Microscopio and PCR-based detection of microsporidia spores in human stool samples

Abstract Microsporidia are obligate intracellular fungi with a remarkable ability to infect a wide range of invertebrate and vertebrate hosts. Namely, Enterocytozoon bieneusi is the most frequently microsporidia reported worldwide, and mainly associated with chronic diarrea and wasting syndrome in AIDS patients. Microscopy and PCR-based detection techniques are effective for diagnosis and identification of species and genotypes; however, these methods should be standardized in each laboratory. In this study, we performed microscopy and nested PCR techniques with PCR product sequencing to detect E. bieneusi in human stool samples. These techniques, if applied together, might prove useful for diagnosis and future epidemiological studies of intestinal microsporidiosis in Argentina.

Resumen Los microsporidios son hongos intracelulares obligados con una notable capacidad para infectar una amplia gama de hospedadores invertebrados y vertebrados. Enterocytozoon bieneusi es el microsporidio más frecuentemente reportado en todo el mundo, principalmente tricrómicaasociado con diarrea crónica y síndrome debilitante en pacientes con sida. Las técnicas dedetección basadas en microscopía y PCR son útiles para el diagnóstico y la identificación deespecies y genotipos, pero estos métodos deben estar estandarizados en cada laboratorio.En este estudio evaluamos técnicas de microscopía y PCR anidada, con secuenciación de losproductos, para detectar E. bieneusi en muestras de heces humanas. Estas técnicas, usadas con-juntamente, podrían ser útiles para su aplicación en el diagnóstico de microsporidiosis intestinaly para realizar estudios epidemiológicos de esta afección en Argentina.

Prevalence of Enteric Protozoan Oocysts with Special Reference to Sarcocystis cruzi among Fecal Samples of Diarrheic Immunodeficient Patients in Iran

The genus Sarcocystis is not usually considered as an important enteric pathogen in immune compromised patients. It might be expected that species for which humans are the final host (Sarcocystis hominis and Sarcocystis suihominis as well as possibly others) would be encountered increasingly often in immunodeficient persons. This study aimed to address how to detect and differentiate Sarcocystis oocysts and/or sporocysts from enteric protozoans in the diarrheal samples of immunodeficient patients in Shiraz, Iran. Diarrheal samples of 741 immunodeficient patients with recurrent persistent or chronic diarrhea were examined by microscopy and molecular biological analysis. Oocysts-positive samples were 68 Cryptosporidium spp., 9 Cystoisospora belli (syn. Isospora belli), 2 Cyclospora cayetanensis, and 15 microsporidia (Enterocytozoon bieneusi). Sarcocystis-like sporocysts found from a woman were identified as Sarcocystis cruzi through 18S rDNA amplification and phylogenetic analysis. To the best of our knowledge, this is the first report of S. cruzi from a human.

Description of microsporidia in simulids: molecular and morphological characterization of microsporidia in the larvae of Simulium pertinax Kollar (Diptera: Simuliidae)

Introduction Microsporidia constitute the most common black fly pathogens, although the species' diversity, seasonal occurrence and transmission mechanisms remain poorly understood. Infections by this agent are often chronic and non-lethal, but they can cause reduced fecundity and decreased longevity. The objective of this study was to identify microsporidia infecting Simulium (Chirostilbia) pertinax (Kollar, 1832) larvae from Caraguatatuba, State of São Paulo, Brazil, by molecular and morphological characterization. Methods Larvae were collected at a single point in a stream in a rural area of the city and were kept under artificial aeration until analysis. Polydispyrenia spp. infection was characterized by the presence of at least 32 mononuclear spores measuring 6.9 ± 1.0 × 5.0 ± 0.7µm in persistent sporophorous vesicles. Similarly, Amblyospora spp. were characterized by the presence of eight uninucleate spores measuring 4.5 × 3.5µm in sporophorous vesicles. Results The molecular analysis confirmed the presence of microsporidian DNA in the 8 samples (prevalence of 0.51%). Six samples (Brazilian larvae) were related to Polydispyrenia simulii and Caudospora palustris reference sequences but in separate clusters. One sample was clustered with Amblyospora spp. Edhazardia aedis was the positive control taxon. Conclusions Samples identified as Polydispyrenia spp. and Amblyospora spp. were grouped with P. simulii and Amblyospora spp., respectively, corroborating previous results. However, the 16S gene tree showed a considerable distance between the black fly-infecting Amblyospora spp. and the mosquito-infecting spp. This distance suggests that these two groups are not congeneric. Additional genomic region evaluation is necessary to obtain a coherent phylogeny for this group. .

