Nosema ceranae was determined in all samples based on their molecular profile. Sequences of the 16S rRNA gene were sent to GenBank/NCBI (samples acces-sion numbers KP318660–KP318663). Conclusions: This species currently exists in European honeybee apiaries of Apis mel-lifera in the studied provinces.

Microsporidiosis in Iran: A systematic review and meta-analysis

OBJECTIVE@#To examine all evidence about Microsporidia infection in vertebrate/invertebrate hosts and Iranian populations distributed in different regions of the country.@*METHODS@#All published articles up to December 2015, including descriptive and cross-sectional studies related to the prevalence and genotyping of Microsporidia infection in Iran, was considered in this systematic review. The meta-analysis was done using the random-effects model and Stats Direct statistical software. MEGA 5.05 software and maximum likelihood algorithm with Kimura 2-parameter model were used for phylogenetic analysis.@*RESULTS@#Of the 1152 investigated studies, 33 eligible studies reported a prevalence of Microsporidia infection in vertebrate and invertebrate hosts. According to this systematic review, the overall prevalence rate of Microsporidia infection in immunocompromised patients in Iran was 8.18%. Furthermore, the overall prevalence rate of Microsporidia infection in immunocompromised patients with chronic diarrhoea, patients with non-diarrhoea, gastroenteritis, and patients with CD4 (<200 cells/μL) was 15.4%, 4.1%, 0.5%, and 12.9% respectively. The highest prevalence rate of human and animal Microsporidia was estimated in Kerman (29%) and Khuzestan (26.5%). The overall prevalence rate of Microsporidia infection in honeybees using the random-effects model was 40%. Furthermore, the highest prevalence rate of nosemosis was described in East Azerbaijan (48.2%). The most Microsporidia isolates from immunocompromised patients and pigeons in Iran belonged to genotypes D (n = 16; 50%) and E (n = 6; 20.6%) of Enterocytozoon bieneusi.@*CONCLUSIONS@#This study may be the first systematic review and meta-analysis that provides a broad outlook on the prevalence of microsporidiosis in Iran. It is necessary to investigate Microsporidia infection in vertebrate and invertebrate hosts and environmental resources in Iran.

Identification of opportunistic enteric parasites among immunocompetent patients with diarrhoea from Northern India and genetic characterisation of Cryptosporidium and Microsporidia.

Purpose: Enteric parasitic infestation is a major public health problem in developing countries. Parasites such as Cryptosporidium spp., Cyclospora spp., Cystoisospora spp. and Microsporidia may cause severe diarrhoea among immunocompromised patients. There is scanty data on their frequency among immunocompetent patients. Accordingly, we studied the frequency of enteric opportunistic parasites among immunocompetent patients with diarrhoea from northern India; we also performed genetic characterisation of Cryptosporidia and Microsporidia among them. Patients and Methods: Stool samples from 80 immunocompetent patients with diarrhoea, and 110 healthy controls were examined. Parasites were detected by direct microscopy, modified acid-fast (Kinyoun’s) and modified trichrome stain. Polymerase chain reaction – restriction fragment length polymorphism was used for genetic characterisation of selected species such as Cryptosporidia and Microsporidia. Results: Enteric parasites were detected in 16/80 (20%) patients (mean age 28.8 ± 20 years, 45, 56% males) and in 2/110 (1.8%) healthy controls (P = 0.00007). Parasites detected were Cryptosporidium spp. (8/16, 50.0%), Cystoisospora spp. (4/16, 25%), Microsporidia (1/16, 6.25%), Cyclospora spp. (1/16, 6.25%) and Giardia spp. (1/16, 6.25%). One patient had mixed infection with Cystoisospora spp. and Giardia spp. The species of Cryptosporidia and Microsporidia detected were Cryptosporidium hominis and Enterocytozoon bieneusi, respectively. Parasites were more often detected in younger patients (≤20 years of age) than in older. Most of the parasite infected patients presented with chronic diarrhoea. Conclusion: Opportunistic enteric parasitic infestation was more common among immunocompetent patients with diarrhoea than healthy subjects. Special staining as well as molecular methods are essential for appropriate diagnosis of these parasites.

Intestinal microsporidiosis in renal transplant recipients: Prevalence, predictors of occurrence and genetic characterization.

