Microsporidia are obligate intracellular parasites that infect a broad range of vertebrates and invertebrates. They have been increasingly recognized as human pathogens in AIDS patients, mainly associated with a life-threatening chronic diarrhea and systemic disease. However, to date the global epidemiology of human microsporidiosis is poorly understood, and recent data suggest that the incidence of these pathogens is much higher than previously reported and may represent a neglected etiological agent of more common diseases indeed in immunocompetent individuals. To contribute to the knowledge of microsporidia molecular epidemiology in HIV-positive patients in Nigeria, the authors tested stool samples proceeding from patients with and without diarrhea.
Stool samples from 193 HIV-positive patients with and without diarrhea (67 and 126 respectively) from Lagos (Nigeria) were investigated for the presence of microsporidia and Cryptosporidium using Weber’s Chromotrope-based stain, Kinyoun stain, IFAT and PCR. The Weber stain showed 45 fecal samples (23.3%) with characteristic microsporidia spores, and a significant association of microsporidia with diarrhea was observed (O.R. = 18.2; CI: 95%). A similar result was obtained using Kinyoun stain, showing 44 (31,8%) positive samples with structures morphologically compatible with Cryptosporidium sp, 14 (31.8%) of them with infection mixed with microsporidia. The characterization of microsporidia species by IFAT and PCR allowed identification of Enterocytozoon bieneusi, Encephalitozoon intestinalis and E. cuniculi in 5, 2 and 1 samples respectively. The partial sequencing of the ITS region of the rRNA genes showed that the three isolates of E.bieneusi studied are included in Group I, one of which bears the genotype B.
To our knowledge, this is the first report of microsporidia characterization in fecal samples from HIV-positive patients from Lagos, Nigeria. These results focus attention on the need to include microsporidial diagnosis in the management of HIV/AIDS infection in Nigeria, at the very least when other more common pathogens have not been detected.
Citation: Ojuromi OT, Izquierdo F, Fenoy S, Fagbenro-Beyioku A, Oyibo W, Akanmu A, et al. (2012) Identification and Characterization of Microsporidia from Fecal Samples of HIV-Positive Patients from Lagos, Nigeria. PLoS ONE 7(4): e35239. https://doi.org/10.1371/journal.pone.0035239
Editor: Elizabeth Didier, Tulane University School of Public Health and Tropical Medicine, United States of America
Received: January 13, 2012; Accepted: March 10, 2012; Published: April 9, 2012
Copyright: © 2012 Ojuromi et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This work was supported by a grant from Fundación San Pablo-CEU USP-PC03/08, and Lagos State University, Ojo, Nigeria, assisted for the visit of OTO to Spain. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
Diarrhea and severe weight loss are syndromes described in HIV patients and known as “slim disease” by sub-Saharan Africans . This pathology can be produced by several organisms, and it is commonly associated with Cryptosporidium. Additionally, microsporidia are considered as emerging pathogens worldwide and several species are involved in human disease . Among them, Enterocytozoon bieneusi and Encephalitozoon species, E. intestinalis, E. hellem and E. cuniculi are the most frequently identified microsporidia in human clinical samples. They have been described as opportunistic pathogens in human immunodeficiency virus (HIV)-infected patients and other immunocompromised patients such as transplant recipients , , , , , , , . However, microsporidia have also been detected in HIV-negative individuals , , , ,  and it has been recently suggested that the incidence of miscrosporidial infections in healthy population is much higher than previously reported and microsporidia may represent neglected etiological agents of more common diseases  In HIV-infected patients, especially those with CD4+ T-cell counts below 100 cells per mm3, E. bieneusi and E. intestinalis have been associated with acute and chronic diarrhea , , , , , , .
In the last decade, epidemiological studies of human microsporidiosis, have been strengthened with the improvement of diagnostic methods and the development of molecular markers . However, the transmission and sources of infection are not well understood. The transmission may involve person-to-person as well as waterborne or food contamination, especially in developing countries with poor sanitation . In the last few years several authors have focused on animal microsporidiosis, in order to elucidate the possible zoonotic origin of human microsporidiosis. In fact, human microsporidia species have been isolated from a large number of domestic and wild animals , , , . This zoonotic transmission is supported by phylogenetic studies which demonstrate that several genotypes can infect both humans and animals , , , .
The introduction of highly active antiretroviral therapy (HAART) with protease inhibitors to treat HIV/AIDS patients have substantially decreased the incidence of microsporidiosis in Europe . However the situation differs in developing countries, where the rapid expansion of AIDS together with limited access to HAART has contributed to an increased incidence of this disease. In the studies carried out in Africa to evaluate the prevalence of microsporidiosis in HIV–infected patients due to E. bieneusi and Encephalitozoon species, a variable incidence ranging from 7 to 51% was obtained, depending on the population studied and the diagnostic methods used , , , , , , , , . Unfortunately, epidemiological studies of microsporidiosis based in genotyping methods are scarce in these countries , , , , , so there is still insufficient information on the magnitude of the microsporidial infection in African countries. The most likely explanations are the lack of an easy to make diagnostic method specific for the detection of the parasite and the lack of experienced and skilled microscopists able to detect the organism in biological samples.
With the aim of contributing to the knowledge of the molecular epidemiology of microsporidia in Nigeria, we performed a study in HIV-infected patients with and without diarrhea. Additionally, and simultaneously, the presence of Cryptosporidium in these patients was also determined.
Materials and Methods
The protocol and studies were approved by the Institutional Review Board of the College of Medicine of the University of Lagos, Nigeria (CMUL) and the Nigerian Institute of Medical Research (NIMR), Yaba, Lagos, Nigeria. All participants gave informed consent before their samples were collected and processed.
A total of 193 patients were enrolled in the study, all of them were HIV–infected patients attending to referral clinics in Lagos, Nigeria, 67 (34.7%) with and 126 (65.3%) without diarrhea. Chronic diarrhea was either continuous, defined by the Center for Disease Control and Prevention as two or more loose stools per day for more than 28 days; or acute, defined as episodes of two or more loose stools per day alternating with episodes of formed stools for less than 28 days. Complete medical histories were obtained from the hospital records (Demographic and Clinical) for patients included in the study.
Stool Sample Collection, Storage and Staining Methods
Fresh stool samples were collected in clean universal bottles labeled with each patient’s details. Samples were stored at -80°C until use. Thin smears were made from all fecal samples to be stained with Webeŕs Cromothrope stain and Kinyoun stain to detect the presence of microsporidia and tentatively Cryptosporidium sp , .
Indirect Immunofluorescence Antibody Test (IFAT)
This was performed on all samples which tested positive by Webeŕs Cromothrope stain. IFAT was performed by coating the slides with 5 µl of each unconcentrated stool sample. Specific polyclonal antibodies from rabbit for E. hellem (1/500), E. intestinalis and E. cuniculi (1/400) , ,  and a monoclonal antibody for E. bieneusi  were used. Subsequently, the fluorescein isothiocyanate-conjugated anti-rabbit (Sigma Cat. F-9887) and anti-mouse (Sigma Cat. F-1010) were appropriately diluted and the slides were examined with a fluorescence microscope.
