Un parásito emergente de patogenicidad controvertida, Blastocystis sp.: ¿Hay que tratar a todos los pacientes? / Blastocystis sp., an emerging parasite with controversial pathogenicity. Should all human cases be treated?

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Paludismo importado en adultos. Perfil clínico, epidemiológico y analítico / Imported malaria in adults. Clinical, epidemiological and analytical features

Introducción. Las características epidemiológicas y clínicas de los casos de paludismo importado en España se han descrito hasta ahora en pequeñas series hospitalarias. En casi todas ellas el diagnóstico se ha realizado a partir de episodios sintomáticos. El objetivo del presente estudio es conocer las características epidemiológicas, clínicas y analíticas del paludismo importado en una Unidad de Referencia para Enfermedades Tropicales. Pacientes y métodos. . Se realiza un estudio observacional de corte transversal retrospectivo. La serie está constituida por los enfermos diagnosticados de malaria en el Hospital Carlos III entre el 1 de enero de 2002 y el 31 de diciembre de 2007. Resultados. Se identificaron 484 episodios de paludismo, incluyéndose en el análisis 398 casos. Casi la mitad de los pacientes eran nativos de zonas endémicas, mientras que la otra mitad eran viajeros o nativos-viajeros. En la mayoría de los viajes a zonas endémicas (88-98% según los grupos) no se había realizado quimioprofilaxis antipalúdica de forma correcta. El 30,4% de los pacientes se encontraban asintomáticos en el momento del diagnóstico. El 28,1% de los asintomáticos tenían anemia, el 19,8% trombopenia, el 14% leucopenia, el 5% hipocolesterolemia, el 5% insuficiencia renal y el 4,1% hipoglucemia. El 97,5% de los individuos asintomáticos tenían parasitemia baja, frente al 80,5% de los sintomáticos (p<0,001). Discusión. La mayoría de los pacientes que viajan a zonas endémicas en los que se diagnostica paludismo no realizan una quimioprofilaxis correcta. Se debe descartar la existencia de malaria en individuos procedentes de regiones tropicales con síntomas compatibles, así como también en algunos individuos asintomáticos con alteraciones analíticas(AU)

Introduction. Up to now, the epidemiological and clinical features of imported malaria in Spain have been described in small series from general hospitals. Almost all diagnosis had been made based on symptomatic patients. The aim of this study has been to determine the epidemiological, clinical and laboratorial characteristics of imported malaria in a Reference Unit for Tropical Diseases. Patients and methods. We performed a cross-sectional, observational and retrospective study. The series consisted of patients diagnosed of malaria who had been attended at the Hospital Carlos III from January 1, 2002 to December 31, 2007. Results. We identified 484 episodes of malaria, of which 398 cases were included in the analysis. Almost 50% of the patients were natives of endemic areas, while the rest were native-travelers or travelers. Most cases (88-98% according to the group) had not taken malaria chemoprophylaxis correctly when indicated. At the time of diagnosis, 30.4% of patients were asymptomatic and 28.1% of asymptomatic patients had anemia, 19.8% thrombocytopenia, 14% leukopenia, 5% hypocholesterolemia, 5% renal failure and 4.1% hypoglycemia. Low parasitemia was present in 97.5% of asymptomatic individuals compared to 80.5% of the symptomatic patients (P<0.001). Discussion. Absence of chemoprophylaxis (or poor compliance) is the main reason for malaria in individuals traveling to endemic areas. Malaria must be ruled out in individuals coming from tropical countries with compatible symptoms, and it also should be suspected in certain groups of asymptomatic individuals with abnormal laboratorial parameters(AU)

Paludismo importado e infección por VIH en Madrid. Perfil clínico y epidemiológico / Imported malaria and HIV infection in Madrid. Clinical and epidemiological features