Purpose: Intestinal microsporidiosis, which occurs in immunocompromised states such as acquired immunodeficiency syndrome, has rarely been studied in patients with renal transplantation (RT) on immunosuppressive therapy. Materials and Methods: Three hundred and twenty‑four consecutive RT recipients on immunosuppressive treatment and 170 healthy subjects were evaluated for intestinal microsporidiosis and other parasites by modified trichrome staining, wet mount using normal saline, iodine and polymerase chain reaction (PCR). Clinical, demographic and laboratory parameters associated with occurrence of intestinal microsporidiosis were studied using univariate and multivariate analysis. The species of microsporidia were studied using PCR‑restriction fragment length polymorphism (RFLP). Patients were treated with albendazole (400 mg twice daily for 2 weeks). Results: Of 324 RT recipients initially screened, 52 were excluded from final analysis due to incomplete data. Patients with RT [n = 272, age 42 ± 12.54 years, 222 (81.6%) male] more often had microsporidiosis than healthy subjects by modified trichrome stain and PCR [n = 170, age 33.8 ± 6.7 years, 123 (72.3%) male] [16/272 (5.8%) vs. 0/170 (0%), P < 0.001]. Patients with intestinal microsporidiosis were younger (33.9 ± 8.3 years vs. 42.3 ± 12.6 years; P = 0.009), had diarrhoea more often (13/16, 81% vs. 123/256, 48%; P = 0.02), which was longer in duration (60, 32.5-105 days vs. 12, 6.2-18 days; P < 0.001) and had associated giardiasis (2/16, 12.5% vs. 2/256, 0.8%; P = 0.018). Younger age, presence of diarrhoea and associated giardiasis were significant on multivariate analysis. Enterocytozoon bieneusi was detected in 15/16 (93%) patients with intestinal microsporidiosis. Conclusion: Intestinal microsporidiosis occurs frequently in patients with RT on immunosuppressive treatment, particularly among younger patients with longer diarrhoea duration and associated giardiasis. E. bieneusi is the major species identified among these patients.

Detection of Encephalitozoon spp. from Human Diarrheal Stool and Farm Soil Samples in Korea

Microsporidia are eukaryotic organisms that cause zoonosis and are major opportunistic pathogens in HIV-positive patients. However, there is increasing evidence that these organisms can also cause gastrointestinal and ocular infections in immunocompetent individuals. In Korea, there have been no reports on human infections with microsporidia to date. In the present study, we used real-time PCR and nucleotide sequencing to detect Encephalitozoon intestinalis infection in seven of 139 human diarrheal stool specimens (5%) and Encephalitozoon hellem in three of 34 farm soil samples (8.8%). Genotype analysis of the E. hellem isolates based on the internal transcribed spacer 1 and polar tube protein genes showed that all isolates were genotype 1B. To our knowledge, this is the first report on human E. intestinalis infection in Korea and the first report revealing farm soil samples as a source of E. hellem infection. Because microsporidia are an important public health issue, further large-scale epidemiological studies are warranted.

Multilocus sequence typing and its application on population genetic struc-ture analysis of parasites / 中国血吸虫病防治杂志

Multilocus sequence typing MLST with high solution sensitivity and specificity is widely used to study the population genetic structure of pathogen by amplification and sequencing of the housekeeping genes. MLST also provides more evidence and plays an important role in parasite research. This paper reviews the principle and method of MLST and its applica-tion on population genetic structure analysis of parasites.

Prevalencia de coccidios y microsporidios intestinales en una comunidad indígena del estado Zulia, Venezuela / Prevalence of Intestinal Coccidia and Microsporidia in an Indigenous Community, State of Zulia, Venezuela

Con la finalidad de determinar la prevalencia de coccidios y microsporidios intestinales, se estudiaron las muestras fecales de individuos de una comunidad indígena de la Sierra de Perijá, Estado Zulia, Venezuela. Para ello, se recolectaron 172 muestras fecales de personas de ambos géneros, con edades comprendidas entre 1 mes y 86 años, a las cuales se les realizaron examen macroscópico y microscópico con solución salina fisiológica, lugol y concentración por formol-éter; del total de muestras solo 145 fueron sometidas a las coloraciones de Kinyoun y Gram-Cromotropo. Dentro de las especies encontradas, se identificó en 3 individuos (2,07%) Cystoisospora belli, Cryptosporidium spp. en 2 (1,38%), Cyclospora cayetanensis en 1 (0,69%) y esporas de Microsporidios en 5 personas (3,45%). La mayoría de los infectados con coccidios y microsporidios intestinales (9/11) presentó asociaciones con otras especies parasitarias, principalmente protozoarios. Un alto porcentaje (54,54%) de los portadores de coccidios y microsporidios, presentó muestras de consistencia diarreica o liquida con moco; tratándose además de personas incluidas en grupos de riesgo (niños y ancianos). Se resalta la necesidad de efectuar el diagnóstico mediante coloraciones especiales para coccidios y microsporidios intestinales, aún en individuos inmunocompetentes y de bajo riesgo, como los indígenas estudiados.