DNA from fecal samples found to be positive for microsporidia by the staining method was extracted by bead disruption of spores using the Fast-DNA-Spin kit, according to the manufactureŕs instructions (Bio 101, Carlsbad, Calif.). PCR inhibitors were removed using the QIAquick PCR kit (QIAGEN, Chatsworth, CA).
Species characterization by PCR.
Microsporidial small subunit rRNA (SSU-rRNA) coding regions were amplified using the following species-specific primers EBIEF1/EBIER1 for E. bieneusi , SINTF/SINTR for E. intestinalis , ECUNF/ECUNR for E. cuniculi and EHELF/EHELR for E. hellem . PCR amplification was performed with the GeneAmp kit (Perkin-Helmer Cetus, Norwalk, CT) according to the manufacturer instructions. The conditions for the reaction and the testing of purified samples for the presence of PCR inhibitors by spiking the samples with the corresponding cloned SSU-rRNA coding region was as described .
Genotyping by DNA sequencing analysis.
E. bieneusi genotypes from 3 fecal samples were analyzed by nucleotide sequencing of the Internal Transcribed Spacer (ITS) of the rRNA genes. The PCR products were purified using Concerted Rapid PCR kit (GIBCO-BRL) and sequenced in both directions using an ABI Big Dye Terminator kit (v1.1) and an ABI 3100 automated sequencer (Applied Biosysytems). The sequences were edited with BIOEDIT 7.0 program  and aligned following the Clustal W algorithm .
These were performed by comparison with 179 sequences available in the GenBank obtained from different hosts and reviewed by Henriques-Gil et al 2010 . The haplotypes were generated with the program DNASP 4.10 , without considering the occasional gaps. Haplotype Median Joining networks  were performed with NETWORK 22.214.171.124 program (Fluxus Technology Ltd.). The network was simplified reducing star-like clusters to the ancestral haplotypes with a single run of star contraction .
Forty-five stool samples (23.3%) showed a variable number of spores that stained pinkish red with the use of Weber’s Chromotrope, showing the typical characteristics of microsporidia (Table 1). These spores measured 1 to 2 µm and were ovoid in shape with a clear vacuole-like polar zone. From the 45 stool samples microsporidia positive by stain, 37 (52.2%) were obtained from patients with diarrhea and 8 (6.3%) from non-diarrheic samples (Table 1). The presence of microsporidia spores in fecal samples from patients with diarrhea was compared with that from patients with no diarrhea, revealing a clear association between microsporidia and diarrhea with O.R. = 18.2 (χ2 = 58.44; p< 10–6).
Structures morfologically compatible with Cryptosporidium sp were detected in 44 of the HIV patients (22.7%) using Kinyoun stain. Thirty-two of positive patients presented diarrhea and 12 were found in non-diarrheic samples, supporting the well-known influence of Cryptosporidium sp on diarrhea (χ2 = 36,34; p< 10–6 ; OR = 8,7) (Table 1). Mixed infection was recorded between microsporidia and Cryptosporidium sp in 14 patients (31.8% of positives) (Table 1). In patients with diarrhea, the two types of microorganisms could be considered antagonistic as mixed-infections were lower than expected (χ2 = 10.77; p< 0.01) (Table 1).
The study by IFAT of microsporidia positive samples showed 8 positive samples. Five samples exhibited bright fluorescence when E. bieneusi monoclonal antibody was used; two reacted with E. intestinalis polyclonal antibody and one with E. cuniculi polyclonal antibody. Spores which reacted with the anti-E. cuniculi and anti-E. intestinalis polyclonal antibodies were observed in non-diarrheic samples, and E. bieneusi spores were identified in 1 non-diarrheic and 4 diarrheic samples by the monoclonal antibody used (Table 2).
A total of 7 cases, five for E. bieneusi and 2 for E. intestinalis were confirmed by amplification of DNA isolated from positive samples in the staining technique with specific primers for the most common microsporidia infecting humans (Table 2).
DNA Sequencing Analysis
Three PCR positive samples for E. bieneusi yielded enough PCR products for partial sequencing of the ITS region of the rDNA genes. The Nigerian genotypes reported here are included in Group I. For comparison with 172 sequences from the GenBank, the gaps were excluded, and 139 sites were available for analysis. Fifty sites were polymorphic and generated 60 different haplotypes. One of the sequences obtained from the Nigerian isolates showed the same haplotype as genotype B . Since we were unable to analyze the entire ITS, the sequences of the other two isolates coincided with several similar genotypes: P, type IV, UG2145, Peru3, PtEb IV or PtEbV . The diversity in the ITS of E. bieneusi is very high but, within Group I, the differences between any pair of sequences involve only a small number of mutational events.
In order to locate the Nigerian samples within the wide variability of E. bieneusi, Median Joining networks were constructed with the program NETWORK . The network was simplified, reducing star-like clusters to the ancestral haplotypes with a single run of star contraction . The network with the resulting 40 taxa is shown in Figure 1. Nigerian haplotypes, together with the other African isolates previously analyzed, occupy a central position in the network.
Circles are proportional to the frequency of each haplotype. The colour of each sector refers to the host species: Nigerian samples (squared sectors), other African haplotypes (black), Europe and Asia (grey lines), North and South America (white). The lengths of the segments are proportional to the number of mutations involved between two haplotypes.
Diarrhea caused by opportunistic agents such as microsporidia, Cryptosporidium species or Isospora belli contributes to morbidity and mortality in AIDS patients. In developing countries, these infections taken together may likely account for a great number of cases of diarrhea in HIV/AIDS patients . To date there is only two reports included in PubMed of microsporidia infection in patients from Nigeria. In the study carried out in a rural village (Magama Gumau) and in a township in north central Nigeria a prevalence of 39.6% and 47.3% in rural and urban dwellers was reported . However, the stain method used to detect microsporidia was not Webeŕs reference method or Gram-chromotrope stain and no characterization of the microsporidia species was made. More recently, a study was carried out in children attending an outpatient department of a hospital in Oyo state, southwestern Nigeria . Using PCR, 9.3% of the children were identified as positive for E.bieneusi. Therefore, to contribute to a more complete knowledge of microsporidia epidemiology in Nigeria, this study has evaluated and characterized the presence of microsporidia in 193 fecal samples from HIV-positive patients with and without diarrhea (67 and 126 samples respectively); showing 45 positive samples (23.3%) when Webeŕs stain was used. Most of the positive samples were identified in diarrheic stools (52.2%) and only 8 in non-diarrheic stools (6.3%), showing a significant association between microsporidia and diarrhea in the population studied (OR = 18.2) such as had previously been reported in other countries , , , , . Our results are in accordance with previous studies on the prevalence of microsporidia in HIV-positive patients, which ranges from 5 to 50% depending on the geographical location and the diagnostic technique employed . This suggests that microsporidia are becoming highly prevalent intestinal parasites in developing countries such as Nigeria and thus the need to consider them as neglected etiological agents . In reference to the African continent, our results are also in accordance with previous studies in HIV-positive patients with chronic diarrhea , .
