Opportunistic intestinal parasites in immunocompromised patients from a tertiary hospital in Monterrey, Mexico.

Opportunistic parasites are still important agents causing morbidity and mortality in immunocompromised patients, particularly those living with HIV/AIDS. Few studies in Mexico have attempted to determine the prevalence of opportunistic intestinal parasites causing diarrhea in immunocompromised patients. A study was conducted to determine the intestinal parasites in HIV-positive and HIV-negative immunocompromised patients with diarrhea admitted to a tertiary care hospital in Monterrey, Mexico, from 2014 to 2015. Stool samples were examined for trophozoites, cysts, and eggs using the EGRoPe sedimentation-concentration technique and special techniques (modified Ziehl-Neelsen stain, modified trichrome stain). A total of 56 patients were included. The overall prevalence of intestinal parasitism was 64% (36/56); 22/36 patients were HIV-positive. Prevalence of opportunistic parasites was 69% in HIV-infected patients compared to 44% in HIV-negative patients (P = 0.06). Microsporidia were the most frequently identified parasites (24/36, 67%), followed by Cryptosporidium sp. (6/36, 17%), Sarcocystis sp. (4/36, 11%), Cystoisospora belli (3/36, 8%), and Cyclospora cayetanensis (1/36, 3%). Overall prevalence rates of microsporidiosis and cryptosporidiosis were 43% and 11%, respectively. Among HIV-infected patients, prevalence rates of microsporidiosis and cryptosporidiosis were 48% and 14%, respectively. We also report the first cases of intestinal sarcocystosis in Mexico, all in HIV-infected patients. In conclusion, microsporidia and coccidia are major parasitic agents causing diarrhea in immunocompromised patients, particularly HIV-infected patients.

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.

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

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.

Cryptosporidiosis occurrence in HIV+ patients attended in a hospital, Brazil.

Cryptosporidiosis occurrence was determined in HIV+ patient assisted in the Clinic of Infect-parasitic Diseases in a hospital of Nova Iguaçu, Rio de Janeiro, Brazil, in the period from July/1998 to March/1999. In order to research, seventy-five patient, carriers of diarrhea or not, were appraised. The samples of feces were collected and placed in saline solution with formaldehyde (5%). The Modified Ritchie technique was used for the oocysts research, and the smears were stained with Safranine O methylene blue. The results verified 9.33% of positive samples, with higher frequency of cases in patients of the masculine sex from 20 to 50 years old, however without significant difference.

[Diagnosis of coccidia and microspores in specimens of diarrheal feces from Cuban HIV seropositive patients: first report of microspores in Cuba]. / Diagnóstico de coccidias y microsporas en muestras de heces diarreicas de pacientes cubanos seropositivos al VIH: primer reporte de microsporas en Cuba.

The numerous specimens of liquid diarrheas received at the clinical parasitology laboratory of "Pedro Kouri" Institute of Tropical Medicine were studied searching for the presence of pathogenic agents of Coccidium subclass and of the microspore family. The conditions to diagnose microsporidia were created on July 13, 2000, and the method was incorporated to the battery of diagnostic techniques used in the laboratory. A cut-off was made after 4 months to analyze the positivity found. A frequency distribution study was conducted and it was observed that of the 170 studied samples, 51 proved to be positive for some protozoon, which represents 30% of positivity. The most frequently found species were Cryptosporidium parvum and microsporidium. Association of various species in a same sample was found in 13.1% of the positive samples. The most frequent associations were: Cryptosporidium parvum and Cyclospora cayetanensis, Cryptosporidium parvum and microsporidium, and Cyclospora cayetanensis and microsporidium, each with 2 patients. Association of 3 different species in a same sample was also found. All the individuals in the series were HIV seropositive and the most affected had less than 200 TCD4+ lymphocytes/mL. This was the first report of microsporidia in feces in Cuba. It was possible after applying and interpreting the technique previously described to identify this phylum. As there is a trend towards the increase of cases with HIV/AIDS infection, it is necessary that the laboratories have the indispendable reagents to perform the modified Didier's trichromic staining and that the professionals and technicians of the parasitology laboratories of the country be trained in the procedures for identifying and recognizing these opportunistic protozoa.

Cryptosporidium species and genotypes in HIV-positive patients in Lima, Peru.