In order to determine the prevalence of intestinal coccidia and microsporidia, fecal samples from individuals of an indigenous community in Perijá, State of Zulia, Venezuela, were studied. One-hundred seventy-two (172) fecal samples from people of both genders between 1 month and 86 years of age were subjected to macroscopic and microscopic examination with physiological saline, Lugol and formalin-ether concentration. Only 145 underwent Kinyoun and gram-chromotrope stains. The following species were identified: Cystoisospora belli in 3 individuals (2.07%); Cryptosporidium spp. in 2 cases (1.38%); Cyclospora cayetanensis in 1 case (0.69%); and microsporidia spores in 5 people (3.45%). Most of those infected with coccidia and intestinal microsporidia (9/11) had associations with other parasitic species, mainly protozoa. A high percentage (54.54%) of the coccidian and microsporidia carriers provided samples of diarrheal and liquid consistency with mucus and were persons included in risk groups (children and elderly). Results emphasize the need for diagnosis by special stains for intestinal coccidia and microsporidia, even in immunocompetent and low-risk individuals, such as the indigenous studied.

Prevalencia de microsporidios intestinales y otros enteroparásitos en pacientes con VIH positivo de Maracaibo, Venezuela / Prevalence of intestinal microsporidia and other intestinal parasites in hiv positive patients from Maracaibo, Venezuela.

Introducción. Desde 1985, los microsporidios se consideran parásitos causantes de infecciones emergentes y oportunistas en individuos inmunocomprometidos en todo el mundo. Objetivo. Detectar la presencia de microsporidios y otros enteroparásitos en pacientes con VIH/sida del Servicio Autónomo Hospital Universitario de Maracaibo (SAHUM), donde no existían estudios previos en este campo. Materiales y métodos. Las muestras fecales se analizaron mediante examen directo, método de concentración con formol-éter, coloración de Kinyoun y coloración Gram-cromotropo rápida. Se realizaron PCR separadas para diferenciar Entamoeba histolytica o Entamoeba dispar , cuando se observó el complejo E. histolytica/dispar al microscopio. Mediante historia clínica se obtuvo información del paciente. Resultados. De los 56 individuos participantes, 38 (67,86 %) presentaron alguna especie parasitaria comensal o patógena en su muestra fecal. Predominaron los individuos portadores de especies parásitas patógenas (26/38). Fueron diagnosticados protozoos como Isospora belli (17,65 %), Blastocystis spp .(17,65 %), Cryptosporidium spp. (7,84 %), complejo Entamoeba histolytica/dispar (5,88 %) , Entamoeba coli (3,92 %) , Giardia lamblia (3,92 %) , Endolimax nana (3,92 %) , Cyclospora cayetanensis (3,92 %) y Chilomastix mesnili (1,96 %). Entre los helmintos, Ascaris lumbricoides, Trichuris trichiura y Strongyloides stercoralis, presentaron un porcentaje de 27,27 % cada uno, e Hymenolepis nana , de 18,18 %. Solo se detectó E. histolytica en uno de los tres casos que presentaron el complejo al examen microscópico. Mediante Gram-cromotropo, 17 muestras evidenciaron esporas del filo Microsporidia, lo que equivale a un 33,33 % de prevalencia. Conclusión. Los microsporidios pueden ocupar el primer lugar de prevalencia en pacientes con VIH positivo, cuando se utilizan técnicas diagnósticas específicas.

