Enteropathogens associated with diarrheal disease in infants of poor urban areas of Porto Velho, Rondônia: a preliminary study

One hundred and thirty cases of diarrhea and 43 age-matched controls, 0 to 5 years old, were studied in a pediatric outpatient unit from a poor peri urban area of Porto Velho, Rondônia. Eighty percent of diarrheal cases were observed in the groups under 2 years of age. Rotavirus (19.2%) was the most frequent enteropathogen associated with diarrhea, followed by Shigella flexneri (6.15%) and S. sonnei (1.5%) and Salmonella sp. (6.9%). Four cases of E. coli enterotoxigenic infections (3.1%), E. coli enteropathogenic (EPEC)(2.3%) one case of E. coli enteroinvasive infection (0.8%) and one case of Yersinia enterocolitica (0.8%) were also identified. Mixed infections were frequent, associating rotavirus, EPEC and Salmonella sp. with Entamoeba histolytica and Giardia lamblia.

A simple modification of the Baermann method for diagnosis of strongyloidiasis

The diagnosis of Strongyloides stercoralis infections is routinely made by microscopic observation of larvae in stool samples, a low sensitivity method, or by other, most effective methods, such as the Baermann or agar culture plate methods. We propose in this paper a practical modification of Baermann method. One hundred and six stool samples from alcoholic patients were analyzed using the direct smear test, agar culture plate method, the standard Baermann method, and its proposed modification. For this modification the funnel used in the original version of the method is substituted by a test tube with a rubber stopper, perforated to allow insertion of a pipette tip. The tube with a fecal suspension is inverted over another tube containing 6 ml of saline solution and incubated at 37°C for at least 2 h. The saline solution from the second tube is centrifuged and the pellet is observed microscopically. Larva of S. stercoralis were detected in six samples (5.7%) by the two versions of the Baermann method. Five samples were positive using the agar culture plate method, and only in two samples the larva were observed using direct microscopic observation of fecal smears. Cysts of Endolimax nana and Entamoeba histolytica/dyspar were also detected in the modification of Baermann method. Data obtained by the modified Baermann method suggest that this methodology may helps concentrate larvae of S. stercoralis as efficiently as the original method.

PAP1 [poly(A) polymerase 1] homozygosity and hyperadenylation are major determinants of increased mRNA stability of CDR1 in azole-resistant clinical isolates of Candida albicans.

Using genetically matched azole-susceptible (AS) and azole-resistant (AR) clinical isolates of Candida albicans, we recently demonstrated that CDR1 overexpression in AR isolates is due to its enhanced transcriptional activation and mRNA stability. This study examines the molecular mechanisms underlying enhanced CDR1 mRNA stability in AR isolates. Mapping of the 3' untranslated region (3' UTR) of CDR1 revealed that it was rich in adenylate/uridylate (AU) elements, possessed heterogeneous polyadenylation sites, and had putative consensus sequences for RNA-binding proteins. Swapping of heterologous and chimeric lacZ-CDR1 3' UTR transcriptional reporter fusion constructs did not alter the reporter activity in AS and AR isolates, indicating that cis-acting sequences within the CDR1 3' UTR itself are not sufficient to confer the observed differential mRNA decay. Interestingly, the poly(A) tail of the CDR1 mRNA of AR isolates was approximately 35-50 % hyperadenylated as compared with AS isolates. C. albicans poly(A) polymerase (PAP1), responsible for mRNA adenylation, resides on chromosome 5 in close proximity to the mating type-like (MTL) locus. Two different PAP1 alleles, PAP1-a/PAP1-alpha, were recovered from AS (MTL-a/MTL-alpha), while a single type of PAP1 allele (PAP1-alpha) was recovered from AR isolates (MTL-alpha/MTL-alpha). Among the heterozygous deletions of PAP1-a (Deltapap1-a/PAP1-alpha) and PAP1-alpha (PAP1-a/Deltapap1-alpha), only the former led to relatively enhanced drug resistance, to polyadenylation and to transcript stability of CDR1 in the AS isolate. This suggests a dominant negative role of PAP1-a in CDR1 transcript polyadenylation and stability. Taken together, our study provides the first evidence, to our knowledge, that loss of heterozygosity at the PAP1 locus is linked to hyperadenylation and subsequent increased stability of CDR1 transcripts, thus contributing to enhanced drug resistance.