To determine the species of microsporidia implicated in the study, two additional methods were used: IFAT and PCR (the gold standard). The use of IFAT allowed the identification of microsporidia species in only 8 fecal samples (Table 2).We detected a low positive result by IFAT compared with Webeŕs staining. However, the scarce parasitic load and the low specificity of Weber’s staining could be related with the differences observed. However, we can not discard the possibility that other microsporidia species were present in our samples.
In our study most of the positive samples by IFAT were E. bieneusi (4) followed by E. intestinalis (2).These results are supported by the fact that E. bieneusi and E. intestinalis are the most prevalent intestinal microsporida previously described in other countries including Africa , , , , , , , , , and E. cuniculi has only occasionally been found in the intestine of HIV-positive patients .
The PCR method identified 7 positive samples belonging to the species E. bieneusi and E. intestinalis (Table 2). The PCR technique is believed to be the most sensitive method for species identification, but it is expensive and not affordable in many clinical diagnostic laboratories. In the case of stool samples, the main problem is the appearance of false-negative results due to a low parasite-DNA concentration, and the presence of PCR inhibitors , , . Following from this, we should point out that we found a high presence of PCR inhibitors that had not been eliminated by the usual common methods , . This fact is supported by the number of control samples spiked with the cloned microsporidia DNA which were impossible to amplify. Their proportion was higher than those usually found in previous studies with fecal samples from other countries , . We can only speculate that the characteristics of the diets or treatments followed by these patients may contain PCR inhibitors. In addition to the above difficulties, it is also necessary to consider that human samples positive by the staining methods may not necessarily amplify with the specific primers used, due to other causes such as the possible presence of microsporidia other than the species studied, the overload of extruded spores in samples containing a low parasitic load, etc. The positive PCR results (3.6%) are much lower than those previously obtained, also by PCR, in Zimbabwean HIV-positive patients (51%) . However, in a study carried out in Mali, a country nearer Nigeria, E. bieneusi was found in 6.8% of HIV-positive patients and in 9% of HIV-negative individuals , and these data also correlates with the 9.3% of children recently identified in Nigeria as positive for E. bieneusi .
Few studies on microsporidia exist in Africa, but previous reports identified E. bieneusi as the most prevalent microsporidia infecting humans , , , , , , , . In the present report, E. bieneusi is described for the first time as the most prevalent microsporidia in HIV-positive patients with diarrhea in Nigeria. Additionally, E. intestinalis was also detected for the first time in this country although a previous report  described the presence of E. bieneusi/E. intestinalis but a without specific differentiation been performed. It is interesting to note the detection of E. cuniculi by IFAT in a fecal sample, although there are no previous references of this microsporidia in humans from the African continent. It is also noteworthy that in a study carried out in Nigeria in domestic rabbits, an E. cuniculi seropositivity of 16.5% was described, suggesting the zoonotic origin of this infection .
A high intraspecific variability in E. bieneusi has been described by several authors based on ITS of the rRNA gene. Most sequences found up to now for the ITS of E. bieneusi belong to Group I, including those found in our samples, and only a minority to divergent Groups II, III and IV . Recently, Breton et al.,  reported the prevalence in Cameroon and Gabon of several genotypes (A, B, D, K) included in Group I of the most common E. bieneusi haplotypes, and a divergent genotype CAF4 (Group II) whose significance is not yet clear . It has been suggested that some genotypes of E. bieneusi could possibly be host adapted genotypes and may be of importance to public health with reference to specific animal groups . However, a recent phylogenetic analysis showed that humans may become infected by E. bieneusi from different sources, including human-to-human, but there is also a certain degree of host-specificity . The sequences of the three Nigerian samples were similar to previously described genotypes found in E. bieneusi isolates from Africa, Europe, Asia and North and South America, and also from a number of different host species, including humans, cattle, pets and wild animals , , . According to their position in the network (Figure 1) they presumably represent the most ancient haplotypes of E. bieneusi. It is interesting to note that such central locations in the network apply to all the haplotypes found so far in Africa, including those recently reported by Ayinmode et al. . This result supports the idea that genotypes of E.bieneusi found in HIV-positive patients are closely related to those from several animals, thereby supporting the possibility of zoonotic transmission of the strains.
Finally, the prevalence of Cryptosporidium was established in 22.7% of the 193 fecal samples studied, with 47.7% and 9.5% in diarrheic and non-diarrheic samples respectively. These data are similar to those found for microsporidia using Trichrome stain (23.3% total, and 52.2% and 6.3% in diarrheic and non-diarrheic samples respectively), and are in accordance with previous reports indicating that Cryptosporidium infection is not the primary cause of diarrhea in HIV-positive patients in Africa , , . Moreover, microsporidia and Cryptosporidium were found coinfecting 30% of Cryptosporidium positive patients in this study, although this was not surprising as it is consistent with previous observations of both parasites in HIV/AIDS patients , , , , .
In conclusion, our results provide new data on microsporidia epidemiology on the African continent, where the control of HIV/AIDS is already due to limited access to HAART. Because of this, the presence of chronic intestinal microsporidiosis and cryptosporidiosis should be suspected in HIV/AIDS infected patients with persistent diarrhea, malabsorption and progressive weight loss. More importantly, there is further need to investigate other clinical and environmental samples in order to identify the major sources of these pathogens in human, in the environment.
Conceived and designed the experiments: OTO AFB NHG CdA. Performed the experiments: OTO FI. Analyzed the data: OTO FI SF AFB NHG CdA. Contributed reagents/materials/analysis tools: FI SF AFB WO AA NO NHG CdA. Wrote the paper: OTO FI SF AFB NHG CdA.