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.

Microsporidial spores can cross the intact descemet membrane in deep stromal infection

We report a rare case of a deep stromal keratitis with a chronic indolent course, diagnosed as microsporidial keratitis from corneal scrapings. The patient's condition worsened despite medical therapy and penetrating keratoplasty was performed. The histopathology of the corneal tissue revealed multiple microsporidial spores in the posterior stroma and the endothelial exudates, whereas there was no clinical or histopathological breach in Descemet's membrane. This is the second report in the literature to report that micropsoridial spores can cross the intact Descemet's membrane

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.

Multiplex PCR Detection of Waterborne Intestinal Protozoa; Microsporidia, Cyclospora, and Cryptosporidium

Recently, emerging waterborne protozoa, such as microsporidia, Cyclospora, and Cryptosporidium, have become a challenge to human health worldwide. Rapid, simple, and economical detection methods for these major waterborne protozoa in environmental and clinical samples are necessary to control infection and improve public health. In the present study, we developed a multiplex PCR test that is able to detect all these 3 major waterborne protozoa at the same time. Detection limits of the multiplex PCR method ranged from 101 to 102 oocysts or spores. The primers for microsporidia or Cryptosporidium used in this study can detect both Enterocytozoon bieneusi and Encephalitozoon intestinalis, or both Cryptosporidium hominis and Cryptosporidium parvum, respectively. Restriction enzyme digestion of PCR products with BsaBI or BsiEI makes it possible to distinguish the 2 species of microsporidia or Cryptosporidium, respectively. This simple, rapid, and cost-effective multiplex PCR method will be useful for detecting outbreaks or sporadic cases of waterborne protozoa infections.

Ocorrência de microsporídios em pequenos mamíferos silvestres no Estado de São Paulo / Occurrence of Microsporidia in small wildlife mammals in the state of São Paulo, Brazil

This study describes infection of microsporidia in the natural fauna of small wildlife animals in an area of deforestation for a water reservoir construction in the state of São Paulo, Brazil. It was focused on marsupials, small rodents, frogs, primates, bats, and others mammals taken from this area to access whether they may represent environmental sources of these zoonoses. From all captured animals, microsporidia spores were observed in the feces of bats, small rodents, and marsupials. This study emphasises the importance of wild animals, particularly small mammals as potential sources of microsporidia to human and animal populations from deforested areas.

Ocorrência de Giardia, Cryptosporidium e microsporídios em animais silvestres em área de desmatamento no Estado de São Paulo, Brasil / Occurrence of Giardia, Cryptosporidium and microsporidia in wild animals from a deforestation area in the state of São Paulo, Brazil

A ocorrência de Giardia, Cryptosporidium e microsporídios foi investigada por meio da análise de 98 amostras fecais de animais silvestres capturados em uma área de desmatamento para a construção das barragens de Paraitinga e Biritiba, localizadas nos Municípios de Mogi das Cruzes, Salesópolis e Biritiba-Mirim, no Estado de São Paulo. As amostras foram obtidas de 46 roedores, 21 marsupiais, 16 sapos, nove morcegos, três primatas e três lagartos. As técnicas de centrífugo-flutuação com sulfato de zinco, de Kinyoun e a coloração de Gram-Chromotrope foram utilizadas, respectivamente, para a pesquisa de Giardia, de Cryptosporidium e de microsporídios. O total de animais parasitados por um dos protozoários investigados foi de 17,35 por cento (17/98). Cistos de Giardia foram encontrados em amostras fecais de dois pequenos roedores da espécie Coendou villosus (ouriço-cacheiro). Os três animais positivos para Cryptosporidium foram roedores das espécies Akodon montensis, Thaptomys nigrita (ambos conhecidos como ratos do mato) e Sciurus aestuans (serelepe ou caxinguelê). Esporos de microsporídios foram encontrados nas fezes de 12 animais, sendo seis roedores das espécies Oligoryzomys sp.(um), Akodon montensis (três) e Coendou villosus (dois), três marsupiais pertencentes às espécies Didelphis aurita (dois) e Marmosops incanus (um) e três morcegos da espécie Diphylla ecaudata. Este é o primeiro relato de microsporidiose em animais silvestres no Brasil. A presente investigação enfatiza a importância de animais silvestres, particularmente pequenos mamíferos, como potenciais fontes de infecção desses protozoários para outras populações animais, incluindo o homem, em áreas de desmatamento.