Parasitism in Kansas in the 1800s: a glimpse to the past through the analysis of grave sediments from Meadowlark cemetery

During the excavations of the XIX century Meadowlark cemetery (Manhattan, Kansas, US), samples of sediments were taken from around five skeletons, and analyzed to detect intestinal parasites. No helminth eggs were found, but immunological ELISA tests for Entamoeba histolytica were positive in three samples. The immunological techniques have been successfully used in paleoparasitology to detect protozoan infections. Amoebiasis could have been a severe disease in the past, especially where poor sanitary conditions prevailed, and there is evidence that this cemetery may have been used in a situation where poor sanitary conditions may have prevailed. The presence of this protozoan in US during the late XIX century gives information on the health of the population and provides additional data on the parasite's evolution since its appearance in the New World.

A comparative study of the TF-Test®, Kato-Katz, Hoffman-Pons-Janer, Willis and Baermann-Moraes coprologic methods for the detection of human parasitosis

This study compares the diagnostic accuracy of the TF-Test® (TFT) for human parasitosis with results obtained using the traditional Kato-Katz (KK), Hoffman-Pons-Janer (HPJ), Willis and Baermann-Moraes (BM) techniques. Overall, four stool samples were taken from each individual; three alternate-day TFT stool samples and another sample that was collected in a universal container. Stool samples were taken from 331 inhabitants of the community of Quilombola Santa Cruz. The gold standard (GS) for protozoa detection was defined as the combined results for TFT, HPJ and Willis coproscopic techniques; for helminth detection, GS was defined as the combined results for all five coproscopic techniques (TFT, KK, HPJ, Willis and BM). The positivity rate of each method was compared using the McNemar test. While the TFT exhibited similar positivity rates to the GS for Entamoeba histolytica/dispar (82.4%) and Giardia duodenalis (90%), HPJ and Willis techniques exhibited significantly lower positivity rates for these protozoa. All tests exhibited significantly lower positivity rates compared with GS for the diagnosis of helminths. The KK technique had the highest positivity rate for diagnosing Schistosoma mansoni (74.6%), while the TFT had the highest positivity rates for Ascaris lumbricoides (58.1%) and hookworm (75%); HPJ technique had the highest positivity rate for Strongyloides stercoralis (50%). Although a combination of tests is the most accurate method for the diagnosis of enteral parasites, the TFT reliably estimates the prevalence of protozoa and selected helminths, such as A. lumbricoides and hookworm. Further studies are needed to evaluate the detection accuracy of the TFT in samples with varying numbers of parasites.

Amoeba/amoebal symbiont genetic transfers: lessons from giant virus neighbours.

Free-living amoebae serve as hosts for a variety of amoebae-resisting microorganisms, including giant viruses and certain bacteria. The latter include symbiotic bacteria as well as bacteria exhibiting a pathogenic phenotype towards amoebae. Amoebae-resisting bacteria have been shown to be widespread in water and to use the amoebae as a reservoir, a replication niche, a protective armour as well as a training ground to select virulence traits allowing survival in the face of microbicidal effects of macrophages, the first line of defense against invading pathogens. More importantly, amoebae play a significant role as a melting pot for genetic exchanges. These ecological and evolutionary roles of amoebae might also be at play for giant viruses and knowledge derived from the study of amoebae-resisting bacteria is useful for the study and understanding of interactions between amoebae and giant viruses. This is especially important since some genes have spread in all domains of life and the exponential availability of eukaryotic genomes and metagenomic sequences will allow researchers to explore these genetic exchanges in a more comprehensive way, thus completely changing our perception of the evolutionary history of organisms. Thus, a large part of this review is dedicated to report current known gene exchanges between the different amoebae-resisting organisms and between amoebae and the internalized bacteria.

