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PubMed2003

Lectin histochemistry for glycoconjugates in the small intestines of piglets naturally infected with Isospora suis.

Composition of glycoconjugates was examined in small intestines naturally infected with Isospora suis in preweaned pigs by use of 21 biotinylated-labeled lectins with avidin-biotin-peroxidase complex method. As compared with control pig, staining of 18 lectins altered in jejunal villus brush border and goblet cells of pigs naturally infected with I. suis. These results indicate that I. suis infection alters carbohydrate residues on the jejunal intestines.

Animals↗
PubMed2004

Point prevalence of Cryptosporidium, Cyclospora, and Isospora infections in patients being evaluated for diarrhea.

From March to September 2001, 315 specimens from "nonrepeat" patients that were submitted for ova and parasite examination were stained using the Kinyoun modified acid-fast stain to detect the intestinal coccidians. Four patients (1.3%) were infected with coccidians, 2 with Cryptosporidium parvum and 2 with Cyclospora cayetanensis. No infections with Isospora belli were detected. In comparison, 15 patients (4.8%) had infections with one or more intestinal parasites detected by routine trichrome staining: 5 had Giardia lamblia; 2, Dientamoeba fragilis; 3, Strongyloides stercoralis; 1, Iodamoeba bütschlii; 3, Endolimax nana; 6, Blastocystis hominis; and 1, Entamoeba coli. Four patients were multiply infected. Coccidians made up 29% of the clinically significant parasitic infections. The coccidians were missed in all 4 cases because no special staining was ordered. Clinicians need to be reminded that additional tests should be ordered to fully evaluate patients with chronic diarrhea in which no diagnosis is found by routine testing.

Adolescent↗
PubMedDate not supplied

[Comparison between the modified Ziehl-Neelsen and Acid-Fast-Trichrome methods for fecal screening of Cryptosporidium parvum and Isospora belli].

Due to the importance of Cryptosporidium parvum, Cyclospora cayetanensis and Isospora belli as opportunistic parasites, it is essential for laboratories to morphologically differentiate these coccidia. Among the recommended methods for diagnosis of C. parvum and I. belli is the Modified Ziehl-Neelsen stain (MZN) and recently a new technique, the Acid-Fast-Trichrome (AFT) was proposed. The purpose of this study was to compare the AFT and MZN techniques regarding the detection of C. parvum and I. belli oocysts in stool samples of HIV-positive patients. According to the presence (n=60) or absence of diarrhea (n=60), two groups were selected for inclusion in this study. The stool samples were collected in 10% formalin solution and the pellets, obtained without and after centrifugation-concentration (500xg; 10 min), were submitted to both stain techniques. Considering the positivity of the techniques (AFT and MZN), we observed the superiority of MZN stain (n=19; 100% positive cases) over AFT (n=8; 42.1%). The identification of the 101 truly-negative cases was possible with both techniques, but when the AFT stain was used, the oocysts showed a variable inner stain degree, which resulted in false-negatives. Enteric coccidiosis was more frequent among the patients who presented diarrhea (26.6%) compared to asymptomatic individuals (5%); C. cayetanensis was not detected in these groups. We were interested in evaluating the employment of AFT for the Cyclospora oocyst stain. Based on the sensitivity and specificity obtained in this study (100%), we concluded that the MZN technique is still the most indicated for routine use in clinical analysis laboratories, for the diagnosis of cryptosporidiosis and isosporosis, especially when it was used together with the centrifugation procedure (500xg; 10 min). Our results showed that with some improvements, AFT would be a simple and inexpensive technique appropriate for use in the diagnosis of intestinal coccidia (C. parvum, C. cayetanensis and I. belli). It can also be extended to the detection of microsporidia.

Animals↗
PubMed2003

[Extraintestinal finding of Isospora belli unizoic cysts in a patient with AIDS: case report].

This report describes the presence of Isospora belli unizoic cysts in mesenteric lymph nodes and of gametocytes in the gallbladder epitelium of a 26 year-old Brazilian male patient with Acquired Immune Deficiency Syndrome. This patient had received treatment for several times with sulfamethoxazole-trimethoprim. It is discussed the significance of I. belli tissue cysts as possible foci of resistance of the parasite and their association with the infection relapse even post-treatment with anticoccidian medication.