Objective: To detect the presence of microsporidia and other enteric parasites in patients with HIVAIDS of the Autonomous Services University Hospital of Maracaibo (SAHUM), where there are no previous studies in this field. Materials and methods: Fecal samples were analyzed by means of direct exam, concetration method with formal-ether, Kinyoun coloration and fast Gram-Chromotrope coloration. Separate PCR were perfomed to differentiate Entamoeba histolytica and Entamoeba dispar , when the E. histolytica/E. dispar complex was observed in the microscope. Information on the patient was obtained trough clinical history. Results: Of 56 individuals that participated, 38 (67.86%) presented some commensal parasite and/ or pathogenic species in their fecal sample. Carriers of pathogenic species were predominat (26/38). Protozoa such as Isospora belli protozoa (17.65%), Blastocystis spp. (17.65%), Cryptosporidium spp. (7.84%), E. histolytica/E. dispar (5.88%), Entamoeba coli (3.92%), Giardia lamblia (3.92%), Endolimax nana (3.92%), Cyclospora cayetanensis (3.92%), and Chilomastix mesnilli (1.96%) were diagnosed. Among the helminths, Ascaris lumbricoides, Trichuris trichiura and Strongyloides stercoralis , had a percentage of 27.27% each, and Hymenolepis nana , 18.18%. Entamoeba histolytica was only detected in one of three cases presenting complex microscopic examination. By Gram-chromotrope, 17 samples showed spores of the Microsporidia phylum, equivalent to 33.33% prevalence. Conclusion: Microsporidia may be first prevalente in HIV positive patients when specific diagnostic techniques are used.

Identificación de Microsporidium spp en pacientes con diagnóstico de leucemia linfoblástica aguda / Identification of Microsporidium spp in patients with acute lymphoblastic leukemia

Introducción. El grupo Microsporidia incluye parásitos intracelulares obligados que afectan varios huéspedes. Los géneros que infectan humanos son Enterocytozoon y Encephalitozoon . Microsporidia es considerado un organismo oportunista en pacientes inmunocomprometidos e inmunocompetentes, que se ha convertido en un importante problema de salud pública. El objetivo del trabajo fue identificar Microsporidium spp en muestras fecales de pacientes con leucemia linfoblástica aguda, en diferente etapa de su tratamiento. Métodos. Se analizaron 77 muestras de niños con diagnóstico de leucemia linfoblástica aguda menores de 12 años, por análisis coproparasitoscópico de Faust. Las muestras se tiñeron con Ziehl Nielsen, tricrómica de Masson y calcoflúor. Finalmente se realizó la amplificación por PCR del gen ribosomal. Resultados. De los pacientes analizados, 16/77 (20.77%) resultaron positivos para Microsporidia, independientemente de que presentaran diarrea. El PCR fue más efectivo para la identificación que el análisis microscópico de las muestras teñidas. Conclusiones. Este trabajo enfatiza la importancia de la microsporidiosis como infección emergente en pacientes con leucemia linfoblástica aguda bajo tratamiento quimioterapéutico, incrementando las complicaciones clínicas adicionales a la leucemia.

Background. The phylum Microsporidia includes obligate intracellular parasites that affect several hosts. The most frequent genera to affect humans are Enterocytozoon and Encephalitozoon. Microsporidium is considered an opportunistic parasite in both immunocompromised and immunocompetent patients and have become an important health problem. The aim of this study was to identify Microsporidium spp in fecal samples of patients with acute lymphoblastic leukemia (ALL), with and without diarrhea, at different treatment stages. Methods. Seventy seven samples from children <12 years old with diagnosis of ALL were analyzed by the Faust coproparasitoscopic method, Ziehl-Nielsen, trichrome and calcofluor staining methods and polymerase chain reaction. Results. Results showed that 16/77 (20.77%) children presented Microsporidium and there was no relationship between microsporidial infection and diarrhea. Polymerase chain reaction was more effective than analysis by light microscopy of staining samples in the identification of the parasite. Conclusions. This work emphasizes the importance of microsporidiosis as an emerging infection in patients with ALL undergoing chemotherapy, increasing the additional clinical complications of leukemia.

Comparison of Calcofl uor White M2R Fluorescence and Modifi ed Gram Chromotrope Kinyoun

Routine diagnosis of intestinal microsporidiosis in clinical diagnostic laboratories relies mostly on detection of microsporidial spores via special staining and microscopic techniques. This paper describes the comparative evaluation of Calcofl uor White M2R method, with modifi ed Gram-chromotrope Kinyoun method as the reference standard. One hundred and six stool samples were examined for the presence of microsporidial spores. Sensitivity, specifi city, positive and negative predictive values of the Calcofl uor White M2R method compared to the reference technique were 95.2%, 4.3%, 78.2% and 20.0%, respectively. The positive predictive value (PPV) was 78.2% and the negative predictive value (NPV) was 20.0%. Despite low specifi city of the CFW method due to its ability to stain chitinous wall of microorganisms, the presence of distinct deep-blue horizontal or equitorial stripes in microsporidial spores in modifi ed Gram-chromotrope Kinyoun would likely reduce the false positive results obtained in the Calcofl uor White M2R. Hence, the simultaneous use of these two methods would give better performance and accuracy for the detection of microsporidial spores in patients with intestinal microsporidiosis.