Introducción. En España disponemos de pocos datos referentes a pacientes con paludismo coinfectados por el virus de inmunodeficiencia humana (VIH). Este estudio pretende precisar las características epidemiológicas y clínicas del paludismo importado en pacientes coinfectados por el VIH en nuestro medio. Pacientes y métodos. Serie de casos retrospectiva, a partir de las historias clínicas. La población de estudio está constituida por los enfermos diagnosticados de malaria en nuestro centro entre el 1 de enero de 2002 y el 31 de diciembre de 2007. Resultados. Se identificaron 484 episodios de paludismo, incluyéndose en el estudio 398 casos. La coinfección con el VIH se describió en 32 casos. Todos ellos ocurrieron en individuos en los que presumiblemente existía algún grado de semiinmunidad. En el grupo de los coinfectados se encontraban asintomáticos 13 casos (40,6%), mientras que esta circunstancia se produjo en 99 casos de los pacientes no coinfectados (37,2%) (p=0,707). Destacó la mayor presencia de anemia en los pacientes coinfectados (62,5% en pacientes VIH frente a 32,3% en los no coinfectados [p=0,001]). Conclusiones. Las formas de presentación clínica del paludismo no parecen variar en función de la coexistencia con infección por el VIH. Si bien la población estudiada no reproduce todos los escenarios posibles de infección por VIH, señala la realidad de los pacientes que llegan a la Comunidad Autónoma de Madrid(AU)

Introduction. Few data are available in Spain data on human immunodeficiency virus (HIV) patients coinfected with malaria. This study has aimed to determine the epidemiological and clinical characteristics of imported malaria in patients coinfected with HIV. Patients and methods. A case-series retrospective study was performed using the patient's medical records. The study population consisted on patients diagnosed with malaria attended in our center from january 1, 2002 to december 31, 2007. Results. A total of 484 episodes of malaria, 398 of which were included in this study, were identified. Co-infection with HIV was described in 32 cases. All of them occurred in individuals presumably with some degree of semi-immunity. In the coinfected group, there were 13 cases (40.6%) asymptomatic, whereas this event occurred in 99 cases of patients not coinfected (37.2%) (P=0.707). The greater presence of anemia in co-infected patients (62.5% vs 32.3% in non-coinfected [P=0.001]) stands out. Conclusions. In present study, the clinical presentation forms were similar, regardless of the presence or absence of HIV infection. Although the study population does not reflect all possible scenarios of malaria and HIV coinfection, our results indicate the reality of patients attended in the Autonomous Community of Madrid(AU)

[Imported malaria and HIV infection in Madrid. Clinical and epidemiological features]. / Paludismo importado e infección por VIH en Madrid. Perfil clínico y epidemiológico.

INTRODUCTION: Few data are available in Spain data on human immunodeficiency virus (HIV) patients coinfected with malaria. This study has aimed to determine the epidemiological and clinical characteristics of imported malaria in patients coinfected with HIV. PATIENTS AND METHODS: A case-series retrospective study was performed using the patient's medical records. The study population consisted on patients diagnosed with malaria attended in our center from january 1, 2002 to december 31, 2007. RESULTS: A total of 484 episodes of malaria, 398 of which were included in this study, were identified. Co-infection with HIV was described in 32 cases. All of them occurred in individuals presumably with some degree of semi-immunity. In the coinfected group, there were 13 cases (40.6%) asymptomatic, whereas this event occurred in 99 cases of patients not coinfected (37.2%) (P=0.707). The greater presence of anemia in co-infected patients (62.5% vs 32.3% in non-coinfected [P=0.001]) stands out. CONCLUSIONS: In present study, the clinical presentation forms were similar, regardless of the presence or absence of HIV infection. Although the study population does not reflect all possible scenarios of malaria and HIV coinfection, our results indicate the reality of patients attended in the Autonomous Community of Madrid.

[Imported malaria in adults. Clinical, epidemiological and analytical features]. / Paludismo importado en adultos. Perfil clínico, epidemiológico y analítico.

