Prevalence of Entamoeba histolytica and other enteral parasitic diseases in the metropolitan region of Belo Horizonte, Brazil. A cross-sectional study.

BACKGROUND: Enteral parasitic diseases are a public health problem in nations with low economic development and in settings with poor sanitation. Amebiasis is the second most frequent form of parasitosis, with a high burden of disease. Knowledge of the prevalence of enteroparasitoses in a given region is useful for planning clinical decision-making. Thus, the aim of this study was to estimate the prevalence of enteral parasitic diseases, especially amebiasis, through analysis on stool samples from public and private laboratories in a metropolitan area in southeastern Brazil. DESIGN AND SETTING: Cross-sectional study conducted in the metropolitan region of Belo Horizonte, Brazil. METHODS: We evaluated 6,289 fecal samples from one private and one public laboratory. The samples were concentrated by means of spontaneous sedimentation, and those that were positive for Entamoeba histolytica or Entamoeba dispar in optical microscopy analyses were processed to obtain deoxyribonucleic acid, with subsequent identification through the polymerase chain reaction. RESULTS: Among the stool samples, 942 (15.0%) had parasitic infections; 73 (1.2%) of these were helminthic infections and 847 (13.5%) were protozoan infections, caused mainly by Escherichia coli (6.0%), Endolimax nana (5.2%) and Giardia lamblia (1.2%). Infections due to Entamoeba histolytica or Entamoeba dispar occurred in 36 samples (0.6%) and the polymerase chain reaction revealed five (13.9%) as Entamoeba histolytica. CONCLUSION: The prevalence of enteral parasitic diseases is high in the metropolitan region of Belo Horizonte, although amebiasis may not be a problem.

Prevalence of Entamoeba histolytica and other enteral parasitic diseases in the metropolitan region of Belo Horizonte, Brazil. A cross-sectional study

ABSTRACT BACKGROUND: Enteral parasitic diseases are a public health problem in nations with low economic development and in settings with poor sanitation. Amebiasis is the second most frequent form of parasitosis, with a high burden of disease. Knowledge of the prevalence of enteroparasitoses in a given region is useful for planning clinical decision-making. Thus, the aim of this study was to estimate the prevalence of enteral parasitic diseases, especially amebiasis, through analysis on stool samples from public and private laboratories in a metropolitan area in southeastern Brazil. DESIGN AND SETTING: Cross-sectional study conducted in the metropolitan region of Belo Horizonte, Brazil. METHODS: We evaluated 6,289 fecal samples from one private and one public laboratory. The samples were concentrated by means of spontaneous sedimentation, and those that were positive for Entamoeba histolytica or Entamoeba dispar in optical microscopy analyses were processed to obtain deoxyribonucleic acid, with subsequent identification through the polymerase chain reaction. RESULTS: Among the stool samples, 942 (15.0%) had parasitic infections; 73 (1.2%) of these were helminthic infections and 847 (13.5%) were protozoan infections, caused mainly by Escherichia coli (6.0%), Endolimax nana (5.2%) and Giardia lamblia (1.2%). Infections due to Entamoeba histolytica or Entamoeba dispar occurred in 36 samples (0.6%) and the polymerase chain reaction revealed five (13.9%) as Entamoeba histolytica. CONCLUSION: The prevalence of enteral parasitic diseases is high in the metropolitan region of Belo Horizonte, although amebiasis may not be a problem.

Evaluation of diagnostic methods for the detection of intestinal schistosomiasis in endemic areas with low parasite loads: Saline gradient, Helmintex, Kato-Katz and rapid urine test.