- 1. Serwadda D, Mugerwa RD, Sewankambo NK, Lwegaba A, Carswell JW, et al. (1985) Slim disease: a new disease in Uganda and its association with HTLV-III infection. Lancet 2: 849–852.D. SerwaddaRD MugerwaNK SewankamboA. LwegabaJW Carswell1985Slim disease: a new disease in Uganda and its association with HTLV-III infection.Lancet2849852
- 2. Didier ES, Stovall ME, Green LC, Brindley PJ, Sestak K, et al. (2004) Epidemiology of microsporidiosis: sources and modes of transmission. Vet Parasitol 126: 145–166.ES DidierME StovallLC GreenPJ BrindleyK. Sestak2004Epidemiology of microsporidiosis: sources and modes of transmission.Vet Parasitol126145166
- 3. Carlson JR, Li L, Helton CL, Munn RJ, Wasson K, et al. (2004) Disseminated microsporidiosis in a pancreas/kidney transplant recipient. Arch Pathol Lab Med 128: e41–43.JR CarlsonL. LiCL HeltonRJ MunnK. Wasson2004Disseminated microsporidiosis in a pancreas/kidney transplant recipient.Arch Pathol Lab Med128e4143
- 4. Mohindra AR, Lee MW, Visvesvara G, Moura H, Parasuraman R, et al. (2002) Disseminated microsporidiosis in a renal transplant recipient. Transpl Infect Dis 4: 102–107.AR MohindraMW LeeG. VisvesvaraH. MouraR. Parasuraman2002Disseminated microsporidiosis in a renal transplant recipient.Transpl Infect Dis4102107
- 5. Latib MA, Pascoe MD, Duffield MS, Kahn D (2001) Microsporidiosis in the graft of a renal transplant recipient. Transpl Int 14: 274–277.MA LatibMD PascoeMS DuffieldD. Kahn2001Microsporidiosis in the graft of a renal transplant recipient.Transpl Int14274277
- 6. Gomez Morales MA, Atzori C, Ludovisi A, Rossi P, Scaglia M, et al. (1995) Opportunistic and non-opportunistic parasites in HIV-positive and negative patients with diarrhoea in Tanzania. Trop Med Parasitol 46: 109–114.MA Gomez MoralesC. AtzoriA. LudovisiP. RossiM. Scaglia1995Opportunistic and non-opportunistic parasites in HIV-positive and negative patients with diarrhoea in Tanzania.Trop Med Parasitol46109114
- 7. Sadler F, Peake N, Borrow R, Rowl PL, Wilkins EG, et al. (2002) Genotyping of Enterocytozoon bieneusi in AIDS patients from the north west of England. J Infect 44: 39–42.F. SadlerN. PeakeR. BorrowPL RowlEG Wilkins2002Genotyping of Enterocytozoon bieneusi in AIDS patients from the north west of England.J Infect443942
- 8. Sing A, Tybus K, Heesemann J, Mathis A (2001) Molecular diagnosis of an Enterocytozoon bieneusi human genotype C infection in a moderately immunosuppressed human immunodeficiency virus seronegative liver-transplant recipient with severe chronic diarrhea. J Clin Microbiol 39: 2371–2372.A. SingK. TybusJ. HeesemannA. Mathis2001Molecular diagnosis of an Enterocytozoon bieneusi human genotype C infection in a moderately immunosuppressed human immunodeficiency virus seronegative liver-transplant recipient with severe chronic diarrhea.J Clin Microbiol3923712372
- 9. Metge S, Van Nhieu JT, Dahmane D, Grimbert P, Foulet F, et al. (2000) A case of Enterocytozoon bieneusi infection in an HIV-negative renal transplant recipient. Eur J Clin Microbiol Infect Dis 19: 221–223.S. MetgeJT Van NhieuD. DahmaneP. GrimbertF. Foulet2000A case of Enterocytozoon bieneusi infection in an HIV-negative renal transplant recipient.Eur J Clin Microbiol Infect Dis19221223
- 10. Galvan AL, Sanchez AM, Valentin MA, Henriques-Gil N, Izquierdo F, et al. (1301–1306) AL GalvanAM SanchezMA ValentinN. Henriques-GilF. Izquierdo1301–1306
- 11. Nkinin SW, Asonganyi T, Didier ES, Kaneshiro ES (2007) Microsporidian infection is prevalent in healthy people in Cameroon. J Clin Microbiol 45: 2841–2846.SW NkininT. AsonganyiES DidierES Kaneshiro2007Microsporidian infection is prevalent in healthy people in Cameroon.J Clin Microbiol4528412846
- 12. del Aguila C, Moura H, Fenoy S, Navajas R, Lopez-Velez R, et al. (2001) In vitro culture, ultrastructure, antigenic, and molecular characterization of Encephalitozoon cuniculi isolated from urine and sputum samples from a Spanish patient with AIDS. J Clin Microbiol 39: 1105–1108.C. del AguilaH. MouraS. FenoyR. NavajasR. Lopez-Velez2001In vitro culture, ultrastructure, antigenic, and molecular characterization of Encephalitozoon cuniculi isolated from urine and sputum samples from a Spanish patient with AIDS.J Clin Microbiol3911051108
- 13. Liguory O, David F, Sarfati C, Schuitema AR, Hartskeerl RA, et al. (1997) Diagnosis of infections caused by Enterocytozoon bieneusi and Encephalitozoon intestinalis using polymerase chain reaction in stool specimens. Aids 11: 723–726.O. LiguoryF. DavidC. SarfatiAR SchuitemaRA Hartskeerl1997Diagnosis of infections caused by Enterocytozoon bieneusi and Encephalitozoon intestinalis using polymerase chain reaction in stool specimens.Aids11723726
- 14. Kucerova-Pospisilova Z, Ditrich O (1998) The serological surveillance of several groups of patients using antigens of Encephalitozoon hellem and E. cuniculi antibodies to microsporidia in patients. Folia Parasitol (Praha) 45: 108–112.Z. Kucerova-PospisilovaO. Ditrich1998The serological surveillance of several groups of patients using antigens of Encephalitozoon hellem and E. cuniculi antibodies to microsporidia in patients.Folia Parasitol (Praha)45108112
- 15. van Gool T, Vetter JC, Weinmayr B, Van Dam A, Derouin F, et al. (1997) High seroprevalence of Encephalitozoon species in immunocompetent subjects. J Infect Dis 175: 1020–1024.T. van GoolJC VetterB. WeinmayrA. Van DamF. Derouin1997High seroprevalence of Encephalitozoon species in immunocompetent subjects.J Infect Dis17510201024
- 16. Sak B, Kvac M, Kucerova Z, Kvetonova D, Sakova K (2011) Latent microsporidial infection in immunocompetent individuals - a longitudinal study. PLoS Negl Trop Dis 5: e1162.B. SakM. KvacZ. KucerovaD. KvetonovaK. Sakova2011Latent microsporidial infection in immunocompetent individuals - a longitudinal study.PLoS Negl Trop Dis5e1162
- 17. Aldras AM, Orenstein JM, Kotler DP, Shadduck JA, Didier ES (1994) Detection of microsporidia by indirect immunofluorescence antibody test using polyclonal and monoclonal antibodies. J Clin Microbiol 32: 608–612.AM AldrasJM OrensteinDP KotlerJA ShadduckES Didier1994Detection of microsporidia by indirect immunofluorescence antibody test using polyclonal and monoclonal antibodies.J Clin Microbiol32608612