Ocorrência de Enteroparasitas mais Frequentes entre Pacientes Atendidos no Hospital Dr. Agostinho Neto, Praia, Santiago

A qualidade em saúde, sua prevenção e manutenção são os principais problemas enfrentados nos países em desenvolvimento. De um modo geral, as informações sobre a ocorrência de enteroparasitas intestinais em Cabo Verde são escassas ou mesmo nulas para determinadas localidades ou mesmo ilhas. Considerando que as parasitoses apresentam variações inter-regionais, dependendo de condições sanitárias, educacionais, económicas, sociais, índice de aglomeração da população, condições de uso e contaminação do solo, da água e alimentos; e da capacidade de evolução das larvas e ovos de helmintos e de cistos de protozoários em cada um desses ambientes, procurou-se neste trabalho detalhar as características morfológicas, ciclo biológico e em especial alguns aspectos epidemiológicos de enteropasitos mais comumente registados. Os registos se reportaram a uma colecta retrospectiva de dados referentes aos meses de Julho a Setembro/2007, que foram gentilmente cedidos pelo Hospital Dr. Agostinho Neto e o Instituto Nacional de Estatística. As variáveis registadas durante as observações foram: mês, sexo, idade, residência, serviço, sintomas e infecção, e alguns referentes às condições de saneamento básico. Os resultados demonstraram que foram atendidos, com queixas relacionadas ao problema, principalmente indivíduos do sexo feminino, com idade de 25±19 anos, apresentando dores abdominais, e uma taxa de infecção por enteroparasitos de 27%. Os agentes mais frequentemente identificados durante o diagnóstico foram Giardia e Entamoeba coli, contudo também foram identificados a Entamoeba histolytica, a Endolimax nana e Heminolepis nana, em ordem de freqüência respectivamente, (fig. 17). Ficou também demonstrado que o depósito dos resíduos, sólidos e líquidos, foi realizado com maior frequencia ao redor da casa. A presença de água canalizada da rede pública, de chafariz e de fossas sépticas também foi registada, contudo, os resultados das análises descritivas demonstraram desvios muito elevados, (tabela 1). Os factores determinantes da taxa de infecção registada se relacionaram com a espécie ou gênero parasitário, a latitude e a idade do abordado, identificando-se também a presença de fossa séptica, rede de esgotos e água canalizada da rede pública como factores de dependência. O “depósito de resíduos ao redor da casa” e o “uso de água de nascente” influenciaram de forma parcial os resultados. Com base nesses resultados, indica-se que aproximadamente 40% tiveram amostras negativas para enteroparasitoses, estando este resultado relacionado ao local de moradia onde a deposição de resíduos foi menor, mesmo sendo registada a falta de rede de esgotos, água canalizada da rede pública e fossa séptica (fig.23). Aproximadamente 34% dos amostrados apresentaram índice de infecção baixo, se relacionando com a presença da deposição de lixo ao redor da casa, em detrimento do trabalho de criação de condições para instalação de água canalizada, fossa séptica e rede de esgotos, (fig.21) e finalmente que aproximadamente 24% dos amostrados apresentaram índice de infecção alto, este relacionado com a falta de água canalizada, de fossa séptica, de rede de esgotos, e o uso de água de nascente (fig.22).

