RESUMO
Giardia duodenalis is one of the most important intestinal parasites globally, especially in children, and in Cuba is the leading cause of chronic paediatric diarrhoea in this population. G. duodenalis is composed of eight genetic groups (or assemblages), two of which (A and B) are apparently zoonotic, occurring in both humans and other animals. However, consensus on the most appropriate genotyping scheme for optimal characterization of G. duodenalis isolates is lacking. In this article we present the results of three descriptive observational studies conducted in Havana, Cuba between 2010 and 2013, with the aim of comparing the results from molecular (PCR) approaches targeting different genes in order to assign with confidence 224 isolates of G. duodenalis to the correct assemblages. In each sub-study, following DNA isolation by the phenol/chloroform/isoamyl alcohol extraction method, PCR targeting the triose phosphate isomerase (tpi) gene was used for molecular characterization, as well as one additional PCR-method targeting another gene or pair of genes. DNA amplification was obtained in 87%, 83%, and 80% in the three sub-studies. Although excellent agreement (kappa index = 1) was recorded between results from some pairs of genes, for other combinations only moderate or substantial agreement was achieved. These results highlight the importance of interpretation of genotyping data, especially when single genetic markers are used. From the results of our studies, PCR targeting a combination of the tpi gene and the intergenic spacer region of rDNA may be a useful approach for the molecular characterization of G. duodenalis isolates.
Assuntos
Técnicas de Genotipagem/normas , Giardia lamblia/classificação , Giardíase/parasitologia , Reação em Cadeia da Polimerase/métodos , Animais , Criança , Pré-Escolar , Cuba , DNA de Protozoário/química , DNA de Protozoário/isolamento & purificação , DNA Espaçador Ribossômico/química , Fezes/parasitologia , Giardia lamblia/genética , Giardia lamblia/isolamento & purificação , Glutamato Desidrogenase/genética , Humanos , Polimorfismo de Fragmento de Restrição , Triose-Fosfato Isomerase/genéticaRESUMO
Giardia duodenalis is one of the most frequent intestinal parasitic infections in children worldwide. To date, eight main assemblages of G. duodenalis have been described, but only A and B genetic groups are known to infect humans. A cross-sectional study was conducted to determine the prevalence and risk factors of Giardia duodenalis infection in 417 preschool children from the Fomento municipality in the central region of Cuba between January and June 2013. The overall prevalence of Giardia infection was 10.79 %. Assemblage identification was carried out by the amplification of a fragment of the triose phosphate isomerase (tpi) gene. DNA from 36 of 45 (80 %) samples was successfully amplified by PCR-tpi. Assemblage B and mixed assemblages A + B represented 52.78 and 36.11 % respectively, of genotyped samples. Assemblage A accounts for only 11.11 %. Children who were cared for at home were associated with diarrhea caused by assemblage B. No associations were found between other clinical variables with infecting assemblage of Giardia. Univariate analysis identified the use of unsafe water resources (OR 2.9; CI 1.2-6.8) and-even more interestingly-keeping dogs indoor (OR 2.5; CI 1.2-5.4) were significant risk factors associated with Giardia infection among children. Multivariate analysis using introduction test logistic regression ratified the association of these two risk factors: kept dogs indoor (OR 2.8, CI 1.1-5.3), and untreated water (OR 1.4, CI 1.4-4.9) with Giardia infection. This information may be useful for an effective prevention and control programme of giardiasis in this population.