INTRODUCTION: Up to now, the epidemiological and clinical features of imported malaria in Spain have been described in small series from general hospitals. Almost all diagnosis had been made based on symptomatic patients. The aim of this study has been to determine the epidemiological, clinical and laboratorial characteristics of imported malaria in a Reference Unit for Tropical Diseases. PATIENTS AND METHODS: We performed a cross-sectional, observational and retrospective study. The series consisted of patients diagnosed of malaria who had been attended at the Hospital Carlos III from January 1, 2002 to December 31, 2007. RESULTS: We identified 484 episodes of malaria, of which 398 cases were included in the analysis. Almost 50% of the patients were natives of endemic areas, while the rest were native-travelers or travelers. Most cases (88-98% according to the group) had not taken malaria chemoprophylaxis correctly when indicated. At the time of diagnosis, 30.4% of patients were asymptomatic and 28.1% of asymptomatic patients had anemia, 19.8% thrombocytopenia, 14% leukopenia, 5% hypocholesterolemia, 5% renal failure and 4.1% hypoglycemia. Low parasitemia was present in 97.5% of asymptomatic individuals compared to 80.5% of the symptomatic patients (P<0.001). DISCUSSION: Absence of chemoprophylaxis (or poor compliance) is the main reason for malaria in individuals traveling to endemic areas. Malaria must be ruled out in individuals coming from tropical countries with compatible symptoms, and it also should be suspected in certain groups of asymptomatic individuals with abnormal laboratorial parameters.

Application of real-time PCR for the differentiation of Entamoeba histolytica and E. dispar in cyst-positive faecal samples from 130 immigrants living in Spain.

In an effort to improve the diagnosis of intestinal amoebiasis, a real-time PCR has been used for the detection and differentiation of Entamoeba histolytica and E. dispar infections in African or South American immigrants who live in Spain. Faecal samples from all of the 130 subjects had apparently been found to contain E. histolytica/E. dispar cysts by microscopical examination. Using the real-time PCR, E. histolytica DNA was detected in faecal samples from only 10 (7.7%) of the immigrants, with E. dispar DNA detected in the samples from another 117 (90.0%) of the subjects. The use of such PCR in the routine investigation of patients found positive for E. histolytica/E. dispar cysts (by microscopy) is recommended, especially in non-endemic areas.

Abnormal depolarizing patterns in three patients with filarial infection.

Several authors have described a particular potential of automated depolarization analysis in detecting malaria infection as part of the routine full blood count (FBC) performed by the Cell-Dyn 4000 analyzer. In these cases, abnormal depolarizing patterns are due to the presence of leukocyte-associated malaria hemozoin, a pigment which depolarizes the laser light. In this report we describe samples from three individual patients who did not have malaria infection but showed abnormal depolarizing events. Further investigation determined that these samples were from patients infected by the nematode Mansonella perstans. The observed depolarizing pattern consisted of a normal depolarizing eosinophil population and in addition an abnormal depolarizing population that showed a close "linear" relationship between "granularity" (90 degrees depolarization) and "lobularity" (90 degrees polarization). This atypical population was smaller than normal leukocytes and thus clearly different from the patterns associated with malaria infection. Abnormal depolarization patterns of M. perstans clearly do not reflect leukocyte-associated malaria hemozoin. It is possible however that the erythrocyte-lysing agent used to facilitate leukocyte analysis by the instrument may have caused microfilaria fragmentation and thus the distinctive "straight-line" features of the abnormal scatter plots

[Pneumococcal bacteremia in patients aged over 65 years. A study of 161 cases]. / Bacteriemia neumocócica en pacientes mayores de 65 años. Estudio de 161 casos.

BACKGROUND: Here we present the results from a prospective analysis of pneumococcal bacteremia in patients older than 65 years, highlighting its differences with regard to patients aged from 15 to 64 years. PATIENTS AND METHOD: We performed a prospective follow-up study (1988-1999) of patients older than 14 years whohad blood cultures positive for Streptococcus pneumoniae(S. pneumoniae). S. pneumoniae was identified using standard techniques. The antimicrobial sensitivity was analysed using the broth midrodilution technique and the serotype was studied in a reference laboratory. Statistical analyses were performed by means of the SPSS program, version 9.0. RESULTS: 161 out of 321 bacteremias occurred in 154 patients older than 65 years (86 males). Most frequent underlying diseases were chronic obstructive pulmonary disease (COPD), diabetes and neoplasia. Most patients had pneumonia, 29.7% with ilfiltrates involving at least two lung lobes. 9.4% of bacteremias were nosocomial. Among patients older than 65 years, 32.5% of strains were resistant to penicillin, compared to 21.7% in patients younger than 65 years(p = 0.034). Mortality in patients over 65 years was higher than in younger patients (25.3% vs 11.5%; p = 0.002) and mortality predictive factors in a multivariate analysis were age, nosocomial bacteremia and multi-lobe involvement (in patients with pneumonia). CONCLUSIONS: There is a high incidence of pneumococcal bacteremia in patients over 65 years of age. These patients have a higher rate of resistance to penicillin and a higher mortality rate than younger patients. Mortality predictive factors are age, nosocomial bacteremia and multi-lobe involvement.

