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BACKGROUND: Surveys for intestinal parasitic infections (IPIs) often involve samples from three sampling dates analysed by various microscopy techniques. However, analysis of three samples per individual is a huge burden on time and resources. We compared the value from analysing three or fewer samples. METHODS: In this cross-sectional study, three faecal samples were collected every other day from 332 children from two locations in Guantanamo province, Cuba. Samples were analysed by wet mount with Lugol stain, Willis flotation method and Kato-Katz thick smear. RESULTS: Most parasites were detected by wet mount, although helminth eggs not found by wet smear were detected by the Willis flotation method (in particular) and Kato-Katz smear. Blastocystis spp. was the most commonly detected parasite (about 65%), then Giardia duodenalis and then Entamoeba spp. Although analysis of two stool samples significantly increased occurrence data for Blastocystis, this was not so for the other parasites. For none of the protozoan parasites were results from analysing three samples significantly higher than results from analysing just two samples. CONCLUSIONS: Analysing two faecal samples by wet mount and the Willis flotation method provides useful data for estimating the prevalence of IPIs in relatively high prevalence settings. Analysing further samples provides limited additional information and adds an extra burden in terms of time and resources.
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Giardia lamblia , Helmintos , Parasitosis Intestinales , Parásitos , Animales , Niño , Humanos , Estudios Transversales , Parasitosis Intestinales/epidemiología , Parasitosis Intestinales/parasitología , Heces/parasitología , PrevalenciaRESUMEN
The aetiology of diarrhoea in a patient in Cuba with HIV was investigated. Although molecular diagnostics are still not used in many under-resourced settings, here traditional methods were supported by use of PCR. This approach enabled detection of a dual infection (Cystoisospora belli and Enterocytozoon bieneusi), the latter of which was not identified by microscopy with Didier's trichromic staining.
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Coccidiosis/diagnóstico , Diarrea/diagnóstico , Enterocytozoon/aislamiento & purificación , Microsporidiosis/diagnóstico , Sarcocystidae/aislamiento & purificación , Adulto , Antiinfecciosos/uso terapéutico , Coccidiosis/tratamiento farmacológico , Cuba , Diarrea/microbiología , Diarrea/parasitología , Enterocytozoon/genética , Infecciones por VIH/complicaciones , Infecciones por VIH/tratamiento farmacológico , Humanos , Huésped Inmunocomprometido , Masculino , Microsporidiosis/tratamiento farmacológico , Técnicas de Diagnóstico Molecular , Reacción en Cadena de la Polimerasa , Sarcocystidae/genética , Sarcoma de Kaposi/complicaciones , Sarcoma de Kaposi/tratamiento farmacológico , Combinación Trimetoprim y Sulfametoxazol/uso terapéuticoRESUMEN
Taeniosis is a neglected disease, particularly in developing countries, and is caused by infection with the adult tapeworm of either Taenia solium, Taenia saginata, and Taenia asiatica. Of these, T. solium is of primary concern due to the potential for cysticercosis should T. solium eggs be ingested. In Cuba, all cases of taeniosis are assumed to be caused by T. saginata, although some cases of cysticercosis have been documented. It is therefore important to gain further insights regarding the species causing taeniosis in Cuba, especially as diagnostic records indicate an increasing incidence, with the highest number of cases reported in 2020. In this study, we analysed 37 Taenia-positive faecal samples (or proglottids isolated from faecal samples) from the period 2001 until 2020 from all regions of the country. Genomic DNA was extracted from the samples, which had been stored in 10% formalin, using the QIAamp Tissue Kit. Species identification was carried out by duplex real-time PCR targeting the mitochondrial DNA. All cases were found to be T. saginata, and sequence analysis of three isolates confirmed the identification of this species. Our data do not provide any evidence that T. solium currently occurs in Cuba. However, given the relatively low number of samples analysed here, that the parasite may be imported with visitors or travellers who have been in endemic countries, and that taeniosis has relatively mild symptoms and thus infected patients may not seek medical attention, we recommend species determination for all taeniosis cases reported in Cuba.
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To investigate the presence of Cyclospora cayetanensis, Toxoplasma gondii and Echinococcus spp. in fresh produce sold in Italy, 324 locally produced 'ready-to-eat' (RTE) mixed-salad packages belonging to three brands and 324 berries packages (blueberries and blackberries imported from Peru and Mexico, respectively, and raspberries grown in Italy) were purchased at retail. Nine individual packages from each of the six types of fresh produce were collected monthly for one year, and with the same produce pooled, this resulted in a total of 72 pools for the whole year. Using microscopy (FLOTAC), a Cyclospora-like oocyst was detected in a blueberry sample and a taeniid egg was detected in a RTE-salad sample. Molecular tools confirmed these to be C. cayetanensis and Echinococcus multilocularis, respectively. Toxoplasma gondii was not detected in any of the samples. This study shows for the first time in Europe that imported berries on the Italian market may be contaminated with C. cayetanensis and RTE salads grown in Italy with E. multilocularis. The results indicate a new epidemiological scenario and highlight that current management of fresh produce, locally produced or imported, does not ensure products are free from parasite contamination.
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Cyclospora/crecimiento & desarrollo , Echinococcus multilocularis/crecimiento & desarrollo , