Prevalencia de parásitos intestinales en niños preescolares del municipio Angostura del Orinoco, estado Bolívar, Venezuela. 2016-2018 / Prevalence of intestinal parasites in preschool children from the Angostura del Orinoco municipality, Bolívar state, Venezuela. 2016-2018

El objetivo de la presente investigación fue determinar la prevalencia de parásitos intestinales en niños preescolares usuarios de 12 Centros de Educación Inicial Simoncito (CEIS) del municipio Angostura del Orinoco, estado Bolívar, Venezuela. Se estudiaron las heces de 515 niños de ambos géneros (2 a 5 años), mediante la técnica de sedimentación espontánea. La prevalencia de enteroparásitos fue de 39,4% (n=203). No hubo diferencias estadísticamente significativas (χ2 = 1,59 g.l.: 2 p> 0,05) respecto a la edad, pero si según el género (p<0,05), resultando los niños varones más afectados con 46,9%. Se identificaron 11 taxones de enteroparásitos, destacando el cromista Blastocystis spp. con 27,4% (n=141) y el protozoario Giardia intestinalis con 13,2% (n=68). Se encontró una baja prevalencia de helmintos, donde Ascaris lumbricoides fue el más común con 1,6% (n=8). De los 203 niños parasitados, el 70,9% (n=144) estaba monoparasitado y 29,1% (n=59) poliparasitado. Los principales parásitos asociados fueron Blastocystis spp. (89,8%), G. intestinalis (44,1%) y Endolimax nana (35,3%). En conclusión, se determinó una elevada prevalencia de parásitos intestinales en la población estudiada, por lo que estas infecciones continúan siendo un problema de salud pública en niños de Ciudad Bolívar, Venezuela

The objective of the present investigation was to determine the prevalence of intestinal parasites in preschool children users of 12 Simoncito Initial Education Centers (CEIS) of the Angostura del Orinoco municipality, Bolívar state, Venezuela. The feces of 515 children of both genders (2 to 5 years old) were studied using the spontaneous sedimentation technique. The prevalence of enteroparasites was 39.4% (n = 203). There were no statistically significant differences (χ2 = 1.59 d.f .: 2 p> 0.05) with respect to age but if according to gender (p <0.05), because the most affected were male child with 46.9%. Eleven taxa of enteroparasites were identified, highlighting the chromist Blastocystis spp. (27.4%) and the protozoan Giardia intestinalis (13.2%). A low prevalence of helminths was found, where Ascaris lumbricoides was the most common with 1.6%. Of the 203 parasitized children, 70.9% (n = 144) were monoparasitized and 29.1% (n = 59) polyparasitized. The main associated parasites were Blastocystis spp. (89.8%), G. intestinalis (44.1%) and Endolimax nana (35.3%). In conclusion, a high prevalence of intestinal parasites was determined in the population studied, so these infections continue to be a public health problem in children from Ciudad Bolívar, Venezuela

The effect of water source and soil supplementation on parasite contamination in organic vegetable gardens / O efeito da fonte de água e suplementação de solo na contaminação parasitária em hortas orgânicas

Abstract The objective of this study was to determine factors associated with vegetable contamination with zoonotic protozoan. Samples of water, soil and vegetables were collected from July/2014 to May/2016, totaling 83 samples, 21 properties of Londrina region, Paraná, Brazil. DNA amplification of Toxoplasma gondii, Cryptosporidium spp. and Giardia intestinalis in the samples was conducted using polymerase chain reaction (PCR). The PCR results were positive for T. gondii in 12.9% (8/62), Cryptosporidium spp. in 11.3% (7/62) and G. intestinalis in 25.8% (16/62) of the samples. DNA sequencing identified C. parvum in five samples and G. intestinalis Assemblage E in three. The statistical associations demonstrated greater probability of positive samples for T. gondii and for at least one of the three protozoa when the source of irrigation water was the river; a greater chance of positive samples for Cryptosporidium spp. when deer were present on the property; and a smaller chance of positive samples for at least one of the three etiologic agents when soil was supplemented with limestone. The results expose some critical contamination points, providing support for training farmers on good management practices during the production process.

Resumo O trabalho teve como objetivo determinar os fatores associados à contaminação de vegetais por protozoários zoonóticos. Amostras de água, solo e vegetais foram coletadas de julho/2014 a maio/2016, totalizando 83 amostras de 21 propriedades da região de Londrina, Paraná, Brasil. A amplificação de fragmentos de DNA de T. gondii, Cryptosporidium spp. e Giardia intestinalis foi realizada por meio da reação em cadeia da polimerase (PCR). Os resultados da PCR foram positivos para T. gondii em 12,9% (8/62), Cryptosporidium spp. em 11,3% (7/62) e G. intestinalis. em 25,8% (16/62) das amostras. O sequenciamento de DNA identificou C. parvum em cinco amostras e G. intestinalis, Assemblage E em três amostras. As associações estatísticas evidenciaram maior probabilidade de amostras serem positivas para T. gondii ou para pelo menos um dos três protozoários quando a fonte de água de irrigação era o rio; uma maior chance de amostras positivas para Cryptosporidium spp. quando havia cervos na propriedade; e uma menor chance das amostras serem positivas para pelo menos um dos três agentes etiológicos quando o solo era suplementado com calcário. Os resultados expõem alguns pontos críticos de contaminação, fornecendo suporte para capacitar os agricultores em boas práticas de gestão durante o processo de produção.