Cryptosporidium parasites from a cross-sectional study conducted in two national hospitals in Lima, Peru were genetically characterized to determine the diversity of Cryptosporidium spp. in HIV-positive people. A total of 2,672 patients participated in this study and provided 13,937 specimens. Cryptosporidium oocysts were detected by microscopy in 354 (13.3%) of the patients. Analysis of 951 Cryptosporidium-positive specimens from 300 patients using a small subunit rRNA-based PCR-RFLP tool identified 6 genotypes; Cryptosporidium hominis was the species most frequently detected (67.5%), followed by C. meleagridis (12.6%) and C. parvum (11.3%). Cryptosporidium canis (4.0%), C. felis (3.3%), and Cryptosporidium pig genotype (0.5%) were also found. These findings indicate that C. hominis is the predominant species in Peruvian HIV-positive persons, and that zoonotic Cryptosporidium spp. account for about 30% of cryptosporidiosis in these patients.

The relevance of laboratory diagnosis of human cryptosporidiosis and other coccidia.

Human infection by Cryptosporidium spp and other coccidia are due to opportunist non-host specific microorganisms. In HIV seropositive patients, the gastrointestinal symptoms accompanying such infections may be serious and prolonged and may include nausea, low-grade fever, abdominal cramps, anorexia and watery diarrhoea. We studied 188 stool samples from 111 patients (84 men and 27 women) with diarrhoea. A modified Ziehl-Nielsen technique for the detection of Cryptosporidium spp and Isospora belli was employed. The mean age of the patients was 31 years. Cryptosporidium spp was seen in 18% (n = 20) of the patients, 90% (n = 18) of whom were HIV seropositive. Isospora belli was recorded only from HIV seropositive patients (5.4% of all the patients studied and 6.5% of those who were HIV seropositive). These data confirm the good results obtained with this technique for the identification of Cryptosporidium spp and other coccidia and also reaffirm the clinical importance of correctly diagnosing the cause of diarrhoea, particularly in HIV seropositive patients.

[Comparison of intestinal parasite infestation indexes among HIV positive and negative populations]. / Comparación de índices de infestaciones por enteroparásitos entre poblaciones HIV positivas y negativas.

This study was carried out on two groups of patients treated at the José de San Martín Clinical Hospital. One group was composed of 82 HIV-seropositive patients with no signs of diarrhea, and another one of 300 patients, not suspected of HIV infection, was considered as "control population". Stool samples were collected from each patient and examined for intestinal parasites. These determinations were carried out at the Department of Clinical Biochemistry, in the same Hospital. Three specimens were obtained from each patient and processed in an identical form. Concentration methods, a direct smear technique after centrifugation and the trichrome staining procedure were performed on each sample. The HIV-positive group was found to harbor a higher proportion of intestinal parasites. Outstanding differences between both groups in percentages of parasitoses were observed. Species found in a significantly higher percentage among HIV seropositive patients were: Entamoeba histolytica (26.5%), Iodamoeba bütschlii (16.9%), Dientamoeba fragilis (25.3%), Blastocystis hominis (51.8%), Cryptosporidium sp. (7.2%), and probably Isospora belli (1.2%). No significant differences in percentages of Giardia lamblia, Entamoeba coli, Chilomastix mesnili and helminths were observed between both groups. Within the HIV-positive group, a higher proportion of infestation due to E. histolytica and I. bütschlii was observed in homosexuals-bisexuals than in intravenous drug addicts or heterosexuals.

Hallazgos de parásitos en pacientes HIV seropositivos / Parasites found in HIV seropositive patients

Durante 41 meses se investigó la presencia de parásitos en una población HIV seropositiva con las características clínicas de los estadíos 3 y 4 según la clasificación de la OMS. Se analizaron 212 muestras fecales de 135 pacientes y 53,33 por ciento de ellos presentaban enteroparásitos. A las heces recolectadas en formol 10 por ciento se les realizó un examen parasitológico directo, una concentración por Ritchie y la tinción de 2 frotis con Safranina 1 por ciento y 2 con Ziehl-Neelsen modificada para identificar Cryptosporidium sp. Las frecuencias detectadas fueron: Cryptosporidium sp. 11,11 por ciento; I. belli 2,96 por ciento; G. lamblia 11,85 por ciento; B. hominis 26,66 por ciento; A. lumbricoides 2,96 por ciento; E. vermicularis 1,48 por ciento; H. nana 0,74 por ciento; E. coli 13,33 por ciento por ciento; E. nana 5,93 por ciento; Ch. mesnilii 2,22 por ciento e I. butschlii 0,74 por ciento; resultando 46 pacientes monoparasitados, 19 b, 5 tri y 2 tetraparasitados. Se procesaron 17 lavados broncoalveolares (LBA) y 194 esputos, recolectados en formal 10 por ciento y centrifugados hasta agotamiento. Con el sedimento se realizaron 10 frotis y se colorearon con azul de toluidina O. La tinción de Groccot Gomori se usó para confirmar los casos dudosos. En el 47 por ciento de los LBA y en 22,68 por ciento de los esputos (E) se diagnosticó P. carinii. Esto representa el 34,68 por ciento. El porcentaje de positividad fue: 30,88 por ciento para los pacientes que remitieron un único esputo; 36,84 por ciento más de uno y 27,276 por ciento para LBA. Por último 7 pacientes enviaron LBA y E, siendo 2 positivos y 2 negativos en ambos materiales, mientras que en 3 pacientes se diagnosticó P. carinii sólo en LBA