- 18. Leelayoova S, Subrungruang I, Suputtamongkol Y, Worapong J, Petmitr PC, et al. (2006) Identification of genotypes of Enterocytozoon bieneusi from stool samples from human immunodeficiency virus-infected patients in Thailand. J Clin Microbiol 44: 3001–3004.S. LeelayoovaI. SubrungruangY. SuputtamongkolJ. WorapongPC Petmitr2006Identification of genotypes of Enterocytozoon bieneusi from stool samples from human immunodeficiency virus-infected patients in Thailand.J Clin Microbiol4430013004
- 19. Notermans DW, Peek R, de Jong MD, Wentink-Bonnema EM, Boom R, et al. (2005) Detection and identification of Enterocytozoon bieneusi and Encephalitozoon species in stool and urine specimens by PCR and differential hybridization. J Clin Microbiol 43: 610–614.DW NotermansR. PeekMD de JongEM Wentink-BonnemaR. Boom2005Detection and identification of Enterocytozoon bieneusi and Encephalitozoon species in stool and urine specimens by PCR and differential hybridization.J Clin Microbiol43610614
- 20. Subrungruang I, Mungthin M, Chavalitshewinkoon-Petmitr P, Rangsin R, Naaglor T, et al. (2004) Evaluation of DNA extraction and PCR methods for detection of Enterocytozoon bienuesi in stool specimens. J Clin Microbiol 42: 3490–3494.I. SubrungruangM. MungthinP. Chavalitshewinkoon-PetmitrR. RangsinT. Naaglor2004Evaluation of DNA extraction and PCR methods for detection of Enterocytozoon bienuesi in stool specimens.J Clin Microbiol4234903494
- 21. Tumwine JK, Kekitiinwa A, Nabukeera N, Akiyoshi DE, Buckholt MA, et al. (2002) Enterocytozoon bieneusi among children with diarrhea attending Mulago Hospital in Uganda. Am J Trop Med Hyg 67: 299–303.JK TumwineA. KekitiinwaN. NabukeeraDE AkiyoshiMA Buckholt2002Enterocytozoon bieneusi among children with diarrhea attending Mulago Hospital in Uganda.Am J Trop Med Hyg67299303
- 22. Muller A, Bialek R, Kamper A, Fatkenheuer G, Salzberger B, et al. (2001) Detection of microsporidia in travelers with diarrhea. J Clin Microbiol 39: 1630–1632.A. MullerR. BialekA. KamperG. FatkenheuerB. Salzberger2001Detection of microsporidia in travelers with diarrhea.J Clin Microbiol3916301632
- 23. Franzen C, Muller A (1999) Molecular techniques for detection, species differentiation, and phylogenetic analysis of microsporidia. Clin Microbiol Rev 12: 243–285.C. FranzenA. Muller1999Molecular techniques for detection, species differentiation, and phylogenetic analysis of microsporidia.Clin Microbiol Rev12243285
- 24. Bern C, Kawai V, Vargas D, Rabke-Verani J, Williamson J, et al. (2005) The epidemiology of intestinal microsporidiosis in patients with HIV/AIDS in Lima, Peru. J Infect Dis 191: 1658–1664.C. BernV. KawaiD. VargasJ. Rabke-VeraniJ. Williamson2005The epidemiology of intestinal microsporidiosis in patients with HIV/AIDS in Lima, Peru.J Infect Dis19116581664
- 25. Mathis A, Breitenmoser AC, Deplazes P (1999) Detection of new Enterocytozoon genotypes in faecal samples of farm dogs and a cat. Parasite 6: 189–193.A. MathisAC BreitenmoserP. Deplazes1999Detection of new Enterocytozoon genotypes in faecal samples of farm dogs and a cat.Parasite6189193
- 26. Haro M, Izquierdo F, Henriques-Gil N, Andres I, Alonso F, et al. (2005) First detection and genotyping of human-associated microsporidia in pigeons from urban parks. Appl Environ Microbiol 71: 3153–3157.M. HaroF. IzquierdoN. Henriques-GilI. AndresF. Alonso2005First detection and genotyping of human-associated microsporidia in pigeons from urban parks.Appl Environ Microbiol7131533157
- 27. Haro M, Henriques-Gil N, Fenoy S, Izquierdo F, Alonso F, et al. (2006) Detection and genotyping of Enterocytozoon bieneusi in pigeons. J Eukaryot Microbiol 53: Suppl 1S58–60.M. HaroN. Henriques-GilS. FenoyF. IzquierdoF. Alonso2006Detection and genotyping of Enterocytozoon bieneusi in pigeons.J Eukaryot Microbiol53Suppl 1S5860
- 28. Bornay-Llinares FJ, da Silva AJ, Moura H, Schwartz DA, Visvesvara GS, et al. (1998) Immunologic, microscopic, and molecular evidence of Encephalitozoon intestinalis (Septata intestinalis) infection in mammals other than humans. J Infect Dis 178: 820–826.FJ Bornay-LlinaresAJ da SilvaH. MouraDA SchwartzGS Visvesvara1998Immunologic, microscopic, and molecular evidence of Encephalitozoon intestinalis (Septata intestinalis) infection in mammals other than humans.J Infect Dis178820826
- 29. Lobo ML, Xiao L, Cama V, Magalhaes N, Antunes F, et al. (2006) Identification of potentially human-pathogenic Enterocytozoon bieneusi genotypes in various birds. Appl Environ Microbiol 72: 7380–7382.ML LoboL. XiaoV. CamaN. MagalhaesF. Antunes2006Identification of potentially human-pathogenic Enterocytozoon bieneusi genotypes in various birds.Appl Environ Microbiol7273807382
- 30. Santin M, Trout JM, Fayer R (2005) Enterocytozoon bieneusi genotypes in dairy cattle in the eastern United States. Parasitol Res 97: 535–538.M. SantinJM TroutR. Fayer2005Enterocytozoon bieneusi genotypes in dairy cattle in the eastern United States.Parasitol Res97535538
- 31. Haro M, Del Aguila C, Fenoy S, Henriques-Gil N (2003) Intraspecies genotype variability of the microsporidian parasite Encephalitozoon hellem. J Clin Microbiol 41: 4166–4171.M. HaroC. Del AguilaS. FenoyN. Henriques-Gil2003Intraspecies genotype variability of the microsporidian parasite Encephalitozoon hellem.J Clin Microbiol4141664171
- 32. Henriques-Gil N, Haro M, Izquierdo F, Fenoy S, del Aguila C (2010) Phylogenetic approach to the variability of the microsporidian Enterocytozoon bieneusi and its implications for inter- and intrahost transmission. Appl Environ Microbiol 76: 3333–3342.N. Henriques-GilM. HaroF. IzquierdoS. FenoyC. del Aguila2010Phylogenetic approach to the variability of the microsporidian Enterocytozoon bieneusi and its implications for inter- and intrahost transmission.Appl Environ Microbiol7633333342
- 33. Ferreira FM, Bezerra L, Santos MB, Bernardes RM, Avelino I, et al. (2001) Intestinal microsporidiosis: a current infection in HIV-seropositive patients in Portugal. Microbes Infect 3: 1015–1019.FM FerreiraL. BezerraMB SantosRM BernardesI. Avelino2001Intestinal microsporidiosis: a current infection in HIV-seropositive patients in Portugal.Microbes Infect310151019