Prevalence of Intestinal Parasites and Associated Risk Factors for Infection among Rural Communities of Chachoengsao Province, Thailand

Intestinal parasitic infections remain a major public health problem in many parts of Thailand, particularly in rural areas. This study aimed to determine the prevalence of intestinal parasitic infections and associated risk factors among the people living in Huai Sai sub-district, Bang Khla district, Chachoengsao Province, central Thailand. A cross-sectional survey was carried out from June 2017 to August 2017 which included a total of 224 participants. Stool samples were examined using a simple direct smear and formalin ethyl acetate concentration technique. Association between risk factors and intestinal parasitic infections was assessed using multivariate logistic regression analysis. The overall prevalence of intestinal parasitic infections was 16.1%. Soil-transmitted helminth infections (14.3%) were more common than protozoan infections (1.8%). The most common intestinal parasites were hookworms (6.7%) followed by Strongyloides stercoralis, (5.0%), Ascaris lumbricoides (1.3%) and Trichuris trichiura (1.3%). Entamoeba histolytica/dispar (1.0%), Giardia intestinalis (0.4%), and Blastocystis hominis (0.4%) were the protozoans identified. A high prevalence of infections was found in male participants of ≥40 years who raised dogs in the households and did not wear boots while working fields. Multivariate analysis showed a significant association of intestinal parasitic infections with gender with the adjusted odds ratio (OR) of 2.4 and 95% confidence interval (CI) of 1.1–5.2 (P=0.020). The results showed a high prevalence of soil-transmitted helminth infections among adults in rural communities which were particularly apparent regarding the skin-penetrating species of nematodes. A greater focus on intervention is required by improving sanitation and personal hygiene to prevent the spread of intestinal parasitic infections.

Parasitosis y síndrome de intestino irritable / Parasitosis and irritable bowel syndrome

El síndrome de intestino irritable (SII) es un trastorno funcional digestivo de etiología multifactorial. En su fisiopatología se describen diversos factores, tanto biológicos, como psicológicos y ambientales, que afectan el estado de activación de células inmunes en la mucosa intestinal. Entre los factores ambientales se incluye la presencia de alguna parasitosis intestinal. El síndrome de intestino irritable post-infeccioso (SII-PI) es reconocido como un subgrupo de estos trastornos, cuya aparición de los síntomas es posterior a una infección intestinal provocada por agentes microbianos. A pesar de que en Chile hay pocos estudios respecto a la relación entre SII y parasitosis intestinal, se ha descrito la existencia de una asociación positiva entre SII e infecciones por Blastocistis hominis, uno de los parásitos prevalentes en Chile. En otros países, se ha descrito además una relación entre SII, amebiasis y giardiasis. Por la alta prevalencia de parasitosis en nuestro país, existe la necesidad de ampliar los estudios para clarificar la fuerza de la asociación entre parasitosis y SII.

Irritable bowel syndrome (IBS) is a functional disorder of the gastrointestinal tract characterised by multi-factorial aetiology. In IBS physiopathology are involved diverse factors between them biological, psychosocial, and environmental components which affect the immune activation status of gut mucosa. Among these factors is recognized the intestinal parasitosis. Post-infection IBS (PI-IBS) is recognised as a subgroup of functional disorders whose symptoms onset appear after a symptomatic intestinal infection caused by microbial agents. There are few studies regarding of relationship between IBS and intestinal parasitosis in Chile. However, is has been well described a positive association between IBS and Blastocystis hominis infections, one of prevalent parasites in Chile. In other countries, is also described a relationship between IBS and amebiasis and giardiasis. Both, characterized by a common mode of transmission through water as well as contaminated food. Because the high prevalence of parasitosis in our country it is necessary to expand the association studies to clarify the strength of the parasites ethiology in IBS.

Relación clínico-epidemiológica de giardiasis en niños de 0-12 años que asisten a núcleos de atención primaria. municipio francisco linares alcántara, estado aragua, venezuela / Clinical-epidemiologic relationship of giardiasis in children 0-12 years attending primary care nucleus.municipality francisco linares alcantara, aragua, venezuela

Giardiasis is an intestinal parasitism with high prevalence worldwide, which most often affects children. In Venezuela is a public health problem due to the occurrence of various risk factors, such as pollution of sources of drinking water, poor health education and low socioeconomic status. A descriptive cross-sectional study in children attending Primary Care Nucleus (NAP) of the municipality Francisco Linares Alcantara, during the first quarter of 2013. Stool samples were a nalyzed by direct coproparasitological methods with physiological Saline, Lugol and Faust. In order to know the symptoms and risk factors associated with a clinical epidemiological survey was applied to each participant. The sample consisted of 180 children, of which 47.2% had pathogenic parasites, 14.4% non-pathogenic and 38.3% were not parasitized. The prevalence of G. intestinalis was 11.1%. No relationship between sex and age group was found. Association between diarrhea, flatulence and abdominal distention and presence of G. intestinalis was found. Risk factors associated with the presence of the parasite were: poor housing, treatment of drinking water and inadequate hygiene. We conclude that G. intestinalis is a common pathogen in children who come to the NAP of the municipality and its presence is associated with poor housing, inadequate treatment of drinking water and inadequate hygiene.

