Prevalence and genetic characterization of Dientamoeba fragilis in asymptomatic children attending daycare centers

In order to provide additional data on the prevalence and genetic diversity of Dientamoeba fragilis in human populations, we conducted a study in children from low-income communities in Sao Paulo State, Brazil. Fecal samples from daycare center attendees up to 6 years old (n=156) and staff members (n=18) were submitted to PCR and sequencing of D. fragilis as well as to microscopic examination for the presence of other intestinal parasites. All children assessed were asymptomatic and 10.3% (16/156) were positive for D. fragilis. No worker was found to be positive. An association between Dientamoeba and coinfection with other intestinal parasites was observed. Concerning the genetic diversity, 14 and only two isolates were genotype 1 and genotype 2, respectively. Our findings outline interesting aspects: (1) asymptomatic children as carriers of Dientamoeba in communities in which environmental conditions ensure parasite transmission and, (2) association between Dientamoeba infection in young children and coinfection with other enteric parasites, reinforcing its transmission via the fecal–oral route

Molecular Characterization Of Intestinal Protozoa In Two Poor Communities In The State Of São Paulo, Brazil.

Several species of protozoa cause acute or chronic gastroenteritis in humans, worldwide. The burden of disease is particularly high among children living in developing areas of the world, where transmission is favored by lower hygienic standards and scarce availability of safe water. However, asymptomatic infection and polyparasitism are also commonly observed in poor settings. Here, we investigated the prevalence of intestinal protozoa in two small fishing villages, Porto Said (PS) and Santa Maria da Serra (SM), situated along the river Tietê in the State of São Paolo, Brazil. The villages lack basic public infrastructure and services, such as roads, public water supply, electricity and public health services. Multiple fecal samples were collected from 88 individuals in PS and from 38 individuals in SM, who were asymptomatic at the time of sampling and had no recent history of diarrheal disease. To gain insights into potential transmission routes, 49 dog fecal samples (38 from PS and 11 from SM) and 28 river water samples were also collected. All samples were tested by microscopy and PCR was used to genotype Giardia duodenalis, Blastocystis sp., Dientamoeba fragilis and Cryptosporidium spp. By molecular methods, the most common human parasite was Blastocystis sp. (prevalence, 45% in PS and 71% in SM), followed by D. fragilis (13.6% in PS, and 18.4% in SM) and G. duodenalis (18.2% in PS and 7.9% in SM); Cryptosporidium spp. were not detected. Sequence analysis revealed large genetic variation among Blastocystis samples, with subtypes (STs) 1 and 3 being predominant, and with the notable absence of ST4. Among G. duodenalis samples, assemblages A and B were detected in humans, whereas assemblages A, C and D were found in dogs. Finally, all D. fragilis samples from humans were genotype 1. A single dog was found infected with Cryptosporidium canis. River water samples were negative for the investigated parasites. This study showed a high carriage of intestinal parasites in asymptomatic individuals from two poor Brazilian villages, and highlighted a large genetic variability of Blastocystis spp. and G. duodenalis.810

Prevalência de parasitas intestinais e caracterização genotípica de Giardia duodenalis em creche do município de Pratânia, estado de São Paulo

O presente estudo foi realizado para investigar a prevalência de enteroparasitas em crianças e funcionários da creche do município de Pratânia, Estado de São Paulo, e caracterizar geneticamente os isolados de Giardia duodenalis obtidos dos indivíduos desse grupo empregando gdh e tpi como genes-alvo. Para isso, amostras de fezes de 141 crianças com idade de zero a seis anos e de 18 colaboradores foram colhidas e processadas pelo método de centrífugo-flutuação e pelo kit TF-test®. Nas 141 crianças, parasitas e/ou comensais intestinais foram diagnosticados em 64 (45,4%), das amostras, sendo Giardia (22%) e Cryptosporidium (22%) os parasitas mais freqüentes. Nos funcionários, Giardia foi diagnosticada em apenas uma amostra (5,5%) e o protozoário comensal Entamoeba coli (22,2%) foi o mais prevalente. Empregando técnicas baseadas em PCR, o DNA extraído de 86 (31 amostras positivas e 55 negativas para Giardia no exame microscópico) foi amplificado e os produtos obtidos foram seqüenciados. A análise de 35 seqüências revelou a ocorrência de infecção pelos genótipos A (28,6%) e B (71,4%), subtipos AI, AII, BIII/BSI e BIV, no entanto, associações entre os genótipos e fatores de risco para a infecção não foram observadas. O alinhamento das seqüências de gdh e tpi, revelou a presença de um a 12 SNPs (Single Nucleotide Polymorphisms) em isolados dos genótipos A e B, sendo que em alguns deles, as mutações foram responsáveis por alterações nas seqüências de aminoácidos. A infecção de seis crianças de um mesmo grupo e de uma funcionária por isolados classificados como genótipo B e apresentando seqüências nucleotídicas 100% idênticas entre si sugere a ocorrência de transmissão antroponótica na população estudada. Além disso, a maior prevalência do genótipo B, grupo predominante nas infecções humanas, reforça o fato de que a transmissão...The present study was conducted to investigate the prevalence of intestinal parasites in children and employees of a daycare center of Pratania, São Paulo State, and to evaluate the genotypes Giardia duodenalis isolates obtained from individuals in this group by using gdh and tpi genes as targets. Fecal samples of 141 children aged zero to six years and 18 employees were processed by a flotation method and the TF-Test® kit. Of the 141 children samples, parasites and/or commensal organisms were diagnosed in 64 (45.4%) samples, and Giardia (22%) and Cryptosporidium (22%) were the most frequent parasites. In employees, Giardia was diagnosed in only one sample (5.5%) and the commensal protozoan Entamoeba coli (22.2%) was the most prevalent. DNA extracted from 86 samples (31 samples positive and 55 negative for Giardia by microscopic examination) was amplified by PCR and the products were sequenced. The analysis of 35 sequences revealed the occurrence of infection with genotypes A (28.6%) and B (71.4%) subtypes AI, AII, BIII / BIV and BSI, however, associations between genotypes and risk factors for infection were not observed. The alignment of the sequences revealed SNPs (Single Nucleotide Polymorphisms) in both genotypes A and B, altering the amino acid sequence of some isolates. Among isolates identified as genotype B, six obtained from children of the same group and one from the only one employee infected with Giardia showed gdh and tpi sequences different from reference ones, but 100% identical to each other. This data is suggestive of anthroponotic transmission in the studied population. In addition this finding, the higher prevalence of genotype B, the main group associated to human infections, reinforces the fact that anthroponotic transmission has importance in collective environments that meet children at an age that they still have not practice the basic precepts of hygiene.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

EFFECT OF Bifidobacterium animalis ON MICE INFECTED WITH Strongyloides venezuelensis

The administration of viable Bifidobacterium animaliswas tested to induce resistance against Strongyloides venezuelensis infection in mice. Effects on parasite burden, worm length, egg output, and intestinal mucosal histology were evaluated. The oral administration of B. animalis, strain 04450B, starting 14 days before the inoculation of nematode larvae significantly decreased the worm burden and egg output. In probiotic treated animals, the percent reduction of adult worms in the intestine was of 33% and the reduction of egg production was of 21%, compared with those of the control group. The duodenum villous height and villous/crypt ratio were significantly higher in probiotic-treated mice, indicating that this group could be experiencing less intestinal damage. The present findings revealed that the administration of B. animalis for the amelioration of host response to nematode infections is biologically plausible and could have some potential for impacting public health. Meanwhile, further study is needed to delineate the nature and identity of the factor(s) involved in these beneficial effects