Limited level of accuracy provided by available rapid diagnosis tests for malaria enhances the need for PCR-based reference laboratories.

The rise of imported malaria cases and the high fatality rate in Europe make the search for new and easy diagnostic methods necessary. Rapid diagnosis tests (RDTs) are, in part, developed to cover the lack of diagnosis experience. Unfortunately, our data suggest that the accuracy of RDTs is insufficient and could increase the number of incorrect malaria diagnoses.

The use of PCR in the diagnosis of hyper-reactive malarial splenomegaly (HMS).

Between August 1997 and September 1998, 14 cases of hyper-reactive malarial splenomegaly (HMS) were diagnosed in the Instituto de Salud Carlos III in Madrid, Spain. These cases, from Equatorial Guinea and Cameroon, were identified using the diagnostic criteria established by Y. M. Fakunle in 1981: gross splenomegaly; high levels of anti-malarial antibodies; IgM in serum at least two standard deviations above the local mean; and clinical and immunological response to antimalarial treatment. Although malarial parasites were only detected in the Giemsa-stained blood films of four of the cases, these four and four others were found to have the DNA of such parasites in their blood when tested using a method based on a semi-nested, multiplex PCR. These result indicate that malarial parasitaemias may be more prevalent in HMS than is usually recognized.

[Evaluation of different clinical samples and cell lines in the isolation of enterovirus in pediatric patients]. / Evaluación de diferentes muestras clínicas y líneas celulares en el aislamiento de Enterovirus en pacientes pediátricos.

BACKGROUND: To evaluate prospectively the efficacy of different clinical samples and cellular lines in the isolation of Enteroviruses from pediatric patients. METHODS: In the period july-1997 to july-1999 we analyze the samples of 102 pediatric patients (< 2 years old) with an unknown febrile etiology syndrome. After the process of decontamination the samples were inoculated in the MRC-5, Hep-2 and Vero cell lines by the shell-vial assay. After 2-3 days of incubation the monolayers were stained by a monoclonal antibody against VP1; then the strains were identified as Poliovirus, ECHO-virus or Coxsacke by specific antibodies. RESULTS: 96 clinical samples were studied (45 pharyngeal swabs, 28 feces, 13 cerebrospinal fluid, 5 bloods, 4 urines and one bronchoalveolar lavage). Enteroviruses were isolated in 48 patients (47%), corresponding to 60 clinical samples (62.5%). The enteroviruses were isolated in 75.5% of pharyngeal swabs, 71.4% of feces, 30.7% of cerebrospinal fluid, one blood (20%) and the bronchoalveolar lavage. 28 patients presented simultaneously both pharyngeal and rectal swabs; in this group the use of both samples permitted the isolation of Enteroviruses in 26 cases (92.8%). Of 60 Enteroviruses, 59 (98.3%) were isolated in the MRC-5 cell line, 23 (38.3%) in the Hep-2, and 14 (23.3%) in the Vero; the MRC-5 demonstrated a significant difference in the isolation of Enteroviruses compared with the other cell lines. The Enteroviruses were identified as 30 (50%) ECHO-virus, 15 (25%) as Poliovirus vaccine, 12 (20%) could not to be identified and in 3 cases (5%) no growth of virus was detected in the culture. The MRC-5 cell line was significatively superior to the other in the isolation of the different Enteroviruses. CONCLUSIONS: To obtain the maximum diagnostic efficacy in front a patient with a possible infection by Enteroviruses, is necessary to use different clinical samples, specially the more related to the infection focci (cerebrospinal fluid or blood). The MRC-5 cell line has been the most efficace in the isolation of Enteroviruses independently of the type of clinical sample or the viral genus. The shell-vial assay is a good method for the isolation and identification of Enteroviruses isolated from clinical samples.