BACKGROUND: In some tropical countries, such as Brazil, schistosomiasis control programs have led to a significant reduction in the prevalence and parasite burden of endemic populations. In this setting, the Kato-Katz technique, as the standard diagnostic method for the diagnosis of Schistosoma mansoni infections, which involves the analysis of two slides from one fecal sample, loses its sensitivity. As a result, a significant number of infected individuals are not detected. The objective of this study was to perform extensive parasitological testing of up to three fecal samples and include a rapid urine test (POC-CCA) in a moderate prevalence area in Northern Minas Gerais, Brazil, and evaluate the performance of each test separately and in combination. METHODS AND FINDINGS: A total of 254 individuals were examined with variants of the standard Kato-Katz technique (up to18 Kato-Katz slides prepared from three fecal samples), a modified Helmintex (30 g of feces), the saline gradient (500 mg of feces), and the POC-CCA methods. We established a reference standard taking into consideration all the positive results in any of the parasitological exams. Evaluation of the parasite burden by two Kato-Katz slides confirmed that most of the individuals harbored a light infection. When additional slides and different parasitological methods were included, the estimated prevalence rose 2.3 times, from 20.4% to 45.9%. The best sensitivity was obtained with the Helmintex method (84%). All parasitological methods readily detected a high or moderate intensity of infection; however, all lost their high sensitivity in the case of low or very low intensity infections. The overall sensitivity of POC-CCA (64.9%) was similar to the six Kato-Katz slides from three fecal samples. However, POC-CCA showed low concordance (κ = 0.34) when compared with the reference standard. CONCLUSIONS: The recommended Kato-Katz method largely underestimated the prevalence of S. mansoni infection. Because the best performance was achieved with a modified Helmintex method, this technique might serve as a more precise reference standard. An extended number of Kato-Katz slides in combination with other parasitological methods or with POC-CCA was able to detect more than 80% of egg-positive individuals; however, the rapid urine test (POC-CCA) produced a considerable percentage of false positive results.

Hematological profile in natural progression of giardiasis: kinetics of experimental infection in gerbils.

INTRODUCTION: The clinical manifestations of giardiasis and its impact are harmful to children, and may cause deficits in their physical and cognitive development. The pathogenic mechanisms are usually unknown and the available reports can be controversial. METHODOLOGY: The present study aimed to know, for the first time, the evolution of the hematological profile of the gerbils, experimentally infected with Giardia lamblia, up to the infection's resolution. Hematological variables have been tested. RESULTS: White blood cells have not presented meaningful alterations during the course of the infection. A significant reduction in the number of red blood cells (p = 0.021), in the concentration of hemoglobin (p = 0.029) and in the value of the hematocrit (p = 0.016) has been observed, starting from the second week of infection, ratifying an anemia related to giardiasis. Reduction in the level of serum iron starting from the third week of infection, despite not being significant, could suggest the participation of iron in the anemia. However, the weight of the animals was kept and the hematimetric parameters started to return to the basic values after the parasitological cure without iron reposition. CONCLUSIONS: The outcomes found suggest the idea that not only malabsorption but also other mechanisms such as chronic inflammation may be implicated in iron deficiency anemia in giardiasis and may explain how asymptomatic patients may have anemia without malabsorption. In this context, considering the highlighting character of the anemia in our study, we believe that anemia should be investigated in children with giardiasis. And in the cases of anemia without a definite etiology, giardiasis should also be investigated.

Differentially expressed genes of virulent and nonvirulent Entamoeba histolytica strains identified by suppression subtractive hybridization.

Entamoeba histolytica is a parasite which presents capacity to degrade tissues and therefore has a pathogenic behavior. As this behavior is not shown by all strains, there have been several studies investigating molecular basis of the cytotoxicity process. Using the suppression subtractive hybridization (SSH) technique, differential gene expressions of two E. histolytica strains, one virulent (EGG) and one nonvirulent (452), have been analyzed with the purpose of isolating genes which may be involved with amoebic virulence. Nine cDNA fragments presenting high homology with E. histolytica previously sequenced genes were subtracted. Of these, four genes were confirmed by RT-PCR. Two coding for hypothetical proteins, one for a cysteine-rich protein, expressed only in the virulent strain, EGG and another one, coding for grainin 2 protein, exclusive from 452 strain. This study provided new insight into the proteins differences in the virulent and nonvirulent E. histolytica strains. We believe that further studies with these proteins may prove association of them with tissue damage, providing new perceptions to improve treatment or diagnosis of the invasive disease.

High prevalence of enteroparasitosis in urban slums of Belo Horizonte-Brazil. Presence of enteroparasites as a risk factor in the family group.

The present study evaluates the prevalence of enteroparasitosis in the urban slums of Belo Horizonte, Brazil and the risk of transmitting enteroparasites to the family members of infected individuals. Stool samples were collected and examined at clinical laboratories near each slum. Individuals were identified and classified as positive for parasitosis (IP(+)), and individuals with negative stool tests were classified as negative for parasitosis (IP(-)) and enrolled as control patients. We collected samples from 594 patients, of which 20·2% and 79·8% were classified as IP(+) and IP(-), respectively. In addition, 744 family members (FIPs) effectively participated in the study by providing fecal samples. In total, 1338 participants were evaluated. Of these, 34·6% were tested positive for parasitosis. Blastocystis was the most prevalent parasite, infecting 22·4% of individuals. Among FIPs, the overall prevalence was 46·1%. Of these, 50·6% and 44·7% were classified as FIPs(+) and FIPs(-), respectively. These results showed that IP(+) did not impact the prevalence of infection within the studied communities, not constituting index cases of specific risk behaviors, suggesting that, in fact, these communities are exposed to similar oral-fecal routes of contamination.