Salacia crassifolia (Celastraceae): CHEMICAL CONSTITUENTS AND ANTIMICROBIAL ACTIVITY

The phytochemical study of hexane extract from leaves of Salacia crassifolia resulted in the isolation of 3β-palmitoxy-urs-12-ene, 3-oxofriedelane, 3β-hydroxyfriedelane, 3-oxo-28-hydroxyfriedelane, 3-oxo-29-hydroxyfriedelane, 28,29-dihydroxyfriedelan-3-one, 3,4-seco-friedelan-3-oic acid, 3β-hydroxy-olean-9(11):12-diene and the mixture of α-amirin and β-amirin. β-sitosterol, the polymer gutta-percha, squalene and eicosanoic acid were also isolated. The chemical structures of these constituents were established by IR, 1H and 13C NMR spectral data. Crude extracts and the triterpenes were tested against Entamoeba histolytica, Giardia lamblia and Trichomonas vaginalis and no activity was observed under the in vitro assay conditions. The hexane, chloroform, ethyl acetate and ethanol crude extracts, and the constituent 3,4-seco-friedelan-3-oic acid and 28,29-dihydroxyfriedelan-3-one showed in vitro antimicrobial activity against Salmonella typhimurium, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus cereus, Listeria monocytogenes, Streptococcus sanguinis and Candida albicans.

Enteroparasitoses, Anemia e Estado Nutricional em Grávidas Atendidas em Serviço Público de Saúde

Objetivos: estimar a freqüência de enteroparasitoses em gestantes de pré-natal de baixo risco e sua associação com anemia, estado nutricional, escolaridade e saneamento (fossa sanitária) no domicílio. Métodos: a partir de amostra de 316 gestantes que iniciaram o pré-natal de baixo risco do ambulatório de pré-natal do Instituto Materno Infantil de Pernambuco (IMIP) no período de maio de 2000 a julho de 2001, determinaram-se, por meio de um desenho de corte transversal, as freqüências de enteroparasitoses (método Hoffman, em única amostra) e de anemia (Hb <11,0 g/dL), o estado nutricional (índice de massa corporal - IMC - ajustado para idade gestacional) e indicadores sociais (escolaridade e presença de fossa sanitária em domicílio). Resultados: a freqüência de enteroparasitoses entre estas pacientes foi de 37,0%, sendo 31,6% dos casos positivos representados por monoparasitas. As espécies de parasitos mais freqüentes foram Entamoeba histolytica (13,3%) e Ascaris lumbricoides (12,0%). Detectaram-se 55,4% de gestantes anêmicas, 25,0% de desnutridas e 24,1% com sobrepeso ou obesidade. Houve associação estatisticamente significativa entre enteroparasitose e escolaridade. Não se observou, por outro lado, associação entre anemia, estado nutricional materno, presença de fossa sanitária em domicílio e enteroparasitoses. Conclusões: constataram-se freqüência elevada de enteroparasitoses e anemia, sem, entretanto, haver associação entre estas ocorrências. A escolaridade esteve estatisticamente relacionada com a presença de parasitos intestinais.

Caracterización de la actividad antiamibiana del gosipol

Tesis (Doctorado en Ciencias Especialidad en Microbiología) UANL

In Vitro and In Vivo Antiplasmodial Activities of Risedronate and Its Interference with Protein Prenylation in Plasmodium falciparum

The increasing resistance of malarial parasites to almost all available drugs calls for the identification of new compounds and the detection of novel targets. Here, we establish the antimalarial activities of risedronate, one of the most potent bisphosphonates clinically used to treat bone resorption diseases, against blood stages of Plasmodium falciparum (50% inhibitory concentration [IC(50)] of 20.3 +/- 1.0 mu M). We also suggest a mechanism of action for risedronate against the intraerythrocytic stage of P. falciparum and show that protein prenylation seems to be modulated directly by this drug. Risedronate inhibits the transfer of the farnesyl pyrophosphate group to parasite proteins, an effect not observed for the transfer of geranylgeranyl pyrophosphate. Our in vivo experiments further demonstrate that risedronate leads to an 88.9% inhibition of the rodent parasite Plasmodium berghei in mice on the seventh day of treatment; however, risedronate treatment did not result in a general increase of survival rates.