- 34. Samie A, Obi CL, Tzipori S, Weiss LM, Guerrant RL (2007) Microsporidiosis in South Africa: PCR detection in stool samples of HIV-positive and HIV-negative individuals and school children in Vhembe district, Limpopo Province. Trans R Soc Trop Med Hyg 101: 547–554.A. SamieCL ObiS. TziporiLM WeissRL Guerrant2007Microsporidiosis in South Africa: PCR detection in stool samples of HIV-positive and HIV-negative individuals and school children in Vhembe district, Limpopo Province.Trans R Soc Trop Med Hyg101547554
- 35. Sarfati C, Bourgeois A, Menotti J, Liegeois F, Moyou-Somo R, et al. (2006) Prevalence of intestinal parasites including microsporidia in human immunodeficiency virus-infected adults in Cameroon: a cross-sectional study. Am J Trop Med Hyg 74: 162–164.C. SarfatiA. BourgeoisJ. MenottiF. LiegeoisR. Moyou-Somo2006Prevalence of intestinal parasites including microsporidia in human immunodeficiency virus-infected adults in Cameroon: a cross-sectional study.Am J Trop Med Hyg74162164
- 36. Endeshaw T, Kebede A, Verweij JJ, Zewide A, Tsige K, et al. (2006) Intestinal microsporidiosis in diarrheal patients infected with human immunodeficiency virus-1 in Addis Ababa, Ethiopia. Jpn J Infect Dis 59: 306–310.T. EndeshawA. KebedeJJ VerweijA. ZewideK. Tsige2006Intestinal microsporidiosis in diarrheal patients infected with human immunodeficiency virus-1 in Addis Ababa, Ethiopia.Jpn J Infect Dis59306310
- 37. Sianongo S, McDonald V, Kelly P (2001) A method for diagnosis of microsporidiosis adapted for use in developing countries. Trans R Soc Trop Med Hyg 95: 605–607.S. SianongoV. McDonaldP. Kelly2001A method for diagnosis of microsporidiosis adapted for use in developing countries.Trans R Soc Trop Med Hyg95605607
- 38. Carville A, Mansfield K, Widmer G, Lackner A, Kotler D, et al. (1997) Development and application of genetic probes for detection of Enterocytozoon bieneusi in formalin-fixed stools and in intestinal biopsy specimens from infected patients. Clin Diagn Lab Immunol 4: 405–408.A. CarvilleK. MansfieldG. WidmerA. LacknerD. Kotler1997Development and application of genetic probes for detection of Enterocytozoon bieneusi in formalin-fixed stools and in intestinal biopsy specimens from infected patients.Clin Diagn Lab Immunol4405408
- 39. Kelly P, McPhail G, Ngwenya B, Luo N, Karew AH, et al. (1994) Septata intestinalis: a new microsporidian in Africa. Lancet 344: 271–272.P. KellyG. McPhailB. NgwenyaN. LuoAH Karew1994Septata intestinalis: a new microsporidian in Africa.Lancet344271272
- 40. van Gool T, Luderhoff E, Nathoo KJ, Kiire CF, Dankert J, et al. (1995) High prevalence of Enterocytozoon bieneusi infections among HIV-positive individuals with persistent diarrhoea in Harare, Zimbabwe. Trans R Soc Trop Med Hyg 89: 478–480.T. van GoolE. LuderhoffKJ NathooCF KiireJ. Dankert1995High prevalence of Enterocytozoon bieneusi infections among HIV-positive individuals with persistent diarrhoea in Harare, Zimbabwe.Trans R Soc Trop Med Hyg89478480
- 41. Ikeh EI, Obadofin MO, Brindeiro B, Baugherb C, Frost F, et al. (2007) Intestinal parasitism in Magama Gumau rural village and Jos township in north central Nigeria. Niger Postgrad Med J 14: 290–295.EI IkehMO ObadofinB. BrindeiroC. BaugherbF. Frost2007Intestinal parasitism in Magama Gumau rural village and Jos township in north central Nigeria.Niger Postgrad Med J14290295
- 42. Espern A, Morio F, Miegeville M, Illa H, Abdoulaye M, et al. (2007) Molecular study of microsporidiosis due to Enterocytozoon bieneusi and Encephalitozoon intestinalis among human immunodeficiency virus-infected patients from two geographical areas: Niamey, Niger, and Hanoi, Vietnam. J Clin Microbiol 45: 2999–3002.A. EspernF. MorioM. MiegevilleH. IllaM. Abdoulaye2007Molecular study of microsporidiosis due to Enterocytozoon bieneusi and Encephalitozoon intestinalis among human immunodeficiency virus-infected patients from two geographical areas: Niamey, Niger, and Hanoi, Vietnam.J Clin Microbiol4529993002
- 43. Breton J, Bart-Delabesse E, Biligui S, Carbone A, Seiller X, et al. (2007) New highly divergent rRNA sequence among biodiverse genotypes of Enterocytozoon bieneusi strains isolated from humans in Gabon and Cameroon. J Clin Microbiol 45: 2580–2589.J. BretonE. Bart-DelabesseS. BiliguiA. CarboneX. Seiller2007New highly divergent rRNA sequence among biodiverse genotypes of Enterocytozoon bieneusi strains isolated from humans in Gabon and Cameroon.J Clin Microbiol4525802589
- 44. Chabchoub N, Abdelmalek R, Mellouli F, Kanoun F, Thellier M, et al. (2009) Genetic identification of intestinal microsporidia species in immunocompromised patients in Tunisia. Am J Trop Med Hyg 80: 24–27.N. ChabchoubR. AbdelmalekF. MellouliF. KanounM. Thellier2009Genetic identification of intestinal microsporidia species in immunocompromised patients in Tunisia.Am J Trop Med Hyg802427
- 45. ten Hove RJ, Van Lieshout L, Beadsworth MB, Perez MA, Spee K, et al. (2009) Characterization of genotypes of Enterocytozoon bieneusi in immunosuppressed and immunocompetent patient groups. J Eukaryot Microbiol 56: 388–393.RJ ten HoveL. Van LieshoutMB BeadsworthMA PerezK. Spee2009Characterization of genotypes of Enterocytozoon bieneusi in immunosuppressed and immunocompetent patient groups.J Eukaryot Microbiol56388393
- 46. Ayinmode AB, Ojuromi OT, Xiao L Molecular Identification of Enterocytozoon bieneusi Isolates from Nigerian ChildrenJParasitolRes2011:129542 (9542) AB AyinmodeOT OjuromiChildrenJParasitolRes2011:129542 Xiao L Molecular Identification of Enterocytozoon bieneusi Isolates from Nigerian9542
- 47. Weber R, Bryan RT, Owen RL, Wilcox CM, Gorelkin L, et al. (1992) Improved light-microscopical detection of microsporidia spores in stool and duodenal aspirates. The Enteric Opportunistic Infections Working Group. N Engl J Med 326: 161–166.R. WeberRT BryanRL OwenCM WilcoxL. Gorelkin1992Improved light-microscopical detection of microsporidia spores in stool and duodenal aspirates. The Enteric Opportunistic Infections Working Group.N Engl J Med326161166
- 48. García LS (2007) Diagnostic Medical Parasitology. Washington, DC: American Society of Microbiology. LS García2007Diagnostic Medical Parasitology.Washington, DCAmerican Society of Microbiology
- 49. Del Aguila C, Navajas R, Gurbindo D, Ramos JT, Mellado MJ, et al. (1997) Microsporidiosis in HIV-positive children in Madrid (Spain). J Eukaryot Microbiol 44: 84S–85S.C. Del AguilaR. NavajasD. GurbindoJT RamosMJ Mellado1997Microsporidiosis in HIV-positive children in Madrid (Spain).J Eukaryot Microbiol4484S85S