La giardiasis es una parasitosis intestinal con alta prevalencia a nivel mundial, que afecta con mayor frecuencia a niños. En Venezuela constituye un problema de salud pública, debido a la concurrencia de diversos factores de riesgo, como contaminación de las fuentes de agua de consumo humano, la deficiente educación sanitaria y nivel socioeconómico bajo. Se realizó un estudio descriptivo y transversal en niños y niñas que acudieron a los Núcleos de Atención Primaria (NAP) del municipio Francisco Linares Alcántara, durante el primer trimestre del año 2013. Las muestras de heces fueron analizadas mediante métodos coproparasitológicos directos con solución Salina Fisiológica, Lugol y Faust. Para conocer la sintomatología y los factores de riesgo asociados, se aplicó una encuesta clínico epidemiológica a cada participante. La muestra estuvo conformada por un total de 180 niños, de los cuales 47,2% presentaban parásitos patógenos, 14,4% no patógenos y 38,3% no estaban parasitados. La prevalencia de Giardia intestinalis fue de 11,1%. No se encontró relación entre sexo y grupo etario. Se halló asociación entre la diarrea, flatulencia y distensión abdominal y presencia de G. intestinalis. Los factores de riesgo asociados a la presencia de dicho parásito fueron: precariedad de la vivienda, tratamiento del agua de consumo humano y hábitos higiénicos inadecuados. Se concluye que G. intestinalis es un patógeno frecuente en los niños que acuden a los NAP del municipio y su presencia está asociada a la precariedad de la vivienda, tratamiento inadecuado del agua de consumo humano y hábitos higiénicos inadecuados.

Determinación del ensamblaje genético de aislados axénicos colombianos de Giardia intestinalis / Genetic assemblages of axenic Colombian cultures of Giardia intestinalis

Resumen Objetivo: Genotipificar muestras de Giardia aisladas de pacientes de tres regiones geográficas de Colombia. Materiales y métodos: Se examinaron 22 aislados de G. intestinalis, los primeros descritos cultivados axénicamente de origen colombiano por medio del análisis del polimorfismo de longitud de fragmentos de restricción (RFLP) del gen glutamato deshidrogenasa (gdh). Resultados y conclusión: El patrón del RFLP mostró que todos los aislados pertenecían al ensamblaje A, específicamente al AI; resultado confirmado por secuenciación. Con este estudio se pretende contribuir al conocimiento de los genotipos circulantes de Giardia intestinalis en Colombia.

Abstract Objective: To genotype samples of G. intestinalis isolated from patients of three Colombian regions. Materials and methods: 22 isolates of Giardia (all of them established as axenic cultures) were examined by analyzing restriction fragment length polymorphism (RFLP) of the glutamate dehydrogenase gene (gdh). Results and conclusion: The RFLP patterns showed that all of the Giardia isolates belonged to the assembly A, specifically to AI. This result was confirmed by sequencing. This study aims to contribute to the knowledge of circulating genotypes of Giardia intestinalis in Colombia.

Analysis of Intestinal Parasitic Infections in a Tertiary Care Hospital of Ahmadabad.

Background & Objectives: Microscopic evidence of Intestinal parasitic infections (IPIs) becomes evident only when the parasites are heavily populated in the intestine. Immunological tests can diagnose IPIs even at earlier stage of a disease but the tests are costly, not easily available and lots of research is yet to be done in that field. So even today we have to rely upon microscopic evidence only & microscopically detected infections may be a tip of iceberg only. The purpose of this study was to determine the distribution of Intestinal parasitic infections in patients attending a tertiary care hospital. Material and Methods: A total of 847 stool sampleswere examined macroscopically & microscopically for the presence of parasite eggs, larva, cysts and trophozoites. Results: Out of 847 stool samples, parasites were detected from 72 (8.5%) samples. The distribution of intestinal parasites was as follows: Entamoeba histolytica (44%), Giardia intestinalis (30%), Trichomonas intestinalis (10%), Ascaris lumbricoides (6%), Hymenolepis nana (4%), Strongyloides stercoralis (2%), Ancylostoma duodenale (2%), and Isospora belli (1%). Conclusion: It is concluded that protozoal infections are more common in our region than helminthic infections. Constant surveillance of the infected patients, their treatment as well as improving sanitary condition will help to prevent the spread of IPIs.

Comparative analysis of enzyme-linked immunosorbent assay and direct microscopy for the diagnosis of Giardia intestinalis in fecal samples.

Context: Giardiasis is one of the most common nonviral infections causing diarrheal illness worldwide. In this prospective cross-sectional study, we evaluated the RIDASCREEN® Giardia kit for detection of Giardia intestinalis in stool samples and compared the results with direct microscopy. Materials and methods: A total of 360 fecal samples were collected. They were then processed by wet fi lm, iodine preparation and an enzyme-linked immunosorbent assay (ELISA) kit to determine the presence of Giardia trophozoites and cysts. Statistical analysis was performed by sensitivity, specifi city, positive predictive value, negative predictive value and diagnostic accuracy. Results and Conclusion: Of the 360 cases, 17.2% samples were positive for Giardia by direct microscopy and 23.6% were found to be positive by ELISA (sensitivity ∼97%), but specifi city was ∼92% only. Because of less specifi city, we need to perform ELISA in congruence with direct microscopy, etc. Further studies need to be performed on a larger sample size using other molecular tests in order to get more accurate estimations

Cryptosporidium spp., Giardia intestinalis, and Enterocytozoon bieneusi in Captive Non-Human Primates in Qinling Mountains