Prevalence of intestinal parasitism and associated symptomatology among hemodialysis patients.

Intestinal parasites are an important cause of morbidity and mortality. Immunocompromised individuals may develop more severe forms of these infections. Taking into account the immunity impairment in patients suffering from chronic renal failure (CRF), we will determine the prevalence and associated symptoms of intestinal parasites in these patients. Controls without CRF were used for comparison. Stool samples were collected and processed for microscopic identification of parasites using the Formalin-ether concentration method. For Cryptosporidium diagnosis, the ELISA technique was used. One hundred and ten fecal samples from hemodialysis patients were analyzed, as well as 86 from a community group used as control group. A result of 51.6% of intestinal parasites was observed in hemodialysis patients and 61.6% in the control group. Cryptosporidium and Blastocystis were the most common infections in patients with CRF (26.4% and 24.5%, respectively). Blastocystis was the most common infection in the control group (41.9%), however no individual was found positive for Cryptosporidium. Among the CRF patients, 73.6% were symptomatic, 54.3% of these tested positive for at least one parasite, in contrast to 44.8% in asymptomatic patients (p = 0.38). The most common symptoms in this group were flatulence (36.4%), asthenia (30.0%) and weight loss (30.0%). In the control group, 91.9% were symptomatic, 60.8% of these tested positive for at least one parasite, in contrast to 71.4% in asymptomatic patients (p = 0.703). A significant difference between the two groups was observed with regard to symptoms, with bloating, postprandial fullness, and abdominal pain being more frequent in the control group than in the hemodialysis group (all p < 0.05). Comparing symptomatic with asymptomatic, there was no association in either group between symptoms or the prevalence of parasitic infection, nor with the type of parasite or with multiple parasitic infections. Patients with chronic renal failure are frequent targets for renal transplantation, which as well as the inherent immunological impairment of the disease itself, results in immunosuppression by medication. For this reason, carriers of intestinal parasites with pathogenic potential can develop serious clinical complications influencing the success of transplantation. This fact, coupled with the high prevalence of intestinal parasites and the dissociation between symptoms and infection in CRF patients, suggests that the stool test should be incorporated in routine propedeutics. Furthermore, preventive measures for the acquisition of parasites through the fecal-oral contamination route should be introduced.

Prevalence of ittestinal parasitism and associated symptomatology among hemodialysis patients / Prevalência de parasitismo intestinal e sintomatologia associada em pacientes hemodialíticos

Intestinal parasites are an important cause of morbidity and mortality. Immunocompromised individuals may develop more severe forms of these infections. Taking into account the immunity impairment in patients suffering from chronic renal failure (CRF), we will determine the prevalence and associated symptoms of intestinal parasites in these patients. Controls without CRF were used for comparison. Stool samples were collected and processed for microscopic identification of parasites using the Formalin-ether concentration method. For Cryptosporidium diagnosis, the ELISA technique was used. One hundred and ten fecal samples from hemodialysis patients were analyzed, as well as 86 from a community group used as control group. A result of 51.6% of intestinal parasites was observed in hemodialysis patients and 61.6% in the control group. Cryptosporidium and Blastocystis were the most common infections in patients with CRF (26.4% and 24.5%, respectively). Blastocystis was the most common infection in the control group (41.9%), however no individual was found positive for Cryptosporidium. Among the CRF patients, 73.6% were symptomatic, 54.3% of these tested positive for at least one parasite, in contrast to 44.8% in asymptomatic patients (p = 0.38). The most common symptoms in this group were flatulence (36.4%), asthenia (30.0%) and weight loss (30.0%). In the control group, 91.9% were symptomatic, 60.8% of these tested positive for at least one parasite, in contrast to 71.4% in asymptomatic patients (p = 0.703). A significant difference between the two groups was observed with regard to symptoms, with bloating, postprandial fullness, and abdominal pain being more frequent in the control group than in the hemodialysis group (all p < 0.05). Comparing symptomatic with asymptomatic, there was no association in either group between symptoms or the prevalence of parasitic infection, nor with the type of parasite or with multiple parasitic infections. Patients with chronic renal failure are frequent targets for renal transplantation, which as well as the inherent immunological impairment of the disease itself, results in immunosuppression by medication. For this reason, carriers of intestinal parasites with pathogenic potential can develop serious clinical complications influencing the success of transplantation. This fact, coupled with the high prevalence of intestinal parasites and the dissociation between symptoms and infection in CRF patients, suggests that the stool test should be incorporated in routine propedeutics. Furthermore, preventive measures for the acquisition of parasites through the fecal-oral contamination route should be introduced.