- 50. del Aguila C, Izquierdo F, Navajas R, Pieniazek NJ, Miro G, et al. (1999) Enterocytozoon bieneusi in animals: rabbits and dogs as new hosts. J Eukaryot Microbiol 46: 8S–9S.C. del AguilaF. IzquierdoR. NavajasNJ PieniazekG. Miro1999Enterocytozoon bieneusi in animals: rabbits and dogs as new hosts.J Eukaryot Microbiol468S9S
- 51. Gainzarain JC, Canut A, Lozano M, Labora A, Carreras F, et al. (1998) Detection of Enterocytozoon bieneusi in two human immunodeficiency virus-negative patients with chronic diarrhea by polymerase chain reaction in duodenal biopsy specimens and review. Clin Infect Dis 27: 394–398.JC GainzarainA. CanutM. LozanoA. LaboraF. Carreras1998Detection of Enterocytozoon bieneusi in two human immunodeficiency virus-negative patients with chronic diarrhea by polymerase chain reaction in duodenal biopsy specimens and review.Clin Infect Dis27394398
- 52. Accoceberry I, Thellier M, Desportes-Livage I, Achbarou A, Biligui S, et al. (1999) Production of monoclonal antibodies directed against the microsporidium Enterocytozoon bieneusi. J Clin Microbiol 37: 4107–4112.I. AccoceberryM. ThellierI. Desportes-LivageA. AchbarouS. Biligui1999Production of monoclonal antibodies directed against the microsporidium Enterocytozoon bieneusi.J Clin Microbiol3741074112
- 53. da Silva AJ, Schwartz DA, Visvesvara GS, de Moura H, Slemenda SB, et al. (1996) Sensitive PCR diagnosis of Infections by Enterocytozoon bieneusi (microsporidia) using primers based on the region coding for small-subunit rRNA. J Clin Microbiol 34: 986–987.AJ da SilvaDA SchwartzGS VisvesvaraH. de MouraSB Slemenda1996Sensitive PCR diagnosis of Infections by Enterocytozoon bieneusi (microsporidia) using primers based on the region coding for small-subunit rRNA.J Clin Microbiol34986987
- 54. Da Silva AJ, Slemenda SB, Visvesvara GS, Schwartz DA, Wilcox CM, et al. (1997) Detection of Septata intestinalis (Microsporidia) Cali et al. 1993 Using Polymerase Chain Reaction Primers Targeting the Small Submit Subunit Ribosomal RNA Coding Region. Mol Diagn 2: 47–52.AJ Da SilvaSB SlemendaGS VisvesvaraDA SchwartzCM Wilcox1997Detection of Septata intestinalis (Microsporidia) Cali et al. 1993 Using Polymerase Chain Reaction Primers Targeting the Small Submit Subunit Ribosomal RNA Coding Region.Mol Diagn24752
- 55. Visvesvara GS, Leitch GJ, da Silva AJ, Croppo GP, Moura H, et al. (1994) Polyclonal and monoclonal antibody and PCR-amplified small-subunit rRNA identification of a microsporidian, Encephalitozoon hellem, isolated from an AIDS patient with disseminated infection. J Clin Microbiol 32: 2760–2768.GS VisvesvaraGJ LeitchAJ da SilvaGP CroppoH. Moura1994Polyclonal and monoclonal antibody and PCR-amplified small-subunit rRNA identification of a microsporidian, Encephalitozoon hellem, isolated from an AIDS patient with disseminated infection.J Clin Microbiol3227602768
- 56. del Aguila C, Lopez-Velez R, Fenoy S, Turrientes C, Cobo J, et al. (1997) Identification of Enterocytozoon bieneusi spores in respiratory samples from an AIDS patient with a 2-year history of intestinal microsporidiosis. J Clin Microbiol 35: 1862–1866.C. del AguilaR. Lopez-VelezS. FenoyC. TurrientesJ. Cobo1997Identification of Enterocytozoon bieneusi spores in respiratory samples from an AIDS patient with a 2-year history of intestinal microsporidiosis.J Clin Microbiol3518621866
- 57. Hall TA (1999) BioEdit: a user-friendly biological sequence añlignment editor and analysis program for Windows 95/98/NT. Nucl Acids Symp Ser 41: 95–98.TA Hall1999BioEdit: a user-friendly biological sequence añlignment editor and analysis program for Windows 95/98/NT.Nucl Acids Symp Ser419598
- 58. Thompson JD, Higgins DG, Gibson TJ (1994) CLAUSTAL W: improving the sensitivity of progressive multiple sequence alignments through sequence weighting, position- specific gap penalties and weight matrix choice. Nucl Acids Res 22: 4673–4680.JD ThompsonDG HigginsTJ Gibson1994CLAUSTAL W: improving the sensitivity of progressive multiple sequence alignments through sequence weighting, position- specific gap penalties and weight matrix choice.Nucl Acids Res2246734680
- 59. Henriques-Gil N, Haro M, Izquierdo F, Fenoy S, del Aguila C Phylogenetic approach to the variability of the microsporidian Enterocytozoon bieneusi, its implications for inter-, intrahost transmissionApplEnvironMicrobiol76:3333–3342N. Henriques-GilM. HaroF. IzquierdoS. Fenoybieneusi del Aguila C Phylogenetic approach to the variability of the microsporidian Enterocytozooninter- its implications fortransmissionApplEnvironMicrobiol76:3333–3342 intrahost
- 60. Rozas J, Sanchez-Del Barrio JC, Messeguer X, Rozas R (2003) DnaSP, DNA polymorphism analyses by the coalescent and other methods. Bioinformatics 19: 2496–2497.J. RozasJC Sanchez-Del BarrioX. MesseguerR. Rozas2003DnaSP, DNA polymorphism analyses by the coalescent and other methods.Bioinformatics1924962497
- 61. Bandelt HJ, Forster P, Rohl A (1999) Median-joining networks for inferring intraspecific phylogenies. Mol Biol Evol 16: 37–48.HJ BandeltP. ForsterA. Rohl1999Median-joining networks for inferring intraspecific phylogenies.Mol Biol Evol163748
- 62. Foster P (2001) Phylogenetic star contraction applied to Asian and Papua mtDNA evolution. Molecular Biology and Evolution 18: 1864–1881.P. Foster2001Phylogenetic star contraction applied to Asian and Papua mtDNA evolution.Molecular Biology and Evolution1818641881
- 63. Santin M, Fayer R (2009) Enterocytozoon bieneusi genotype nomenclature based on the internal transcribed spacer sequence: a consensus. J Eukaryot Microbiol 56: 34–38.M. SantinR. Fayer2009Enterocytozoon bieneusi genotype nomenclature based on the internal transcribed spacer sequence: a consensus.J Eukaryot Microbiol563438
- 64. Stark D, Barratt JL, van Hal S, Marriott D, Harkness J, et al. (2009) Clinical significance of enteric protozoa in the immunosuppressed human population. Clin Microbiol Rev 22: 634–650.D. StarkJL BarrattS. van HalD. MarriottJ. Harkness2009Clinical significance of enteric protozoa in the immunosuppressed human population.Clin Microbiol Rev22634650