Non-human primates (NHPs) are confirmed as reservoirs of Cryptosporidium spp., Giardia intestinalis, and Enterocytozoon bieneusi. In this study, 197 fresh fecal samples from 8 NHP species in Qinling Mountains, northwestern China, were collected and examined using multilocus sequence typing (MLST) method. The results showed that 35 (17.8%) samples were positive for tested parasites, including Cryptosporidium spp. (3.0%), G. intestinalis (2.0%), and E. bieneusi (12.7%). Cryptosporidium spp. were detected in 6 fecal samples of Macaca mulatta, and were identified as C. parvum (n=1) and C. andersoni (n=5). Subtyping analysis showed Cryptosporidium spp. belonged to the C. andersoni MLST subtype (A4, A4, A4, and A1) and C. parvum 60 kDa glycoprotein (gp60) subtype IId A15G2R1. G. intestinalis assemblage E was detected in 3 M. mulatta and 1 Saimiri sciureus. Intra-variations were observed at the triose phosphate isomerase (tpi), beta giardin (bg), and glutamate dehydrogenase (gdh) loci, with 3, 1, and 2 new subtypes found in respective locus. E. bieneusi was found in Cercopithecus neglectus (25.0%), Papio hamadrayas (16.7%), M. mulatta (16.3%), S. sciureus (10%), and Rhinopithecus roxellana (9.5%), with 5 ribosomal internal transcribed spacer (ITS) genotypes: 2 known genotypes (D and BEB6) and 3 novel genotypes (MH, XH, and BSH). These findings indicated the presence of zoonotic potential of Cryptosporidium spp. and E. bieneusi in NHPs in Qinling Mountains. This is the first report of C. andersoni in NHPs. The present study provided basic information for control of cryptosporidiosis, giardiasis, and microsporidiosis in human and animals in this area.

Molecular Detection of Giardia intestinalis from Stray Dogs in Animal Shelters of Gyeongsangbuk-do (Province) and Daejeon, Korea

Giardia is a major public health concern and considered as reemerging in industrialized countries. The present study investigated the prevalence of giardiosis in 202 sheltered dogs using PCR. The infection rate was 33.2% (67/202); Gyeongsangbuk-do and Daejeon showed 25.7% (39/152, P or =1 year of age which showed only 27.4% positive rate, 61.8% (P<0.001) of the total samples collected from young animals (<1 year of age) were positive for G. intestinalis. A significantly higher prevalence in symptomatic dogs (60.8%, P<0.0001) was observed than in asymptomatic dogs (23.8%). Furthermore, the analysis of nucleotide sequences of the samples revealed that G. intestinalis Assemblages A and C were found in the feces of dogs from Gyeongsangbuk-do and Daejeon. Since G. intestinalis Assemblage A has been known to infect humans, our results suggest that dogs can act as an important reservoir of giardiosis in Korea. Hence, hygienic management should be given to prevent possible transmission to humans.

Giardíase / Giardiasis

Este artigo tem como objetivo realizar uma revisão sobre aspectos clínicos, diagnóstico, tratamento e profilaxia da giardíase. A Giardia lamblia é um parasita intestinal de ampla distribuição mundial e elevada prevalência em países com baixo desenvolvimento socioeconômico. A giardíase apresenta um amplo espectro clínico e as principais manifestações incluem diarreia e má absorção intestinal. Está associada a síndrome do intestino irritável e a redução da função cognitiva em crianças. O exame parasitológico de fezes é um método de baixo custo tradicionalmente usado para o diagnóstico de giardíase. Outros métodos incluem os testes de detecção de antígeno em amostras de fezes, a biópsia e a análise do fluido duodenal. O tratamento da giardíase deve ser adaptado para cada grupo de pacientes, considerando posologia, efeitos adversos, interações medicamentosas, gestação e prevalência de outras parasitoses. As principais medidas para o controle da giardíase são tratamento adequado da água, lavagem dos alimentos e boa higiene pessoal...

Preliminary study of the enzyme ubiquitin carboxylterminal hydrolase 14 (UBP6) in Giardia intestinalis: structural bioinformatic analysis and transcriptional profile during encystation / Estudio preliminar de la enzima ubiquitina carboxiterminal hidrolasa 14 (UBP6) en Giardia intestinalis: análisis bioinformático de su estructura y perfil transcripcional durante la enquistación / Estudo preliminar da enzima ubiquitina carboxi-terminal hidrolasa 14 (UBP6) em Giardia intestinalis: análise bioinformática estrutural e perfil transcricional durante encistamento

This paper presents a combined approach with two aims. The first is to analyze the reported sequence of the enzyme ubiquitin carboxyl-terminal hydrolase 14 of Giardia intestinalis (UBP6) through computational methods to find components related with its hypothetical function. The second is to determine if the protein-coding gene is expressed in G. intestinalis and, if such is the case, also determine its transcription pattern along the life cycle of the parasite. It was established that the protein belongs to the family of Cys-dependent deubiquitinases and more specifically to ubiquitin specific proteases (USPs). Moreover, the catalytic center with the complete triad as well as typical features of the USP motif were also identified. Since the computational findings suggest that the enzyme could be functional, reverse transcription coupled to PCR was used as a first approach to establish if in fact the coding gene is expressed in the parasite. Interestingly, it was found not only that the gene is expressed, but also that there is a transcription variation along the life cycle of the parasite. These two findings are the starting point for further studies since they tentatively suggest that this enzyme could be involved in the protein turnover that occurs during parasite encystation. Although preliminary, this study is the first report concerning the study of a specific deubiquitinating enzyme in the parasite G. intestinalis.