Usefulness of seminested multiplex PCR in surveillance of imported malaria in Spain.

The use of a new PCR-based method for the diagnosis of malaria in the Spanish Malaria Reference Laboratory has promoted an increase in confirmed cases of malaria. From August 1997 to July 1998, a total of 192 whole-blood samples and 71 serum samples from 168 patients were received from the hospitals of the Spanish National Health System. Most of the patients came from west-central African countries (85%). This molecular method showed more sensitivity and specificity than microscopy, detecting 12.4% more positive samples than microscopy and 13% of mixed infections undetectable by Giemsa stain. Plasmodium falciparum was the main species detected, with 68% of the total positive malaria cases, followed by Plasmodium malariae (29%), Plasmodium vivax (14%), and Plasmodium ovale (7%), including mixed infections in all cases. This report consists of the first wide, centralized survey of malaria surveillance in Spain. The reference laboratory conducted the analysis of all imported cases in order to detect trends in acquisition. The use of a seminested multiplex PCR permitted confirmation of the origins of the infections and the Plasmodium species involved and confirmation of the effectiveness of drug treatments. This PCR also allowed the detection of the presence in Spain of primaquine-tolerant P. vivax strains from west-central Africa, as well as the detection of a P. falciparum infection induced by transfusion.

Induction of histamine release in parasitized individuals by somatic and cuticular antigens from Onchocerca volvulus.

The host immune response in onchocerciasis is believed to contribute to the clinical manifestations of infection. Mazzotti and chronic inflammatory reactions might be mediated by mechanisms involving specific IgE and reactivity of mast cells and basophils to the parasite antigens. In this report, we show that Onchocerca volvulus antigens are capable of inducing histamine release. Three types of extracts were prepared from the parasite: soluble total, surface, and cuticular collagen. Soluble extracts released histamine in all individuals with onchocerciasis at significantly higher levels (P < 0.05) than those found in endemic controls, but similar levels to those found in patients with mansonellosis. However, cuticular collagen induced significantly (P < 0.01) higher histamine release in patients with onchocerciasis than in those with mansonellosis. No reactivity against human type IV collagen was observed. Implications derived from the presence of sensitized basophils in the pathogenesis of onchocerciasis are discussed.

[Eosinophilic gastroenteritis caused by bee pollen sensitization]. / Gastroenteritis eosinofílica causada por sensibilización a polen de abeja.

A 34-year-old Spanish woman with a lifelong history of seasonal rhinoconjunctivitis and honey intolerance (pyrosis and abdominal pain) developed, 3 weeks after starting ingestion of bee pollen, astenia, anorexia, abdominal pain, diarrhoea, peripheral blood hypereosinophilia and elevated serum total IgE levels. A duodenal biopsy showed eosinophilic infiltration of the mucosal layer. Other causes of hypereosinophilia were not found. Repeated parasitological stool studies, as well as a duodenal aspirate showed negative results. Symptoms, hypereosinophilia and elevated IgE levels resolved after bee pollen ingestion was stopped. This is a typical case of eosinophilic gastroenteritis by ingestion of bee pollen in a woman with intolerance to honey bee, because the patient fulfilled the usual diagnostic criteria: gastrointestinal symptoms were present, eosinophilic infiltration of the digestive tract was demonstrated by biopsy, no eosinophilic infiltration of other organs was found and the presence of parasites was excluded. Honey intolerance and/or bee pollen administration should be considered as a cause of eosinophilic gastroenteritis.

Microsporidiosis in HIV-positive children in Madrid (Spain).