- 65. Tumwine JK, Kekitiinwa A, Bakeera-Kitaka S, Ndeezi G, Downing R, et al. (2005) Cryptosporidiosis and microsporidiosis in ugandan children with persistent diarrhea with and without concurrent infection with the human immunodeficiency virus. Am J Trop Med Hyg 73: 921–925.JK TumwineA. KekitiinwaS. Bakeera-KitakaG. NdeeziR. Downing2005Cryptosporidiosis and microsporidiosis in ugandan children with persistent diarrhea with and without concurrent infection with the human immunodeficiency virus.Am J Trop Med Hyg73921925
- 66. Didier ES (2005) Microsporidiosis: an emerging and opportunistic infection in humans and animals. Acta Trop 94: 61–76.ES Didier2005Microsporidiosis: an emerging and opportunistic infection in humans and animals.Acta Trop946176
- 67. Gumbo T, Sarbah S, Gangaidzo IT, Ortega Y, Sterling CR, et al. (1999) Intestinal parasites in patients with diarrhea and human immunodeficiency virus infection in Zimbabwe. Aids 13: 819–821.T. GumboS. SarbahIT GangaidzoY. OrtegaCR Sterling1999Intestinal parasites in patients with diarrhea and human immunodeficiency virus infection in Zimbabwe.Aids13819821
- 68. Lebbad M, Norrgren H, Naucler A, Dias F, Andersson S, et al. (2001) Intestinal parasites in HIV-2 associated AIDS cases with chronic diarrhoea in Guinea-Bissau. Acta Trop 80: 45–49.M. LebbadH. NorrgrenA. NauclerF. DiasS. Andersson2001Intestinal parasites in HIV-2 associated AIDS cases with chronic diarrhoea in Guinea-Bissau.Acta Trop804549
- 69. Boom R, Sol CJ, Salimans MM, Jansen CL, Wertheim-van Dillen PM, et al. (1990) Rapid and simple method for purification of nucleic acids. J Clin Microbiol 28: 495–503.R. BoomCJ SolMM SalimansCL JansenPM Wertheim-van Dillen1990Rapid and simple method for purification of nucleic acids.J Clin Microbiol28495503
- 70. Fedorko DP, Nelson NA, Cartwright CP (1995) Identification of microsporidia in stool specimens by using PCR and restriction endonucleases. J Clin Microbiol 33: 1739–1741.DP FedorkoNA NelsonCP Cartwright1995Identification of microsporidia in stool specimens by using PCR and restriction endonucleases.J Clin Microbiol3317391741
- 71. Lores B, Lopez-Miragaya I, Arias C, Fenoy S, Torres J, et al. (2002) Intestinal microsporidiosis due to Enterocytozoon bieneusi in elderly human immunodeficiency virus-negative patients from Vigo, Spain. Clin Infect Dis 34: 918–921.B. LoresI. Lopez-MiragayaC. AriasS. FenoyJ. Torres2002Intestinal microsporidiosis due to Enterocytozoon bieneusi in elderly human immunodeficiency virus-negative patients from Vigo, Spain.Clin Infect Dis34918921
- 72. Maiga I, Doumbo O, Dembele M, Traore H, Desportes-Livage I, et al. (1997) [Human intestinal microsporidiosis in Bamako (Mali): the presence of Enterocytozoon bieneusi in HIV seropositive patients]. Sante 7: 257–262.I. MaigaO. DoumboM. DembeleH. TraoreI. Desportes-Livage1997[Human intestinal microsporidiosis in Bamako (Mali): the presence of Enterocytozoon bieneusi in HIV seropositive patients].Sante7257262
- 73. Bretagne S, Foulet F, Alkassoum W, Fleury-Feith J, Develoux M (1993) [Prevalence of Enterocytozoon bieneusi spores in the stool of AIDS patients and African children not infected by HIV]. Bull Soc Pathol Exot 86: 351–357.S. BretagneF. FouletW. AlkassoumJ. Fleury-FeithM. Develoux1993[Prevalence of Enterocytozoon bieneusi spores in the stool of AIDS patients and African children not infected by HIV].Bull Soc Pathol Exot86351357
- 74. Alfa Cisse O, Ouattara A, Thellier M, Accoceberry I, Biligui S, et al. (2002) Evaluation of an immunofluorescent-antibody test using monoclonal antibodies directed against Enterocytozoon bieneusi and Encephalitozoon intestinalis for diagnosis of intestinal microsporidiosis in Bamako (Mali). J Clin Microbiol 40: 1715–1718.O. Alfa CisseA. OuattaraM. ThellierI. AccoceberryS. Biligui2002Evaluation of an immunofluorescent-antibody test using monoclonal antibodies directed against Enterocytozoon bieneusi and Encephalitozoon intestinalis for diagnosis of intestinal microsporidiosis in Bamako (Mali).J Clin Microbiol4017151718
- 75. Mor SM, Tumwine JK, Naumova EN, Ndeezi G, Tzipori S (2009) Microsporidiosis and malnutrition in children with persistent diarrhea, Uganda. Emerg Infect Dis 15: 49–52.SM MorJK TumwineEN NaumovaG. NdeeziS. Tzipori2009Microsporidiosis and malnutrition in children with persistent diarrhea, Uganda.Emerg Infect Dis154952
- 76. Okewole EA (2008) Seroprevalence of antibodies to Encephalitozoon cuniculi in domestic rabbits in Nigeria. Onderstepoort J Vet Res 75: 33–38.EA Okewole2008Seroprevalence of antibodies to Encephalitozoon cuniculi in domestic rabbits in Nigeria.Onderstepoort J Vet Res753338
- 77. Xiao L, Sulaiman , IM , Cama , V , Gilman , RH (2004) Molecular Epidemiology of human microsporidiosis caused by Enterocytozoon bieneusi. Southeast Asian J Trop Med Public Health 35: 40–44.L. XiaoSulaimanIMCamaVGilmanRH2004Molecular Epidemiology of human microsporidiosis caused by Enterocytozoon bieneusi.Southeast Asian J Trop Med Public Health354044
- 78. Sulaiman IM, Bern C, Gilman R, Cama V, Kawai V, et al. (2003) A molecular biologic study of Enterocytozoon bieneusi in HIV-infected patients in Lima, Peru. J Eukaryot Microbiol 50 Suppl. pp. 591–596.IM SulaimanC. BernR. GilmanV. CamaV. Kawai2003A molecular biologic study of Enterocytozoon bieneusi in HIV-infected patients in Lima, Peru.J Eukaryot Microbiol 50 Suppl591596
- 79. Cegielski JP, Ortega YR, McKee S, Madden JF, Gaido L, et al. (1999) Cryptosporidium, enterocytozoon, and cyclospora infections in pediatric and adult patients with diarrhea in Tanzania. Clin Infect Dis 28: 314–321.JP CegielskiYR OrtegaS. McKeeJF MaddenL. Gaido1999Cryptosporidium, enterocytozoon, and cyclospora infections in pediatric and adult patients with diarrhea in Tanzania.Clin Infect Dis28314321
- 80. Garcia LS, Shimizu RY, Bruckner DA (1994) Detection of microsporidial spores in fecal specimens from patients diagnosed with cryptosporidiosis. J Clin Microbiol 32: 1739–1741.LS GarciaRY ShimizuDA Bruckner1994Detection of microsporidial spores in fecal specimens from patients diagnosed with cryptosporidiosis.J Clin Microbiol3217391741
- 81. Weber R, Sauer B, Luthy R, Nadal D (1993) Intestinal coinfection with Enterocytozoon bieneusi and Cryptosporidium in a human immunodeficiency virus-infected child with chronic diarrhea. Clin Infect Dis 17: 480–483.R. WeberB. SauerR. LuthyD. Nadal1993Intestinal coinfection with Enterocytozoon bieneusi and Cryptosporidium in a human immunodeficiency virus-infected child with chronic diarrhea.Clin Infect Dis17480483