En este trabajo se presenta una estrategia combinada que buscaba, primero, analizar por métodos computacionales la secuencia de la enzima ubiquitina carboxilo-terminal hidrolasa 14 de Giardia intestinalis (UBP6) reportada para buscar componentes relacionados con su función hipotética y segundo, determinar si el gen que codifica para la proteína se expresa en G. intestinalis y si lo hace, cómo es su patrón de transcripción a lo largo del ciclo de vida del parásito. Se encontró que la proteína pertenece a la familia de deubiquitinasas dependientes de cisteína y más específicamente a las proteasas específicas para ubiquitina (USPs por ubiquitin specific proteases). También se identificaron el centro catalítico con la triada completa así como características típicas del motivo USP. Teniendo en cuenta que los resultados computacionales sugieren que la enzima puede ser funcional, se usó la técnica de transcripción reversa acoplada a PCR como un primer acercamiento para establecer si el gen codificante se expresa en el parásito. De manera interesante, se determinó no solo que el gen se expresa sino que existe una variación de su transcripción a lo largo del ciclo de vida del parásito. Estos hallazgos son el punto de partida para posteriores estudios ya que sugieren de manera preliminar que esta enzima podría estar involucrada en el recambio de proteínas que ocurre en el parásito durante el proceso de enquistación. Aunque preliminar, este estudio es el primer reporte acerca de una enzima deubiquitinadora específica en el parásito G. intestinalis.

Este artigo apresenta uma abordagem combinada com dois objetivos. A primeira é analisar a sequência informou da enzima ubiquitina carboxil-terminal hidrolase 14 de Giardia intestinalis (UBP6) através de métodos computacionais para encontrar os componentes relacionados com a sua função hipotética. A segunda é para determinar se o gene de codificação da proteína é expressa em G. intestinalis e, se for o caso, também determinar o seu padrão de transcrição ao longo do ciclo de vida do parasita. Foi estabelecido que a proteína pertence à família de deubiquitinases Cys-dependentes e mais especificamente para proteases específicas de ubiquitina (USPs por ubiquitin specific proteases). Além disso, o centro catalítico com a tríade completo, bem como as características típicas do motivo USP também foram identificados. Uma vez que os resultados computacionais sugerem que a enzima poderia ser funcional, a transcrição reversa acoplada a PCR foi utilizado como uma primeira abordagem para determinar se, de facto, o gene codificante é expressa no parasita. Curiosamente, verificou-se não só que o gene é expresso, mas também que há uma variação de transcrição ao longo do ciclo de vida do parasita. Estes dois elementos são o ponto de partida para estudos posteriores, uma vez que tentativas sugerem que esta enzima pode estar envolvida no refill de proteínas que ocorre durante o parasita encistamento. Embora preliminares, este estudo é o primeiro relatório relativo ao estudo de uma enzima deubiquitinadora específica no parasita intestinalis.

Molecular characterisation of Giardia intestinalis assemblages from human isolates at a tertiary care centre of India.

Purpose: The aim of the study was to determine the genetic heterogeneity of Giardia intestinalis isolates detected in stool samples of the study population using polymerase chain reaction assay and restriction fragment length polymorphism. We also tried to correlate the association/differences between the clinical symptomatology and infection by different assemblages (genotypes) of G. intestinalis. Materials and Methods: This cross-sectional study was conducted from April 2008 to June 2010. A total of 40 adults (n = 40) and 42 children (n = 42) below the age of 12 years with the clinical suspicion of giardiasis and with the onset of one or more of the following fi ve symptoms, i.e., loose stool, nausea, weight loss, fatigue and foul smelling faeces and confi rmed laboratory diagnosis of giardiasis at least once during the current episode of diarrhoea were included in this study. Results: Of the 82 patients (males 66) enrolled in the study, 70 (85%) presented with diarrhoea (56 males) and 12 (15%) without diarrhoea (10 males). Out of 70 diarrheic patients, 61 (87%) had chronic diarrhoea, 8 (11.5%) had acute diarrhoea and 1 (1.5%) had persistent diarrhoea. Of the total patients, 63 (77%) were clinically assessed and were apparently immunocompetent, whereas, 19 (23%) immunocompromised patients had different underlying conditions besides giardiasis. Genotyping identifi ed all 82 (100%) isolates as assemblage B. Conclusion: We found that assemblage B of G. intestinalis presents with all kinds of clinical features ranging from asymptomatic carriage to acute, persistent or chronic diarrhoea.

Atualidades sobre giardíase / Updates on giardiasis

A giardíase é uma parasitose intestinal amplamente distribuída pelo mundo, com alta prevalência em países em desenvolvimento, inclusive o Brasil. Embora seja uma infecção com bom prognóstico, pode apresentar gravidade em pessoas com desnutrição, fibrose cística ou algumas imunodeficiências. Este artigo tem como objetivo atualizar os principais aspectos da giardíase.

Giardiasis is an intestinal parasite widely distributed around the globe with high prevalence in developing countries, including Brazil. Although it is an infection with a good prognosis may present serious in people with malnutrition, cystic fibrosis or certain immunodeficiencies. The aim of this paper is to perform an update on key aspects of giardiasis.

High occurrence of giardiasis in children living on a 'landless farm workers' settlement in Araras, São Paulo, Brazil / Alta ocorrência de giardíase em crianças do Assentamento Sem Terra, Araras, São Paulo, Brasil

Enteric parasitosis remains an important public health problem in many areas around the world including in Brazil, and it is frequently associated with poverty and lack of sanitation facilities. Research carried out over the course of a year revealed that 96.6% (28/29) of children randomly selected from a 'landless farm workers' settlement in Araras, São Paulo, aged 4 - 15 years, presented Giardia intestinalis cysts. After referral to the neighborhood Health Office, all the children received tinidazole, given as a single dose of 50 mg/kg and 12 months later, new fecal samples were collected and analyzed. Despite the low adherence to the study, a high percentage (64.3% - 9/14) of the children remained positive for the parasite. This study showed a high positivity of giardiasis in child residents of the settlement, even after treatment; adults were not sensitized to the study and did not collected and/or deliver children fecal samples. The precarious living conditions are consistent with a high susceptibility to parasitic diseases, suggesting that the treatment of the infected individuals without identifying and eradicating the means of contamination is simply a palliative measure.