The occurrence of Giardia, Cryptosporidium and microsporidia was investigated in 98 faecal specimens from wildlife animals, captured in an area of deforestation for the construction of two water reservoirs (Paraitinga and Biritiba), located in the municipalities of Mogi das Cruzes, Salesópolis and Biritiba-Mirim, in the state of São Paulo (Brazil). Samples were obtained from 46 rodents, 21 marsupials, 16 frogs, 9 bats, 3 tamarins and 3 lizards. For the detection of Giardia, Cryptosporidium and microsporidia it was used, respectively, the floatation technique with lead sulphate, the Kinyoun method and the Gram-Chromotrope staining. The total number of parasitized animals by one of these protozoans was 17.35 percent (17/98). Cysts of Giardia were found in faecal samples from 2 prehensile-tailed porcupines (Coendou villosus). The three positive animals for Cryptoporidium were rodents - 1 montane akodont (Akodon montensis), 1 ebony akodont (Thaptomyces nigrita) and 1 guainan squirrel (Sciurus aestuans). Microporidia spores were seen in the stools of 12 animals - 6 small rodents, including 3 montane akodonts, 1 prehensile-tailed porcupine and 2 pigmy rice rats (Oligoryzomys sp.); 3 marsupials, including 1 gray slender mouse opossum (Marmosops incanus) and 2 big eared opossums (Didelphis aurita); 3 hairy-legged vampire bats (Diphylla ecaudata). This is the first description of microsporidiosis in wildlife animals in Brazil. The present study emphasizes the importance of these animals, particularly small mammals, as potential sources of protozoan infection to other animal populations, including man, in areas of deforestation.

Experiencia terapéutica con nitazoxanida en diarrea crónica por microsporidios y criptosporidios en pacientes inmunocomprometidos oncológicos / Therapeutic experience with nitazoxanide in chronic diarrhea for microsporidia and cryptosporidia in immunocompromised cancer patients

Informar la experiencia de erradicación de Cryptosporidium y Microsporidium spp., en pacientes inmunocomprometidos con diarrea crónica utilizando nitazoxanida como terapia. Se evaluaron 50 pacientes con diarrea crónica (38 mujeres y 12 hombres), edad promedio de 45 y 56 años respectivamente; mayores de edad, dieron su consentimiento informado por escrito; con diagnósticos de cáncer de vías digestivas (veintidos), mama (diez y seis), cuello uterino (siete) y próstata (cinco); además 6 presentaban infección por VIH. Se hizo diagnóstico parasitológico empleando la coloración de Kinyoun. Recibieron nitazoxanida 500 mg/VO/BID por períodos de 3,6,9 días dependiendo del resultado de los controles microbiologicos. Se identificó como agente único a Cryptosporidium spp. en 2 pacientes y Microsporidium sp., en 6 pacientes, y combinados en 42 pacientes. Al tercer día de tratamiento, en 30 pacientes se logró erradicación de la criptosporidiosis, persistiendo con clínica diarreica en 28 de ellos; al sexto día se evidenció cura microbiológica (erradicación parasitaria de ambos microorganismos) y clínica (cese diarrea), en un total de 40 pacientes; al noveno día de tratamiento, se logró cura clínica y microbiológica en todos los pacientes incluyéndose en este último grupo todos los pacientes con infección VIH. Se evidenció epigastralgia en 4 pacientes que recibieron 9 días de terapia. Nitazoxanida parece ser un tratamieto eficaz y seguro en diarrea crónica micro y/o criptosporidiana en pacientes inmunocomprometidos oncológicos y es necesario realizar pesquisa microbiológica de parasitosis emergente y prolongación del tratamiento más allá de losa 3 días convencionalmente recomendados.

[Study of mortality agents of silk worms in Dough Abad]

For investigation of causes of silkworm mortality, the sick larva specimens in different levels of growth examined for parasite, viral, fungal and bacterial diseases. The samples included some dead larva and bed materials contained eggs, leaves and stool which were examined ordinary diagnostic methods. The complementary tests were also used for all cases. In wet smear prepared from intestine wall, a kind of protozoa spore of Microsporidia phylum was observed that was not the agent of pebrin disease, and some budding cells [yeasts] were seen as well, which identified as Saccharomyces cerevisiae. There were many polyhedra forms in smears of hemolynph that were stained with methylen blue dying and seen with ordinary microscope, diagnosed as Polyhedrosis or Grasserie disease [NPV]. for the purpose of diagnostic reconfirmation, the polyhedra was injected to healthy larva. The bacterial agent including Streptococcus [delta] Hemolytica as digestive tracts disease agent, staphylococcus aureus, and Serratia marcescen as septicemia agents and Bacillus turingensis the agent of Sotto disease, were separated. In general the incidence of these diseases showed the unsuitable conditions in sericultures that can be depends on poor disinfection, lack of proper feeding and deficiency of temperature and humidity. Improvement in rearing techniques, using of appropriate rearing room and equipments, maintenance of hygienic conditions during rearing, improvement of disinfection methods, the quarantine of the silkworm seeds and using of resistant varieties of silkworm to diseases can decrease out-breaks