RESUMO
Giardia duodenalis is a worldwide protozoan parasite that infects humans and other mammals including dogs. Due to the risk of zoonotic transmission between dogs and humans, we aimed in this study to determine the prevalence of the intestinal parasites and the distribution of assemblages of G. duodenalis among dogs analysed. A descriptive cross-sectional study was carried out in La Habana from June 2014 to March 2015 in the Zoonosis Unit of La Lisa municipality. A total of 98 dogs were analysed by three different techniques (microscopy with faecal concentration, Enzyme-Linked ImmunoSorbent Assay, and Polymerase Chain Reaction) in order to detect Giardia in stool samples. Out of 98 dogs studied, 43 (43.9%) were infected with intestinal parasites. The zoonotic parasites Ancylostoma caninum (21.4%), Trichuris vulpis (16.3%) and the protozoan Giardia duodenalis (11.2%) were the most prevalent parasites. In regards to the G. duodenalis, seven dogs were positive by microscopy after faecal concentration, nine by NOVITEC® Giardia Microplate Assay, and ten and eleven samples were amplified by the ß-giardin and SSU-RNA PCRs, respectively. After PCR sequence analysis of both genes only zoonotic assemblages (A and B) were detected. The SSU-RNA sequence results revealed a distribution of 8 assemblage A and 4 assemblage B, whereas only assemblage A were identified by the ß-giardin analysis. Among subassemblage classification by ß-giardin phylogenetic tree, four isolates showed an AI pattern and one isolate displayed an AII distribution. Mixed infections were detected in three isolates. These findings highlight the risk of zoonotic transmission of Giardia duodenalis between dogs and humans.
RESUMO
Giardiasis is considered the most common intestinal parasitic disease in humans worldwide. In Cuba, this infection has particularly a strong clinical impact on the child population. Giardia duodenalis is a highly diverse protozoan, which comprises a complex of eight morphologically identical genetic assemblages, further divided into sub-assemblages. The present study used triose phosphate isomerase (tpi) and small-subunit ribosomal RNA (SSU rRNA) genes as genetic markers for the identification of G. duodenalis assemblages and sub-assemblages in correlation with clinical and epidemiological data in children attended at the Paediatric Hospital "William Soler" and at Pedro Kouri Institute, between 2015 and 2016. A prevalence of 8% of G. duodenalis infection was recorded in stool samples after concentration techniques from 68 children out of 847 analysed. A 100% detection of Giardia DNA was achieved by a SSU-rRNA PCR, whereas DNA from 63 of 68 (92.6%) was successfully amplified by tpi-PCR. By this assemblage-specific tpi-PCR 32 (50.8%) assemblage B, 17 (27.0%) assemblage A and 14 (22.2%) mixed infection (A + B) were identified. Assemblage B was significantly (P < 0.02) more frequently found in children with diarrhoea. Sequence analysis of the tpi gene of Giardia isolates from symptomatic children showed that assemblage A belonged to the sub-assemblage AII, and 4 sub assemblages BIV and 1 sub assemblage BIII were also recorded. Only 2 discordant genotyping results were observed by phylogenetic comparison of SSU-rRNA and tpi sequences. Further studies with novel molecular tools for a better discrimination at the sub-assemblage level are needed to identify the dynamics of spread of giardiasis and to verify possible correlations between Giardia genetic diversity and clinical manifestation.
RESUMO
Soil-transmitted helminth (STH) infections and zinc deficiency are often found in low- and middle-income countries and are both known to affect child growth. However, studies combining data on zinc and STH are lacking. In two studies in schoolchildren in Cuba and Cambodia, we collected data on height, STH infection and zinc concentration in either plasma (Cambodia) or hair (Cuba). We analyzed whether STH and/or zinc were associated with height for age z-scores and whether STH and zinc were associated. In Cuba, STH prevalence was 8.4%; these were mainly Ascaris lumbricoides and Trichuris trichiura infections. In Cambodia, STH prevalence was 16.8%, mostly caused by hookworm. In Cuban children, STH infection had a strong association with height for age (aB-0.438, p = 0.001), while hair zinc was significantly associated with height for age only in STH uninfected children. In Cambodian children, plasma zinc was associated with height for age (aB-0.033, p = 0.029), but STH infection was not. Only in Cambodia, STH infection showed an association with zinc concentration (aB-0.233, p = 0.051). Factors influencing child growth differ between populations and may depend on prevalences of STH species and zinc deficiency. Further research is needed to elucidate these relationships and their underlying mechanisms.