AIDS-Related Opportunistic Infections↗
PubMedDate not supplied

Experimental infection of swine by Isospora suis Biester 1934 for species confirmation.

A survey of Isospora suis performed in 177 faecal samples from 30 swine farms detected thin wall type I. suis oocysts in seven samples. This type of oocyst measuring 23.9 by 20.7 microns had a retracted thin wall similar to that of the genus Sarcocystis. This type of oocysts, isolated from four different faecal samples, was inoculated in four-five-days-old piglets free of contamination in order to verify the life cycle and pathogenicity of the species. The pigs were kept in individual metal cages and fed with cow milk. Daily faecal collections and examinations were performed until the 21st day after infection. MacMaster and Sheather's methods were used for oocyst counting and identification. Infected piglets produced yellowish-pasty diarrhoea with slight dehydration. The prepatent and patent periods were respectively from 6 to 9 and 3 to 10 days after infection. Oocyst elimination was interrupted on the 10th and 11th days after infection with biphasic cycles. Thin and thick wall oocysts were detected in the same faecal samples. Thin walls were not observed in unsporulated oocytes. The observations suggest that this type of oocysts could appear in specific strains which occur in the later stages of their development. These oocysts seem to be responsible for clinical and pathogenic signs of neonatal isosporosis in pigs.

Animals↗
PubMedDate not supplied

The fine structure of the endogenous stages of Isospora hemidactyli Carini, 1936 in the Gecko Hemidactylus mabouia from North Brazil.

The ultrastructure is described of the meronts, microgamonts and young oocyst stages of Isospora hemidactyli of the gecko Hemidactylus mabouia from Belém, PA, north Brazil. The endogenous stages all develop in the nucleus of the gut epithelial cells. The nucleus remains intact up to the latest stages of the parasite's development, but degenerates by the time the oocyst appears. Merogonic division appears to be asynchronous, and some of the differentiated merozoites contained more than one nucleus. Microgamonts conform in structure with those of other eimeriids. Some of the type 2 wall-forming bodies disintegrate into smaller globules and ground substance of lower density.

Animals↗
PubMed2002

Clinical large intestinal coccidiosis in camels (Camelus dromedarius) in the United Arab Emirates: description of lesions, endogenous stages, and redescription of Isospora orlovi, Tsygankov, 1950 oocysts.

Between January and March 2001, eight 4- to 8-wk-old camels (Camelus dromedarius) from 2 farms from Dubai area of the United Arab Emirates were submitted for necropsy examination. The camels had diarrhea of 2-5 days duration. Grossly, a severe diphtheroid-to-hemorrhagic colitis was seen in all animals. Gamonts, unsporulated oocysts, sporulating oocysts, and fully sporulated oocysts were present in the intestinal epithelium and the lamina propria. Fully sporulated oocysts contained 2 sporocysts and 4 sporozoites in each sporocyst. Oocysts from fecal samples resembled oocysts of Isospora orlovi. This is the first report of an isosporan parasite associated with hemorrhagic enteritis in the large intestine of any animal.

Animals↗
PubMed2005

Molecular characterization of isosporoid coccidia (Isospora and Atoxoplasma spp.) in passerine birds.

Prevalence and disease caused by isosporoid coccidia in passerine birds are well recognized, but confusion about the life cycles of the parasites has led to taxonomic inconsistencies. In this study, we characterized segments of the chromosomal small and large-subunit ribosomal RNA (rRNA) genes of coccidial parasites from 23 species of passerine birds, as well as heat shock protein 70, apicoplast rRNA, and chromosomal 5.8s rRNA genes from a subgroup of these animals, and we correlated genetic data with morphologic findings for different parasite developmental stages, host phylogeny, and overall taxonomic relations within the phylum Apicomplexa. Our findings indicate that isosporoid coccidia of passerine birds are monophyletic but exhibit substantial diversity, with most avian species having one or several unique parasite lineages that underwent synchronous speciation with their hosts, interrupted by sporadic episodes of lateral transmission across species and families. Molecular analyses support a homoxenous life cycle, with sexual forms occurring chiefly in the intestines and asexual merozoites present systemically. Rarely, extraintestinal sexual stages can occur. The passerine coccidia are genetically most closely related to species of Eimeria rather than Isospora. We suggest that these parasites, whether identified from blood merozoite stages or fecal oocysts, be provisionally grouped as a homogeneous clade of individual species in a single taxon and formally named when reliable criteria allowing reclassification of related genera in the suborder Eimeriina are clarified.