Enteroparasitoses continuam a ser um importante problema de saúde pública em muitas áreas ao redor do mundo, bem como no Brasil, e está frequentemente associada com a pobreza e à falta de saneamento básico. Pesquisa realizada em um ano revelou que 96,6% (28/29) das crianças com idades entre quatro e 15 anos, recrutadas aleatoriamente no Assentamento Sem Terra em Araras, São Paulo, apresentaram cistos de Giardia intestinalis. Após o encaminhamento ao Posto de Saúde do bairro, todos receberam tinidazol, dose única de 50 mg/kg. Após 12 meses, novas amostras de fezes foram coletadas e analisadas. Apesar da baixa adesão ao estudo, um percentual elevado (64,3% - 9/14) de crianças permaneceu positivo para o protozoário. Este estudo mostrou alta positividade de giardíase nas crianças moradoras do assentamento, mesmo após o tratamento; indivíduos adultos não se mostraram sensibilizados com o estudo e não coletaram e/ou entregaram amostras fecais dos filhos; e o tratamento dos indivíduos infectados, sem identificação e erradicação das formas de contaminação, só funciona como medida paliativa.

Producción y purificación de anticuerpos aviares (IgYs) a partir de cuerpos de inclusión de una proteína recombinante central en el metabolismo del NAD+ / Production and purification of avian antibodies (IgYs) from inclusion bodies of a recombinant protein central in NAD+ metabolisim / Produção e purificação de anticorpos aviários (IgYs) empregando os corpos de inclusão de uma proteína recombinante importante para o metabolismo do NAD+

El uso de gallinas para la producción de anticuerpos policlonales, reduce la intervención animal, obteniendo además una gran cantidad de anticuerpos. Las aves presentan mayor distancia filogenética entre sus antígenos y los de mamíferos ofreciendo altos porcentajes de anticuerpos específicos. La producción de anticuerpos requiere antígenos en cantidades considerables, por ello es creciente el uso de proteínas recombinantes para tales fines. No obstante, la obtención de proteínas en sistemas heterólogos conduce frecuentemente a la precipitación en agregados insolubles de limitada utilidad (cuerpos de inclusión). Este trabajo presenta una metodología para la producción de anticuerpos policlonales (IgYs) empleando cuerpos de inclusión (CI) como antígeno. Los CI purificados e inoculados corresponden a la Nicotinamida / Nicotinato Mononucleotido Adenililtranferasa (His-GiNMNAT) de Giardia intestinalis expresada en Escherichia coli. La purificación del antígeno se llevó a cabo mediante solubilización y renaturalización. Los anticuerpos se purificaron de la yema de huevo de gallinas imunizadas mediante dilución en agua, seguida de precipitación con sulfato de amonio al 60% y afinidad tiofilica. Los anticuerpos fueron evaluados mediante inmunoblot empleando la proteína His-GiNMNAT. De una yema de huevo se obtuvieron 14,4 mg de IgYs, con alta pureza y con un reconocimiento de hasta 15ng de His-GiNMNAT. Se mejoró la especificidad de los IgYs mediante una purificación adicional por afinidad al antígeno, lo cual permitiría su empleo para el reconocimiento de la proteína del parásito.

In contrast with other animal models, the use of hens for polyclonal antibodies production not only reduces animal intervention, but also increases the quantity of the obtained immunoglobulins. The phylogenetic distance between birds and mammals, leads to more specific avian antibodies than their mammalian counterparts. Since a large amount of antigen is required for avian antibody production, the use of recombinant proteins for this procedure has been growing faster over the last years. Nevertheless, recombinant protein production through heterologous systems, frequently leads to the formation of insoluble and useless aggregates (inclusion bodies, IC). This article presents a strategy to produce avian polyclonal antibodies (IgYs) from IC. In order to obtain the antigen, the Giardia intestinalis nicotinamide mononucleotide adenylyltransferase recombinant protein (His-GiNMNAT) was expressed in Escherichia coli. The His-GiNMNAT protein was purified through IC solubilization and re-folding. The purified protein was use to immunize hens. The antibodies were purified from egg yolk by water dilution, followed by ammonium sulfate precipitation and thiophilic affinity chromatography. Specificity of the purified antibodies was tested against the His-GiNMNAT protein through Western blot essays. In terms of yield, 14.4 mg of highly pure IgYs were obtained from one egg yolk; these antibodies were able to detect 15 ng of antigen. IgYs specificity was improved by means of antigen affinity purification, allowing its implementation for endogenous detection of GiNMNAT protein in G. intestinalis.

A utilização de galinhas para a produção de anticorpos policlonais, diminui a intervenção sobre o animal e permite a geração de grandes quantidades de anticorpos. As aves têm maior distância filogenética entre os seus antígenos em comparação com os mamíferos, oferecendo altos porcentagens de anticorpos específicos. A produção de anticorpos requerem quantidades consideráveis de antigénio, por conseguinte, é comum a utilização de proteínas recombinantes para esta finalidade. No entanto, a produção de proteínas em sistemas heterólogos muitas vezes leva à precipitação em agregados insolúveis de utilidade limitada (corpos de inclusão). Este trabalho apresenta uma metodologia para a produção de anticorpos policlonais, utilizando proteína recombinante a partir de corpos de inclusão. O antígeno utilizado foi a proteína Nicotinamida / Nicotinato Mononucleótido Adenililtranferasa de Giardia intestinalis gerada em Escherichia coli (His-GiNMNAT). A purificação do antigénio foi feita por solubilização e renaturalização. Os anticorpos foram purificados a partir da gema de ovo de galinhas imunizadas pelo método de diluição em água, seguido de precipitação com sulfato de amónio (60%) e afinidade tiofilica. Os anticorpos foram avaliados por imunoblot utilizando a proteína His-GiNMNAT. De uma gema de ovo foram obtidos 14,4 mg de IgYs com pureza elevada. Estes anticorpos podem reconhecer até 15 ng de His-GiNMNAT. A especificidade dos IgYs foi reforçada por outra purificação por afinidade pelo antigénio. Isto irá permitir a sua utilização para o reconhecimento da proteína do parasita.