Assuntos
Estatura , Helmintíase/sangue , Helmintíase/epidemiologia , Solo/parasitologia , Zinco/sangue , Animais , Ascaris lumbricoides , Camboja/epidemiologia , Criança , Desenvolvimento Infantil , Estudos Transversais , Cuba/epidemiologia , Países em Desenvolvimento , Feminino , Cabelo/química , Helmintíase/transmissão , Humanos , Modelos Lineares , Masculino , Prevalência , Trichuris , Zinco/administração & dosagem , Zinco/deficiênciaRESUMO
Giardia duodenalis is one of the most frequent intestinal parasitic infections in children worldwide. To date, eight main assemblages of G. duodenalis have been described, but only A and B genetic groups are known to infect humans. In Cuba, this parasite has most clinical impact on children. The aim of this investigation was genetic characterization of G. duodenalis isolated from children with giardiasis diagnosed at the Paediatric Hospital "William Soler" between 2010 and 2011, and to compare the genetic results with clinical and epidemiological data. A total of 103 stool samples from 452 children were positive for G. duodenalis and co-infections with other parasites were noted in 5 cases. Assemblage identification was carried out by the amplification of a fragment of the triosephosphate isomerase (tpi) gene. Sub-assemblages of assemblage A (AI and AII) were identified by a nested PCR using the intergenic spacer (IGS) region of ribosomal deoxyribonucleic acid gene as a target. DNA from 90 of 103 (87.4%) samples was successfully amplified by PCR-tpi. The prevalence of assemblages A and B was 40% and 42%, respectively. Infections with both assemblages were reported in 16 cases. No associations between epidemiological information and assemblage was detected, but assemblage B was significantly (P<0.01) more frequently found in children with diarrhea, flatulence or abdominal pain than assemblage A. Sub-assemblage AII accounted for the majority of cases (86.5%).
Assuntos
Giardia lamblia/genética , Giardia lamblia/isolamento & purificação , Giardíase/epidemiologia , Giardíase/parasitologia , Criança , Pré-Escolar , Coinfecção/epidemiologia , Cuba/epidemiologia , Fezes/parasitologia , Feminino , Genes de Protozoários , Giardia lamblia/classificação , Humanos , Masculino , Análise de Sequência de DNA , Triose-Fosfato Isomerase/genéticaRESUMO
Although the population of Hispaniola, Cuba's most similar neighbour in the Caribbean, continues to be threatened by parasitic diseases (including malaria), many tropical parasitic infections in Cuba have been eliminated or controlled. However, some parasitic infections remain important in the Cuban population, and the occurrence of vectors and the high possibility of introduction of parasites mean that Cuban diagnosticians must remain alert. Some key aspects of human parasitology in Cuba are reviewed here, including historical information, comparative data from Hispaniola and Jamaica, and how Cuba strives to maintain and improve its control against parasitic infections. Data from recent key novel parasitology research conducted in Cuba are also described.
Assuntos
Política de Saúde , Enteropatias Parasitárias/epidemiologia , Programas Nacionais de Saúde , Animais , Comparação Transcultural , Cuba/epidemiologia , Países em Desenvolvimento , Diagnóstico Diferencial , Humanos , Enteropatias Parasitárias/diagnóstico , Enteropatias Parasitárias/tratamento farmacológico , SocialismoRESUMO
OBJECTIVES: To determine the prevalence of intestinal parasite infections and their risk factors in children in urban and rural settings in two Cuban municipalities. METHODS: A total of 1320 Cuban schoolchildren aged 4-14 were tested by stool examination for intestinal parasite infections and evaluated by parental questionnaire for a number of common environmental, sanitary, socioeconomic and behavioural risk factors. Multivariate regression was applied to examine the relationship between the respective parasite infections and the risk factors. RESULTS: Prevalences of intestinal parasite infections were 58% in Fomento and 45% in San Juan y Martínez; for helminth infections, these were 18% and 24% and for protozoa infections, 50% and 29%, respectively. Helminth infections were associated with high parental education (maternal: OR 0.68, CI 0.50-0.93; paternal: OR 0.71, CI 0.52-0.96), absence of toilet (OR 1.57, CI 1.12-2.19), consumption of water from a well or river (OR 0.56, CI 0.41-0.77) and eating unpeeled/unwashed fruit (OR 1.37, CI 1.01-1.87); protozoa infections were only associated with high maternal education (OR 0.72, CI 0.57-0.91). CONCLUSIONS: Paediatric intestinal parasite infections are still prevalent in certain areas in Cuba and associated with a number of common environmental, socioeconomic and sanitary risk factors.