Doenças parasitárias infectam grande número de indivíduos em todo o mundo. Manifestações clínicas mais severas podem se apresentar em pacientes imunocomprometidos. Considerando o importante comprometimento imunológico observado em pacientes com insuficiência renal crônica (IRC), foi determinada a prevalência e sintomas associados a parasitoses intestinais nesses pacientes em comparação a controles saudáveis. Foram coletadas amostras fecais de cada participante e processadas para identificação microscópica dos parasitas pelo método de concentração por formol-éter. Foi utilizada a técnica de ELISA para identificar coproantígenos de Cryptosporidium. Foram analisadas 110 amostras fecais de pacientes em hemodiálise e 86 de um grupo controle comunitário. Cryptosporidium e Blastocystis foram as infecções mais freqüentes nos pacientes em hemodiálise (26,4% e 24,5%, respectivamente). Blastocystis foi a infecção mais freqüente no grupo controle (41,9%), entretanto nenhum indivíduo positivo para Cryptosporidium foi identificado. Considerando os pacientes com IRC, 73,6% eram sintomáticos, sendo 54,3% positivos para algum parasita, contra 44,8% nos assintomáticos (p = 0,38). Os sintomas mais frequentes neste grupo foram flatulência (36,4%), adinamia (30,0%) e perda de peso (30,0%). No grupo controle, 91,9% eram sintomáticos, sendo 60,8% positivos para algum parasita, contra 71,4% nos assintomáticos (p = 0,703). Em relação aos sintomas, houve diferença significativa entre os dois grupos, sendo que flatulência, plenitude pós-prandial, e dor abdominal foram mais freqüentes no grupo controle que nos pacientes em hemodiálise (todos p < 0,05). Comparando-se sintomáticos com assintomáticos, não houve associação entre a sintomatologia e a prevalência de parasitose, nem com o tipo de parasita, e nem com o poliparasitismo, nos dois grupos. Considerando que pacientes com IRC são frequentes alvos de transplante renal, resultando em imunossupressão por medicamentos, que é somada à deficiência imunológica inerente à própria doença. Os portadores de parasitas intestinais com potencial patogênico podem desenvolver sérias complicações clínicas que influenciam o sucesso do transplante. Este fato, aliado a alta prevalência de parasitas intestinais e dissociação entre os sintomas e infecção nesses pacientes, sugerem a incorporação do exame de fezes na propedêutica de rotina dos mesmos, juntamente com medidas preventivas para a aquisição de parasitas com rota de contaminação fecal-oral.

Effects of metronidazole analogues on Giardia lamblia: experimental infection and cell organization.

The chemotherapeutic agents used for the treatment of giardiasis are often associated with adverse side effects and are refractory cases, due to the development of resistant parasites. Therefore the search for new drugs is required. We have previously reported the giardicidal effects of metronidazole (MTZ) and its analogues (MTZ-Ms, MTZ-Br, MTZ-N(3), and MTZ-I) on the trophozoites of Giardia lamblia. Now we evaluated the activity of some giardicidal MTZ analogues in experimental infections in gerbils and its effects on the morphology and ultrastructural organization of Giardia. The giardicidal activity in experimental infections showed ED(50) values significantly lower for MTZ-I and MTZ-Br when compared to MTZ. Transmission electron microscopy was employed to approach the mechanism(s) of action of MTZ analogues upon the protozoan. MTZ analogues were more active than MTZ in changing significantly the morphology and ultrastructure of the parasite. The analogues affected parasite cell vesicle trafficking, autophagy, and triggered differentiation into cysts. These results coupled with the excellent giardicidal activity and lower toxicity demonstrate that these nitroimidazole derivates may be important therapeutic alternatives for combating giardiasis. In addition, our results suggest a therapeutic advantage in obtaining synthetic metronidazole analogues for screening of activities against other infectious agents.