Microsporidia intestinales en pacientes viviendo con SIDA en Honduras / Intestinal microsporidiosis in patients with AIDS in Honduras

ANTECEDENTES: Organismos pertenecientes al grupo Microspora son conocidos mundialmente como agentes de enfermedad oportunista en pacientes viviendo con SIDA. OBJETIVOS: Investigar la presencia de especies de microsporidia y la coinfección con parásitos intestinales en pacientes viviendo con SIDA en Honduras. METODOLOGÍA: Se examinó una muestra de heces de cada uno de 56 pacientes viviendo con SIDA del Hospital Mario Catarino Rivas, San PedroSula, Honduras, de mayo a agosto 1999, por la técnica de reacción en cadena de la polimerasa (PCR) y por coloraciones histoquímicas con Calcofluor Blanco 2MR y tricromo modificado y por cuatro métodos adicionales de rutina investigando la coinfección con parásitos intestinales. RESULTADOS: Se identificó esporas de microsporidia en 25 (14/56) de pacientes, por coloraciones histoquímicas de heces. La técnica de PCR, más sensitiva, identificó 41.5 (22/53) de pacientes con infección por microsporidia. Diez resultaron Enterocytozoon bieneusi o especies de Encephalitozoon; uno resultó E. bieneusi, 6 fueron positivos para Encephalitozoon spp, y 7 pertenecían a especies no determinadas. Diez y nueve de 22 individuos (86.3) identificados con esporas de microsporidia tenían también parásitos intestinales: Isospora belli (30.4), Strongyloides stercoralis (21.4), Ascaris lumbricoides(17.1), Cryptosporidium spp. (16.1), Trichuris trichiura (14.3) y uncinaria (10.7). Quistes de Giardia lamblia, Entamoeba histolytica/E. dispar,ooquistes de Cyclospora cayetanensis y huevos de Hymenolepis nana se encontraron en menor porcentaje.CONCLUSIÓN: Este estudio permitió identificar por primera vez infecciones por microsporidia intestinales en personas viviendo con SIDA en Honduras, solas o en asociación con otras infecciones por nemátodos,céstodos y protozoos comunes y/u oportunistas...

Influence of temperature on microsporidia infections in a natural population of Simulium pertinax Kollar, 1832 (Diptera; Simuliidae)

During the studies involving the correlation between the water temperature of the breeding site of Simulium pertinax larvae and the infection prevalence by microsporidia, developed in the Andorinhas river, Magé, RJ, weekly samples of blackfly larvae were taken within a two-year period (2001-2002 and 2003-2004), and it was noticed that the infections by Amblyospora sp. were more prevalent when compared to infections by Polydispyrenia sp. in larvae. It was also observed that the infections do not follow the same pattern, since the genus Amblyospora was recorded almost every month during the study with the exception of December, 2001. In the results of correlation between the environmental water temperature and the microsporidia infection rates, it was observed that for the first period studied, there was a high negative correlation, while during the second period there was absence correlation. On the other hand, the Amblyospora sp. infection rates prove that the correlation was high and significant in the first period, but was not significant in the second sampling period and Polydispyrenia sp. showed absence correlation in both periods.

Este estudo envolvendo a correlação entre a temperatura da água do criadouro de larvas de Simulium pertinax e a prevalência de infecção por microsporídeos, foi desenvolvido no rio Andorinhas, Magé, RJ, onde foram realizadas coletas semanais de larvas de simulídeos no período de dois anos (2001-2002 e 2003-2004), e foi observado que as infecções por Amblyospora sp. apresentaram maior prevalência quando comparadas com as infecções por Polydispyrenia sp. Com isso, verificou-se que as infecções não seguem o mesmo padrão, onde o gênero Amblyospora foi relatado em quase todos os meses de desenvolvimento do estudo, com exceção apenas de dezembro de 2001. Nos resultados de correlação entre a temperatura ambiente e a taxa de infecção por microsporídeos no primeiro período de estudo, foi observada uma forte correlação negativa, porém foi verificada ausência de correlação no segundo período. Embora tenha sido evidenciada uma forte e significante correlação com a taxa de infecção por Amblyospora sp. no primeiro período, esta correlação mostrou-se não significativa no segundo período de amostragem e Polydispyrenia sp. apresentou ausência de correlação em ambos os períodos de estudo.