Assuntos
Enteropatias Parasitárias/epidemiologia , Adolescente , Criança , Pré-Escolar , Estudos Transversais , Cuba/epidemiologia , Escolaridade , Feminino , Humanos , Enteropatias Parasitárias/etiologia , Enteropatias Parasitárias/transmissão , Masculino , Prevalência , Fatores de Risco , Saúde da População Rural/estatística & dados numéricos , Saneamento/estatística & dados numéricos , Fatores Socioeconômicos , Saúde da População Urbana/estatística & dados numéricosRESUMO
275 adolescents aged 10-19 receiving attention at the infantojuvenile department of pregnancy interruption and of sexually transmitted infections of "Ramón Gonzalez Coro" and "Eusebio Hernández" Gynecoobstetric Teaching Hospitals, both in Havana City, were studied from August, 1999, to September, 2000, aimed at knowing the behavior of the infection caused by Trichomonas vaginalis in this population group. The results yielded 18.1 % of positivity to parasitosis. It was proved by the calculation of relative risk that the adolescents that had an active or past infection due to Trichomonas vaginalis have more possibilities of getting an infection produced by human Papilomavirus than those without this condition. It was demonstrated that 78 (28.3 %) of the total of patients examined had some type of lesion in the uterine cervix. Some risk factors as non using condon, having multiple sexual partners and the early beginning of sexual relations, which may facilitate the adquisition of this parasitosis in a population so vulnerable as the adolescents, were made known.
Assuntos
Vaginite por Trichomonas/diagnóstico , Adolescente , Adulto , Criança , Feminino , HumanosRESUMO
The results of the National Toxoplasmosis Reference Laboratory during 1999 and 2000 were processed and the evaluations on this topic were simultaneously analyzed among 51 professionals that attended the First National Workshop held at "Pedro Kouri" Institute of Tropical Medicine at that time. Of 3 672 orders, 3 171 (86.3%) did not have the reason for serology, 36.1% had no place of origin and only 301 cases (8.1%) were indicated the matched serum. On analyzing the main causes leading to make this test, 13.5% corresponded to repeated abortions, which is not accepted at present as a clinical manifestation in the course of the disease. Coincidentally, when some professionals were evaluated through a survey about the basic aspects related to this parasitosis, only 31.3% were able to obtain satisfactory marks. Most of the mistakes were observed in the interpretation of the laboratory results.
Assuntos
Toxoplasmose/diagnóstico , Cuba , Educação Médica , Humanos , Parasitologia/educaçãoRESUMO
One-hundred and seventy two couples which went for the first time to the Infertility Service of the National Institute of Endocrinology from June 1999 to June 2000, were studied to find out the frequency of Trichomonas vaginalis infection in this group, and determine its interaction with a number of clinical and risk variables. The results yielded that 10.5% were positive to the parasite, the prevailing symptom was leukorrea in women and 96.6% of men showed no symptoms. It was highly significant the fact of having a previous pathological history that might be related to infertility and current T. vaginalis infection. This protozoon was frequently associated with Candida sp. and causative agents of bacterial vaginosis in women and Haemophilus influenzae in men. This parasite seems to play an important role as a likely causative agent to be considered in fertility problems.
Assuntos
Infertilidade/parasitologia , Tricomoníase/complicações , Tricomoníase/epidemiologia , Trichomonas vaginalis , Adulto , Animais , Feminino , Humanos , MasculinoRESUMO
A total of 132 women who received attention at the Outpatient Department of Dermatology of the "Pedro Kouri" Institute of Tropical Medicine from January to July, 1998, were studied. 64 of them were HIV carriers and 68 were sound controls. On determining the infection frequency by Trichomonas vaginalis, it was found that 15.6 and 16.1, respectively, were parasitized by this protozoa. The diagnostic techniques used were simple direct examination of the vaginal exudate and culture in vitro. The latter proved to be more sensitive on yielding 100 of sensitivity. It was determined that 48 hours was the optimum time for reading each specimen.