Stool sample storage conditions for the preservation of Giardia intestinalis DNA

Stool is chemically complex and the extraction of DNA from stool samples is extremely difficult. Haemoglobin breakdown products, such as bilirubin, bile acids and mineral ions, that are present in the stool samples, can inhibit DNA amplification and cause molecular assays to produce false-negative results. Therefore, stool storage conditions are highly important for the diagnosis of intestinal parasites and other microorganisms through molecular approaches. In the current study, stool samples that were positive for Giardia intestinalis were collected from five different patients. Each sample was stored using one out of six different storage conditions [room temperature (RT), +4ºC, -20ºC, 70% alcohol, 10% formaldehyde or 2.5% potassium dichromate] for DNA extraction procedures at one, two, three and four weeks. A modified QIAamp Stool Mini Kit procedure was used to isolate the DNA from stored samples. After DNA isolation, polymerase chain reaction (PCR) amplification was performed using primers that target the β-giardin gene. A G. intestinalis-specific 384 bp band was obtained from all of the cyst-containing stool samples that were stored at RT, +4ºC and -20ºC and in 70% alcohol and 2.5% potassium dichromate; however, this band was not produced by samples that had been stored in 10% formaldehyde. Moreover, for the stool samples containing trophozoites, the same G. intestinalis-specific band was only obtained from the samples that were stored in 2.5% potassium dichromate for up to one month. As a result, it appears evident that the most suitable storage condition for stool samples to permit the isolation of G. intestinalis DNA is in 2.5% potassium dichromate; under these conditions, stool samples may be stored for one month.

Prevalencia de giardia intestinalis en habitantes del barrio la macarena, Ciudad Bolívar, Venezuela / Prevalence of Giardia Intestinalis in population of la macarena community, Ciudad Bolívar, Venezuela

Introducción: La giardiasis es una parasitosis intestinal producida por el protozoario Giardia intestinalis. Aunque de trascendencia clínica importante, por lo general se le presta poca atención o no se piensa en ella al momento de hacer el diagnóstico. Se realizó un estudio con el objetivo de determinar la prevalencia de G. intestinalis en habitantes del Barrio La Macarena de Ciudad Bolívar, estado Bolívar. Materiales y métodos: En abril de 2008 fueron evaluados 136 habitantes de la comunidad. Una muestra de heces de cada individuo fue analizada mediante las técnicas de examen directo, Kato y sedimentación espontánea. Resultados: La prevalencia de parásitos intestinales fue de 58,1 % (79/136). Se diagnosticaron 9 especies de parásitos y/o comensales, con un predominio de los protozoarios. Giardia intestinalis fue el segundo parasito más prevalente (19,9 %), superado sólo por Blastocystis sp. (59 %). En estos 27 casos de giardiasis, no hubo diferencias con relación al sexo (p > 0,05), pero con relación a la edad ya que los menores de 15 años resultaron más afectados. El 96 % de los casos de infección por G. intestinalis estaba asociado a otros protozoarios intestinales. Conclusión: Se determinó una elevada prevalencia de G. intestinalis en habitantes del Barrio La Macarena de Ciudad Bolívar.

Introduction: Giardiasis is an intestinal parasitism caused by the protozoan Giardia intestinalis. Although it has clinical transcendence very important, generally little attention is bean paid or is not taught of at the time of making the diagnosis. To determine of G. intestinalis prevalence in people from La Macarena community in Ciudad Bolívar, Bolívar state, a study carried out. Material and methods: In April of 2008 136 inhabitants were evaluated. A sample of stool of every people was analyzed by direct examination, Kato and spontaneous sedimentation methods. Results: The prevalence of intestinal parasitism was 58.1 % (79/136). Nine species of parasites or commensals were diagnosed, with a predominance of protozoans. Giardia intestinalis was the second parasite most prevalent (19.9 %), it was only dominated by Blastocystis sp. (59.6 %). In these 27 cases of giardiasis, there were no differences with regard to sex (p > 0.05) in infected people, but a with regard to age, the 15 years under people resulted more affected. 96.3 % of cases of G. intestinalis infections was associated to others intestinal protozoans. Conclusion: We determined a high prevalence of G. intestinalis in population of La Macarena community, Ciudad Bolívar.