Prevalence des infections opportunistes digestives parasitaires a Kinshasa (Republique Democratique du Congo). Resultats d'une enquete preliminaire chez 50 patients au stade SIDA

En Republique Democratique du Congo (RDC); comme dans de nombreux pays africains; le sida et son cortege d'infections opportunistes sont une cause majeure de morbidite et de mortalite. A Kinshasa; on estime entre 4 et 5le taux de prevalence de sujets infectes par le VIH; soit plus de 200 000 personnes (chiffres du Programme National de Lutte contre le Sida; PNLS 2005). A ce jour; faute de personnels formes et de moyens diagnostiques adaptes; aucune enquete n'a encore ete menee sur la prevalence des parasites opportunistes digestifs dans la population des patients infectes par leVIH; prealable indispensable a la mise en place d'une politique de soin adaptee. Une enquete preliminaire a ete realisee a Paris sur 50 echantillons de selles de 50 patients malades du sida; hospitalises dans 3 hopitaux de references de Kinshasa. Onze patients (22) avaient une symptomatologie digestive avec un syndrome diarrheique. La realisation des examens specialises a mis en evidence 2 cas d'infection digestive par des parasitoses opportunistes (4); une a Cryptosporidium sp. et une a Enterocytozoon bieneusi; premier cas decrit dans la litterature en RDC

Heterosporis anguillarum infections in farm cultured eels (Anguilla japonica) in Korea

Ten eels (Anguilla japonica) from a fish farm in Korea were examined and diagnosed with a Heterosporis infection. The gross lesions on the trunk were uneven and the concave parts were pasty. Histopathologically, lyses of the trunk muscles, degenerative muscle fibers and the scattered spores were observed. The sporophorocyst (SPC) contained several spores with a variety of shapes. Some SPC were disrupted and the spores in the SPC were scattered in the muscle tissues. Macrophages existed near the scattered spores. Electron microscopy revealed special structures such as sporophorocyst containing various developmental parasitic stages such as meronts, sporonts, sporophorous vesicles and spores.

Microsporidiosis in a Brazilian University Hospital: case report

This is the report on a patient with chronic diarrhea caused by microsporidia. He is married, infected with HIV and has low CD4 cell count. The diagnosis was established through stool parasite search using concentration methods and Gram - chromotrope staining technique. Ileum biopsy was also performed in this case. The etiological diagnosis may be established in a clinical laboratory, by chromotrope staining technique in routine microscopic examination of stool specimens.

Este é o relato de caso de doente com diarréia crônica causada por Microsporidia. O doente era homem, casado, infectado com HIV e tinha baixa taxa de linfócitos CD4+. O diagnóstico foi feito em exame de fezes utilizando métodos de concentração e técnica de coloração de Gram-Chromotrope. Biópsia de íleo também foi realizada neste caso. O diagnóstico etiológico pode ser feito em laboratório clínico, por técnicas de coloração baseada em cromotrope na rotina da observação microscópica direta.

Observations on the intracytoplasmic microsporidian Steinhausia mytilovum, a parasite of mussel (Mytella guyanensis) occytes from the Amazon river estuary

Microsporidians (Microsporidia) can parasitize commercially important marine mollusks, including bivalves. In this report, we provide a brief description of the ultrastructure of the microsporidian Steinhausia mytilovum that occurs in the oocyte cytoplasm of the mussel Mytella guyanensis (Mollusca, Bivalvia, Mytillidae) from the Amazon river estuary. Mussel ovaries were fixed, stained and examined using differential interference contrast optics (DIC). The parasite developed in an intracytoplasmic vacuole containing a variable number of spores (up to 14). Mature spores were 2.3 ± 0.3 ìm long and 1.7 ± 0.3 ìm wide (n = 25 each). Transmission electron microscopy revealed two types of intracytoplasmic vacuoles, one containing spores with a light (less dense) cytoplasm that corresponded to the maturation phases, and the other containing mature, dense, granular spores that showed specific microsporidian structures. The anchoring disc and the anterior zone of the polar filament were surrounded by the polaroplast. The polar filament was isofilar and consisted of a double (rarely triple) coil with 9-10 turns. The ultrastructural morphology of these spores suggested that they belonged to S. mytilovum.