Aleitamento e infecções por parasitas intestinais em gestantes adolescentes e seus filhos

The exclusive breastfeeding in the first six months of life practice is essential for the health and development of children. Studies show there is in human milk protective antibodies against intestinal parasites and a relationship between the absence of breastfeeding or weaning and parasitic infections. This work was a prospective cohort study involving 34 pregnant adolescents attended school in Maternity Januario Cicco / RN and their children, to assess the influence of breastfeeding on intestinal parasites in them. Thus, the parasitic infection was investigated by examination of feces parasitological and environmental factors by use of questionnaires. The average age of the mothers studied was 16.2 years. Of these 76.4% were infected and the most prevalent species of parasites were Entamoeba histolytica/dispar (76.9%), Giardia lamblia (19.2%) and Ascaris lumbricoides (11.5%). The infected children had an average age of 5.1 months and the frequency of parasites was 61.7%. The infection was earlier detecctada with 45 days of life. The most common parasites were Giardia lamblia (40%), Entamoeba histolytica/dispar (35%), Ascaris lumbricoides (5%) and Ancylostoma (5%). The average length of exclusive breastfeeding was 2.2 months. There was an association between increased duration of exclusive breastfeeding and increased time to detection of parasites in the feces of children. There was no statistically significant correlation between the socio-economic conditions and cultural and breastfeeding. The data reinforce the importance of breastfeeding to protect the children in its first year of life

Ocorrência de enteroparasitos em centros de educação infantil no Município de Patos de Minas, MG, Brasil

Considering the high prevalence of the parasitic diseases in Brazil and its close relationship with the socio-economic and cultural levels of the population, aimed to establish the occurrence of enteroparasites in children of six Centers of Education Child (CEIs) of the municipality of Patos of Minas (MG). Were collected three samples of faeces of 161 children from zero to six years of age. Were used the Lutz and Baermann-Moraes' methods, to the detection of cysts of protozoa, eggs and larvae of helminths. Coproparasitological analyses showed positivity of 73%. The most frequent helminths were: Ascaris lumbricoides (50%), hookworm (22%) and Enterobius vermicularis (0.6%). Among the protozoa highlighted were: Entamoeba histolytica/E. dispar (22%) and Giardia lamblia (32%). The high occurrence of enteroparasites evidenced the need for an effective health policy in the CEIs of the municipality of Patos of Minas to the fight against intestinal parasites.

Parasitoses em lactentes de 0-12 meses atendidos na Faculdade de Medicina de Botucatu - São Paulo

Infestation by protozoa and/or helminths is considered to be extremely rare in infants. We therefore reviewed the records of all infants up to 12 mth of age attending the Paediatric Department from January 1973 to June 1977, in order to assess the frequency of stool examinations for parasites within this age group, the percentage of positive results in these examinations, and some personal characteristics which propitiated parasitic infestation. Some characteristics of the investigated infants were compared to those of a representative sample of the total number of infants attending during the same period. Stool examinations were realized in 11.8% of the 1,162 attending patients and of those, 15.3% were positive. The proportion of positive cases became 24.0% when the infants in whom there was a history of parasites being passed were included. Ascaris lumbricoides and Giardia lamblia were the parasites most frequently found, respectively in 42.8% and 37.1% of the positive cases. The only detected difference between the control group and the group of infants who had parasitologic examinations was the presence of diarrhea, significantly more frequent in the investigated group. The proportion of positive cases was significantly greater in girls, in infants older than 6 mth and when diarrhea was present for up to 15 days, than in boys, in infants up to 6 mth of age and in chronic diarrhea, respectively. Our results show that parasites were investigated in only a small percentage of infants under 1 yr of age, but that in this age group already, parasitic infestation does occur with a certain frequency. As investigated cases were selected, mainly due to the presence of diarrhea, the real prevalence of parasitism in infants under 1 yr of age was impossible to evaluate.

Amoebicidal compounds from medicinal plants.

A series of the natural constituents with amoebicidal activity isolated from several medicinal plants is shown. A list of the medicinal plants potentially active as amoebicide and/or against dysentery also is demonstrated. The present data grouping of the natural compounds and medicinal plants can be an important source of information for the selection of research plant material by the investigators interested in the discovery of new biologically active compounds as amoebicide.