Animals↗
PubMedDate not supplied

Submicroscopic profile of Isospora belli enteritis in a patient with acquired immune deficiency syndrome.

Small bowel mucosal fragments from a human immunodeficiency virus-positive female patient with chronic diarrhea were investigated by transmission electron microscopy, and Isospora belli enteritis was documented. The submicroscopic profile was characterized by a moderate abnormality of mucosal architecture with reduction in height of villi and hypertrophy of crypts. Stages of both asexual (trophozoite, schizont and merozoite) and sexual (macrogametocyte) phases of the life cycle of the parasite were identified in the epithelium, always enclosed within a parasitophorous vacuole. Moreover, the presence of occasional extracellular merozoites in the intestinal lumen and in the lamina propria near or within lymphatic vessels was documented. These findings expand the current knowledge of this parasite regarding its capacity to survive in an extracellular environment and document a possible mechanism by which extraintestinal infection can take place.

AIDS-Related Opportunistic Infections↗
PubMed1978

"Ghost" forms of Giardia lamblia cysts initially misdiagnosed as Isospora.

Because of their distinctive morphology the cysts of Giardia lamblia usually are not mistaken for cysts or eggs of other intestinal parasites. At a hospital laboratory in Kentucky a case of giardiasis was encountered in which the initial fecal examination revealed only degenerated cysts of Giardia which were mistaken for the oocysts of Isospora at the one-sporoblast stage.

Adult↗
PubMed2000

Trimethoprim-sulfamethoxazole compared with ciprofloxacin for treatment and prophylaxis of Isospora belli and Cyclospora cayetanensis infection in HIV-infected patients. A randomized, controlled trial.

BACKGROUND: In developing countries, Isospora belli and Cyclospora cayetanensis frequently cause chronic diarrhea in HIV-infected patients. OBJECTIVE: To compare 1 week of trimethoprim-sulfamethoxazole treatment and 1 week of ciprofloxacin treatment in HIV-infected patients with chronic diarrhea caused by I. belli and C. cayetanensis. DESIGN: Randomized, controlled trial. SETTING: HIV clinic in Port-au-Prince, Haiti. PATIENTS: 42 HIV-infected patients with chronic diarrhea due to I. belli (n = 22) or C cayetanensis (n = 20). INTERVENTIONS: Patients were randomly assigned to receive oral trimethoprim-sulfamethoxazole (160 mg or 800 mg) or ciprofloxacin (500 mg) twice daily for 7 days. Patients who responded clinically and microbiologically received prophylaxis for 10 weeks (1 tablet orally, three times per week). MEASUREMENTS: Treatment success was measured by cessation of diarrhea and negative stool examination at day 7. Prophylaxis success was measured by recurrent disease rate. RESULTS: Diarrhea ceased in all 19 patients treated with trimethoprim-sulfamethoxazole. Eighteen of 19 patients had negative results on stool examination at day 7 (95%). Among the 23 patients who received ciprofloxacin, diarrhea ceased in 20 (87% [CI; 66% to 97%]) and 16 had negative results on stool examination at day 7 (70%). By survival analysis, diarrhea from isosporiasis and cyclosporiasis ceased more rapidly with trimethoprim-sulfamethoxazole than with ciprofloxacin. All patients receiving secondary prophylaxis with trimethoprim-sulfamethoxazole remained disease-free, and 15 of 16 patients receiving secondary prophylaxis with ciprofloxacin remained disease-free. CONCLUSIONS: A 1-week course of trimethoprim-sulfamethoxazole is effective in HIV-infected patients with cyclosporiasis or isosporiasis. Although ciprofloxacin is not as effective, it is acceptable for patients who cannot tolerate trimethoprim-sulfamethoxazole.