A prospective study was carried out to determine the prevalence rates of microsporidiosis and other enteroparasites in HIV-positive children in the Madrid area. HIV-positive pediatric patients from three hospitals were enrolled in the study. A total of 293 samples (158 stool and 127 urine) were collected from 83 children whose mean age was 6.3 years and had a mean CD4 count of 504.7/mm3 (range 1-2,220/mm3), 48 of whom suffered diarrhea at the time of the study. Microsporidia identification was investigated in stool and urine samples using Weber's chromotrope-based stain, IIF and PCR species-specific tests. Enteric parasites were identified in 32.5% of the children. Cryptosporidium sp. was the most common parasite encountered (14.4%), followed by Blastocytis sp. (9.6%) and Giardia duodenalis (8.4%). Microsporidia was only found in the stools of one child (1.2% of total and 2% of those with diarrhea) and Enterocytozoon bieneusi was demonstrated by PCR. The patient was 10 years old, presented non-chronic diarrhea and his CD4 count was 298/mm3. These data differ from those previously reported by us in HIV-positive adults (13.9%) in the same area, although this group showed more severely depressed CD4 lymphocyte counts than children. New epidemiological studies should be carried out to elucidate whether additional risk factors exist between these groups.

[Species identification of intestinal microsporidiosis in HIV-positive patients using the polymerase chain reaction]. / Identificación específica de microsporidiosis intestinales en pacientes HIV-positivos mediante la reacción en cadena de la polimerasa.

BACKGROUND: Microsporidia are opportunistic parasites which, due to their morphologic characteristics, continue presenting diagnostic problems. Species-specific identification of microsporidia has become important because of varying levels of response to albendazole, which is the only effective treatment for some kinds of intestinal microsporidiosis. Although these parasites cause up to 50% of otherwise unexplained chronic diarrhea in HIV-positive patients, the number of reported cases is still very scarce in our country when compared to the existing HIV-positive population. METHODS: Intestinal microsporidiosis in HIV-positive patients with diarrhea was investigated using the modified trichrome staining technique. Microsporidia species identification was done by indirect immunofluorescence (IIF) and polymerase chain reaction (PCR) with specific primers. RESULTS: Six new cases of intestinal microsporidiosis caused by Enterocytozoon bieneusi were diagnosed in Madrid (Spain). All patients were in an advanced state of the HIV infection and they presented CD4+ values equal or inferior to 100 x 10(6)/I. CONCLUSIONS: Due to the number of cases that are accumulating, microsporidia must be included among the enteropathogens responsible for chronic diarrhea in HIV-positive individuals in Spain. The PCR technique using specific primers is a suitable determinator of the microsporidia species implicated in this intestinal pathology.

[Diagnosis of 4 cases of intestinal microsporidiosis in AIDS patients]. / Diagnóstico de 4 casos de microsporidiosis intestinal en enfermos de SIDA.

BACKGROUND: Most of the latest clinical and epidemiologic studies indicate that microsporidiosis and above all Enterocytozoon bieneusi cause approximately 7-50% of otherwise unexplained diarrhea in HIV-infected patients. Four cases of intestinal microsporidiosis in AIDS-patients are reported. METHODS: Detection of spores of microsporidiosis in stool samples from HIV-infected patients was performed by the following techniques: 1) light microscopy, using Kokoskin's modification of the Weber trichrome stain; 2) electron microscopy, for confirmation (only available in one case), and 3) indirect immunofluorescence assay (IIF), performed on all positive samples. Rabbit polyclonal antibodies anti-Encephalitozoon cuniculi, anti-E. hellem and anti-E. (Septata) intestinalis were used to eliminate the possibilities of presence of microsporidiosis other than E. bieneusi. RESULTS: Detection of spores of microsporidiosis by trichrome stain was positive in all samples from the 4 patients. By elimination, IIF confirmed the presence of Enterocytozoon bieneusi in all 4 cases. E. bieneusi was identified by electron microscopy in the only case available. Three patients had CD4+ lymphocyte counts < 100 x 10(6)/l and one < 500 x 10(6)/l. All had AIDS and three of them were homosexual. Two patients remained positive for microsporidiosis for almost 5 months. CONCLUSIONS: Despite the very few cases described in Spain, these preliminary data lead us to believe that intestinal microsporidiosis should be considered as an possible cause of diarrhea in our country also, and mainly in acute immunodeficiency patients.