Hallazgos de parásitos en pacientes HIV seropositivos / Parasites found in HIV seropositive patients

Durante 41 meses se investigó la presencia de parásitos en una población HIV seropositiva con las características clínicas de los estadíos 3 y 4 según la clasificación de la OMS. Se analizaron 212 muestras fecales de 135 pacientes y 53,33 por ciento de ellos presentaban enteroparásitos. A las heces recolectadas en formol 10 por ciento se les realizó un examen parasitológico directo, una concentración por Ritchie y la tinción de 2 frotis con Safranina 1 por ciento y 2 con Ziehl-Neelsen modificada para identificar Cryptosporidium sp. Las frecuencias detectadas fueron: Cryptosporidium sp. 11,11 por ciento; I. belli 2,96 por ciento; G. lamblia 11,85 por ciento; B. hominis 26,66 por ciento; A. lumbricoides 2,96 por ciento; E. vermicularis 1,48 por ciento; H. nana 0,74 por ciento; E. coli 13,33 por ciento por ciento; E. nana 5,93 por ciento; Ch. mesnilii 2,22 por ciento e I. butschlii 0,74 por ciento; resultando 46 pacientes monoparasitados, 19 b, 5 tri y 2 tetraparasitados. Se procesaron 17 lavados broncoalveolares (LBA) y 194 esputos, recolectados en formal 10 por ciento y centrifugados hasta agotamiento. Con el sedimento se realizaron 10 frotis y se colorearon con azul de toluidina O. La tinción de Groccot Gomori se usó para confirmar los casos dudosos. En el 47 por ciento de los LBA y en 22,68 por ciento de los esputos (E) se diagnosticó P. carinii. Esto representa el 34,68 por ciento. El porcentaje de positividad fue: 30,88 por ciento para los pacientes que remitieron un único esputo; 36,84 por ciento más de uno y 27,276 por ciento para LBA. Por último 7 pacientes enviaron LBA y E, siendo 2 positivos y 2 negativos en ambos materiales, mientras que en 3 pacientes se diagnosticó P. carinii sólo en LBA (AU)

[Parasites found in HIV-seropositive patients]. / Hallazgos de parásitos en pacientes HIV seropositivos.

For 41 months, the presence of parasites was investigated in a seropositive HIV population with the clinical characteristics of stages 3 and 4 according to the OMS classification; of 212 fecal samples belonging to 135 patients which were analyzed, 53.33% presented enteroparasites. A direct parasitological exam and a Ritchie concentration were performed on the feces collected in formol 10%. Two smears were stained with Safranine 1% and two with modified Ziehl-Nielsen to identify Cryptosporidium sp. The detected frequencies were: Cryptosporidium The detected frequencies were: Cryptosporidium sp. 11.11%; I. belli 2.96%; G. lamblia 11.85%; B. hominis 26.66%; A. lumbricoides 2.96%; E. vermicularis 1.48%; H. nana 0.74%; E. coli 13.33%; E. nana 5.93%; Ch. mesnilii 2.22% and I. butschlii 0.74%. There were 46 monoparasitized patients, 19 biparasitized, 5 triparasitized and 2 tetraparasitized. Furthermore, 17 bronchoalveolar lavages (BAL) and 194 sputa were processed, collected in formol 10% and centrifuged to exhaustion; 10 smears were prepared with sediment and were stained with toluidine blue. Groccot (Gomori) coloration was used to confirm doubtful cases. In 47% of the BAL and in 22,68% of the sputa P. carinii was diagnosed. This represents 34.68%. The percentage of positive cases was: 30.88% for those patients who sent a single sputum, 36.84% for those who sent more than one and 27.27% for BAL. Finally, in 7 patients who sent BAL and sputa, there were 2 positive and 2 negative cases in both materials, while P. carinii was diagnosed in 3 patients only in their BAL.