AIDS-Related Opportunistic Infections↗
PubMedDate not supplied

[A new trichromic safranin stain for the detection of Cryptosporidium parvum, Cyclospora cayetanensis, species of Microsporidia and Isospora belli in fecal material].

Cryptosporidium parvum, Isospora belli, Cyclospora cayetanensis and Microsporidia are frequent pathogens in the immunodeficient host, which may cause multiple infections. The above mentioned parasites are found in feces by the application of different specific tintorial techniques. The objective of this work was the development of a stain for the simultaneous detection of these parasites, reducing costs as well as the time taken to make the diagnosis. The safranin-trichrome stain is simple, chip and its results are similar to those of specific tints. All microorganisms are easy to detect and besides being perfectly distinguishable from fungi and faecal elements.

AIDS-Related Opportunistic Infections↗
PubMed1996

Uvitex 2B stain for the diagnosis of Isospora belli infections in patients with the acquired immunodeficiency syndrome.

OBJECTIVE: Fluorescent stains with Uvitex 2B or other fluorochromes are widely used today, especially for the diagnosis of microsporidian infections in human immunodeficiency virus (HIV)-infected patients. Our objective was to ascertain whether the fluorescent stain with Uvitex 2B is also able to detect Isospora belli in stool samples and duodenal/bile juice of HIV-infected patients. DESIGN: Case study. SETTING: University hospital of the University of Cologne, Germany. PATIENTS: Two HIV-infected patients with chronic diarrhea who had intestinal infections with I. belli. METHODS: Stool was concentrated by a modified water-ether sedimentation method, and duodenal/bile juice was concentrated by centrifugation. Wet-mount preparations were examined by phase-contrast and bright-field microscopy, and smears were stained with a modified acid-fast stain and a fluorescent stain with Uvitex 2B. RESULTS: Using the fluorescent stain with Uvitex 2B, the oocysts of I. belli stained bright white/blue fluorescent and showed a structure similar to that of the oocysts in acidfast stains. CONCLUSIONS: Staining with Uvitex 2B is a suitable method for the diagnosis of I. belli infections. This technique can be used for the diagnosis of three important gastrointestinal parasites (viz, microsporidia, cryptosporidia, and I. belli) responsible for diarrhea in HIV-infected patients.

AIDS-Related Opportunistic Infections↗
PubMed1992

Isospora solimanae sp. nov. from the Egyptian hoopoe (Upupa epops major).

Isospora solimanae (sp. nov.) is recorded for the first time infecting the Egyptian hoopoe (Upupa epops major). Unsporulated as well as sporulated oocysts were studied, photographed and compared with other isosporan infections of related birds. Site of infection and invasion limits were determined among naturally infected hoopoes and represented diagramatically. The rate of infection was congruent to 37.5%, and its severity considered to be non-pathogenic, since no remarkable lesions were observed. No extra-intestinal site of parasite development was recorded.

Animals↗
PubMed2003

Cryptosporidium parvum and Isospora belli infections among patients with and without diarrhoea.

OBJECTIVE: To assess the importance of Cryptosporidium parvum and Isospora belli infections as a cause of diarrhoea among patients admitted to the Medical Wards in Queen Elizabeth Central Hospital (QECH) in Blantyre, Malawi. DESIGN: Prospective case control study. SUBJECTS: One hundred and twenty one patients with diarrhoea and 122 patients without diarrhoea. MAIN OUTCOME MEASURES: Demonstration of C. parvum and I. belli oocysts by examination of at least one stool sample per patient using phenol auramine-O-fluorescence staining and an immuno-fluorescent assay with monoclonal antibodies against Cryptosporidium, seropositivity for HIV and AIDS. RESULTS: In 22% of the patients with diarrhoea an infection with C. parvum or I. belli was found. Thirteen (11%) of them had a C. parvum and 14 (12%) an I. belli infection; a mixed infection was found in one patient. In the control group, three (3%) C. parvum and three (3%) I. belli infections were seen. The prevalence of both infections was very significantly higher in the cohort of diarrhoea patients than in the controls, 13/108 versus 3/119 (p=0.0099) for C. parvum, and 14/107 versus 3/119 (p=0.0056) for I. belli. Infections were only seen in HIV positive patients. Two hundred and four (84%) patients were HIV positive and 145 (60%) of them had AIDS. CONCLUSIONS: C. parvum and I. belli infections are a significant cause of diarrhoea among medical in-patients at QECH. Examinations of stool specimen for parasites among hospitalised patients with diarrhoea provide data for a more appropriate management.