Microsporidial keratoconjunctivitis in an immunocompetent patient with a past history of laser in situ keratomilieusis surgery.

Ocular infection with microsporidia has been documented in both immunocompetent and immunocompromised patients. Sources and mode of human infection with microsporidia have been difficult to ascertain although exposure to water may be an important risk factor. Of four genera that have been reported in human disease, only the genera Nosema, Encephalitozoon and Septata are documented to cause ocular infection. Here, in our case a healthy 30-year-old man who had undergone bilateral laser in situ keratomilieusis surgery two and half years back presented with a 10-day history of redness and 4-day history of blurring of vision in the right eye. On presentation, his best-corrected visual acuity was 20/20 partial in both eyes. Slit lamp examination revealed multiple pin head shaped infiltrates in the right cornea. Examination of the left eye was unremarkable. Based on microscopic demonstration of numerous microsporidial spores in the corneal scrapings, a diagnosis of microsporidial keratitoconjunctivitis was made. On treatment with oral albendazole, the cornea became clear with complete resolution of symptoms and signs within two weeks.

Characteristics of microsporidial keratoconjunctivitis in an eastern indian cohort: A case series.

Background: Microsporidia are intracellular parasites responsible for human infections. Recently, there has been an increase in the incidence of microsporidial keratoconjunctivitis (MKC) affecting normal individuals worldwide. Aim: To determine the characteristics of MKC in an Indian cohort. Materials and Methods: This is a retrospective, noncomparative, observational case series, involving patients with MKC between June and September 2009. Of the 24 patients identified, microbiological confirmation in direct smear was obtained in 22 cases and selected. Standard microbiological workup was performed in all the cases. We studied the demographics, predisposing conditions, antecedent treatment received before presentation, clinical characteristics, treatment offered, and resolution time with sequel. The management consisted of simple debridement and application of chloramphenicol ointment (1%) two times a day. Results: Mean age of onset was 18.7 years (95% CI, 15.7-21.7; range, 11-36s years). All patients gave history of prior outdoor activity and exposure to rain water/mud. Antecedent treatment comprised of Acyclovir eye ointment (45.4%) and antibiotic eye drop (27.3%) most commonly. Microsporidia were identified in Gram stain (81.8%), 10% potassium hydroxide mount (72.7%), modified Ziehl-Neelsen staining (36.4%), and Giemsa (18.2%). Majority presented as unilateral superficial keratoconjunctivitis with punctate epithelial keratitis. Mean resolution time was 9 days (95%CI, 7.9-10.2). Conclusions: MKC can occur in normal patients with exposure to rain and mud, related to outdoor activity often misdiagnosed as viral ocular infections. Strong clinical suspicion with proper microbiological evaluation helps to diagnose this commonly misdiagnosed condition.

Intestinal microsporidiosis: a hidden risk in rheumatic disease patients undergoing anti-tumor necrosis factor therapy combined with disease-modifying anti-rheumatic drugs?

OBJECTIVE: Immunosuppressed patients are at risk of microsporidiosis, and this parasitosis has an increased rate of dissemination in this population. Our objective was to evaluate the presence of microsporidiosis and other intestinal parasites in rheumatic disease patients undergoing anti-tumor necrosis factor/disease-modifying anti-rheumatic drug treatment. METHODS: Ninety-eight patients (47 with rheumatoid arthritis, 31 with ankylosing spondylitis and 11 with psoriatic arthritis) and 92 healthy control patients were enrolled in the study. Three stool samples and cultures were collected from each subject. RESULTS: The frequency of microsporidia was significantly higher in rheumatic disease patients than in control subjects (36 vs. 4 percent, respectively; p<0.0001), as well as in those with rheumatic diseases (32 vs. 4 percent, respectively; p<0.0001), ankylosing spondylitis (45 vs. 4 percent, respectively; p<0.0001) and psoriatic arthritis (40 vs. 4 percent, respectively; p<0.0001), despite a similar social-economic class distribution in both the patient and control groups (p = 0.1153). Of note, concomitant fecal leukocytes were observed in the majority of the microsporidia-positive patients (79.5 percent). Approximately 80 percent of the patients had gastrointestinal symptoms, such as diarrhea (26 percent), abdominal pain (31 percent) and weight loss (5 percent), although the frequencies of these symptoms were comparable in patients with and without this infection (p>0.05). Rheumatoid arthritis, ankylosing spondylitis and psoriatic arthritis disease activity parameters were comparable in both groups (p>0.05). The duration of anti-tumor necrosis factor/disease-modifying anti-rheumatic drugs and glucocorticoid use were also similar in both groups. CONCLUSION: We have documented that microsporidiosis with intestinal mucosa disruption is frequent in patients undergoing concomitant anti-tumor necrosis factor/disease-modifying anti-rheumatic drug therapy. Impaired host defenses due to the combination of the underlying disease and the immunosuppressive therapy is the most likely explanation for this finding, and this increased susceptibility reinforces the need for the investigation of microsporidia and implementation of treatment strategies in this population.