Prevalencia de Giardia intestinalis y predominio de genotipos zoonóticos en ovinos y bovinos de traspatio de cinco estados de la República Mexicana / Prevalence of Giardia intestinalis and zoonotic genotype predominance in small scale sheep and cattle farms in five states of the Mexican Republic

The aim of this paper was to discover the prevalence and assemblages of Giardia intestinalis, harbored in sheep and cows on familiar farms from five states of the Mexican Republic. Stool samples from 265 sheep and 174 cows were analyzed by centrifugation and flotation in zinc sulfate to search for cysts and ova. The samples with Giardia cysts were processed in a Sheather solution in order to isolate them. Afterwards, cultures were established in TYI-S-33, each one of which was the Giardia DNA source. The DNA was obtained and used as a template to amplify a fragment of the glutamate dehydrogenase (gdh) enzyme. The 430 bp amplicons were restricted with Nla IV and Rsa I in order to identify the restriction fragments length polymorphisms (RFLP's) patterns. From the cyst analysis, Giardia cysts in nine cows (5.1%) and 30 sheep (11.3%) were found. Then 10 axenic cultures (5 from sheep and 5 from cows) were set up. From the RFLP's pattern it was found that one cow had assemblage (AI), another two had a mixture of assemblages (AI + BIII) and the other two had (E + BIII). In sheep, it was found that two sheep had assemblage (AI) and the other three had a mixture of assemblages (AI + BIII). This is the first report in which zoonotic assemblages (A-I and BIII) predominance in ruminants from five states of Mexico have been demonstrated. Therefore, it is necessary to carry out further studies aimed at discovering other Giardia genotypes and transmission patterns between animals and humans in Mexico.

Con el fin de determinar la frecuencia y genotipos de Giardia intestinalis en ovinos y bovinos de traspatio de algunos estados de la República Mexicana, en este trabajo se colectaron heces de 265 ovinos y 174 bovinos, para la búsqueda de Giardia mediante coproparasitoscópicos (CPS) de concentración flotación. De las muestras fecales que resultaron positivas se obtuvieron los quistes por el método de Sheather. Los quistes se desenquistaron in vitro y los trofozoítos se mantuvieron en cultivo TYI-S-33 axénico. El ADN de los trofozoítos se obtuvo mediante extracciones fenólicas y se amplificó un segmento de ≈ 430 pb del gen de la enzima glutamato deshidrogenasa (gdh) por medio de la reacción en cadena de la ADN polimerasa (PCR), el producto se restringió con las enzimas Nla IV y Rsa I y se obtuvieron los polimorfismos de los fragmentos de restricción (RFLP). En los CPS se encontró a Giardia en nueve bovinos (5.1%) y 30 ovinos (11.3%). Se establecieron 10 cultivos axénicos (5 de bovinos y 5 de ovinos). En un bovino se encontró el genotipo (AI), dos tuvieron mezcla de los genotipos (AI + BIII) y los otros dos fueron (E + BIII). Un ovino fue genotipo (AI) y tres tuvieron mezcla de los genotipos (AI + BIII). Éste es el primer informe que presenta predominio de genotipos zoonóticos (AI y BIII) en ovinos y bovinos de México. Es necesario investigar los genotipos de Giardia y patrones de transmisión entre animales y humanos en México.

Detecção de Giardia intestinalis pelo teste de impressão duodenal e infecção por Helicobacter pylori em crianças e adolescentes com sintomas digestivos / Giardia intestinalis detected by biopsy imprint and Helicobacter pylori infection in dyspeptic children and adolescents

Objetivo: avaliar a relação entre a infecção por helicobacter pylori e a presença de giardia intestinalis, diagnosticada através do teste de impressão da mucosa duodenal em pacientes com dor abdominal. Material e métodos: foram avaliados prospectivamente 33 pacientes consecutivos com queixa de dor abdominal e que realizaram exame parasitológico de fezes; durante a endoscopia digestiva alta foram coletadas biópsias de duodeno para o teste de impressão e análise microscópica, e de mucosa do antro para pesquisa do H. pylori (teste rápido da urease e histologia). Resultados: o teste da impressão da mucosa duodenal foi positiva para giardia intestinalis em 9/33 pacientes (27,2%) e o H. pylori foi detectado em 21/33 (63,6%) pacientes, sendo 4/9 (44,4%) entre os pacientes infestados e 17/24 (70,8%) entre os não infestados por Giardia intestinalis. O exame parasitológico de fezes foi positivo em apenas 1/9 (11%) pacientes com o teste de impressão positivo; outros 3 pacientes apresentaram: ascaris lumbricoides, entamoeba coli e endolimax nana e a biópsia duodenal para giardia intestinalis foi positiva em 3/8 (37,5%). Conclusão: nossos resultados sugerem que não há associação entre parasitoses e infecção por Helicobacter pylori, sendo o teste de impressão da mucosa duodenal útil no diagnóstico da infestação por Giardia intestinalis durante investigação de pacientes com sintomas dispépticos submetidos à endoscopia digestiva. No entanto, são necessários mais estudos com maior amostragem.

Objective: to evaluate the relationship between helicobacter pylori infection and giardia intestinalis infestation, diagnosed by duodenal imprint test, in patients with abdominal pain. Material and methods: thirty three consecutive patients were prospectively evaluated by esophagogastroduodenoscopy and stool examination for ova and parasites. Duodenal biopsy was collected for imprint test and histological examination. H. pylori was diagnosed by histological examination of gastric antrum and rapid urease test. Results: duodenal imprint test was positive for giardia intestinalis in 9/33 patients (27.2%) and H. ylori was detected in 21/33 (63.6%) patients, 4/9 (44.4%) giardia intestinalis positive patients and 17/24 (70.8%) giardia intestinalis negative patients. Stool examination was positive in 1/9 (11%) patient with a positive duodenal imprint test, and another 3 patients presented ascaris lumbricoides, entamoeba coli e endolimax nana. Duodenal histology was positive for giardia intestinalis in 3/8 (37,5%). Conclusion: our results suggested that there is no association between giardiasis and Helicobacter pylori infection. Duodenal imprint test was useful to diagnose giardia intestinalis infestation in dyspeptic patients evaluated by esophagogastroduodenoscopy. Nevertheless, more studies with greater sampling are necessary.