AIDS-Related Opportunistic Infections↗
PubMed2004

Development of Isospora belli in Hct-8, Hep-2, human fibroblast, BEK and Vero culture cells.

The development of Isospora belli, a human coccidian parasite, was studied in different cell lines. Merozoites were observed in all kinds of cells, whereas sporogony was demonstrated only in Hct-8. This implied that not only the human cell line can be infected, but also some animal cell lines. Unizoites could be found in Vero cells. The merozoites were transferred to a new culture cell for three passages and maintained for two weeks, but no oocyst production was observed in any culture cells during cultivation.

Animals↗
PubMedDate not supplied

[Comparison of different methods for examining the feces of suckling piglets for Isospora suis].

A highly sensitive diagnostic method is of great importance for detection of I. suis (Isospora suis). In the present study KSFV (combined sedimentation-flotation method) using different flotation solutions (NaCl, ZnSO4, NaCl-sucrose) was investigated regarding sensitivity, costs and expenditure of human labour in comparison to FV (flotation method) using NaCl-sucrose solution and to AM (autofluorescence microscopy). Ninety faecal samples were examined. Using AM, oocysts were detected in 83.3% of the samples. In cases of KSFV and FV, respectively, 41.1% to 53.3% of samples were tested positive. The quantity of detected oocysts was markedly higher in AM than in KSFV or FV. Therefore AM is the most sensitive of the comprised methods. The expenditure of labour is lowest in AM, time exposure is lower in FV than in KSFV. For performance of AM a fluorescence microscope is required contrary to KSFV and FV.

Animals↗
PubMed1991

[Lysosomal activity in enterocytes in the small intestine in piglets experimentally infected with Isospora suis].

The lysosomal activity of enterocytes of the small intestine mucosa was investigated in gnotobiotic and conventional piglets experimentally infected on the first day after birth (DAB) by the oocysts of the coccidia Isospora suis. A method of the proof of beta-D-glucuronidase (EC.3.2.1.31.) activity was used to demonstrate lysosomes. The piglets were infected by different infection doses of oocysts (100,000 oocysts in gnotobiotic piglets and 200,000 oocysts in conventional piglets). In the gnotobiotic infected piglets the activity of beta-D-glucuronidase in enterocyte lysosomes was investigated in the period from day 3 to day 11 after infection (DAI) and in the infected conventional piglets in the period from day 2 to day 10 after infection. Comparing the control piglets, the group of gnotobiotic piglets at the age of 2-5 days and the group of conventional piglets at the age of 4-7 days, the higher activity of beta-D-glucuronidase was demonstrated in the lysosomes of intestinal mucosa enterocytes in the gnotobiotic control piglets (+5.30 of the average density value, Dx). In the infected gnotobiotic and conventional piglets the pattern of beta-D-glucuronidase activity was found to have three stages in the course of this infection. Two stages can be characterized by a great increase in the enzyme activity (DAI 3-9 in gnotobiotic piglets, DAI 2-3 and 7-9 in conventional piglets. The third stage, which is manifest mainly in the conventional infected piglets, is characterized by a marked decrease in the activity of beta-D-glucuronidase, reaching the level of control findings (DAI 10 and mainly 11 in gnotobiotic piglets. DAI 4-6 and 10 in conventional piglets). A topographical picture shows that the two stages of increase and the stage of beta-D-glucuronidase activity decrease occur in the whole small intestine without any predisposition defect of the enzyme in the different sections of the small intestine.

Animals↗