Parasitosis intestinal en monos capuchinos cariblancos Cebus capucinus (Primates: Cebidae) de un área protegida en la provincia de Limón, noreste de Costa Rica

Intestinal parasites in white-faced capuchin monkeys Cebus capucinus Primates: Cebidae) inhabiting a protected area in the Limón province of Northeastern Costa Rica. Deforestation of tropical forests is threatening monkey biodiversity and their health status, dependent of an ecologically undisturbed area. To asses this relationship, we analyzed parasite occurrence in their intestines. The study was conducted at the Estación Biológica La Suerte (EBLS), Limón, Costa Rica. The group of white-faced capuchin monkeys (Cebus capucinus) was observed between March and December of 2006. A total of 75 feces samples were obtained. Once a sample was collected, the eaten plant type was identified to family and species level, and feces were processed in the laboratory to determine parasite incidence. Results showed that Moraceae was the most represented family in the samples. Among parasites, Strongyloides spp. and Acanthocephala were the most common. Positive prevalence of parasites was found similar and independent of sex and age of capuchin individuals. Microsporids were mainly reported in feces associated with Piperaceae. A low presence of these parasites was found in samples associated with Myrtaceae, with possible anti-parasite active components. The occurrence of parasites was relatively high in EBLS, when compared to other regions in Costa Rica. The higher occurrence of parasites observed in capuchins at EBLS may be due to the fact that this rain forest is surrounded by areas affected by human activities. We suggest the promotion of research in neotropical primates parasitology, for a better comprehension of the parasite-host relationship, and in a long term, being able to understand the ecosystems where they coexist, and consequently, preserve the biodiversity of the whole region. Rev. Biol. Trop. 58 (4): 1335-1346. Epub 2010 December 01.

La deforestación de bosques tropicales está amenazando la biodiversidad de las especies de primates y su estado de salud, dependiente de un lugar ecológicamente equilibrado. Con el fin de evaluar esta relación, se analizó la presencia de parásitos en los intestinos de esta especie. El estudio se realizó en la Estación Biológica La Suerte (EBLS), Limón, Costa Rica. El grupo de monos capuchinos cariblancos (Cebus capucinus) fue observado entre marzo y diciembre de 2006. Se recolectaron 75 muestras de heces a las que se les identificó las plantas ingeridas, y se procesaron en el laboratorio para evaluar la prevalencia de parásitos. Moraceae fue la familia de plantas más reportada. Strongyloides spp. y acantocéfalos fueron los más comunes. La distribución positiva de las mismas fue similar independientemente del sexo y la edad. Microsporidios se reportaron mayoritariamente en heces asociadas con la familia Piperaceae. Fue encontrada una baja existencia de éstos parásitos en muestras asociadas con Myrtaceae en cuyo género Psidium, se han reportado compuestos activos antiparasitarios. La aparición de parásitos fue relativamente mayor en la EBLS, comparado con muestras de otras regiones más secas de Costa Rica. Por lo tanto, esa expresión parasitaria puede deberse al hecho de ser la EBLS una selva lluviosa, además de estar rodeada por zonas con actividades antrópicas. Se sugiere incrementar las investigaciones de parasitología de campo en primates neotropicales con el fin de entender plenamente las relaciones parásitos-hospederos, para en un largo plazo poder comprender los ecosistemas donde conviven, y en consecuencia, preservar la diversidad biológica.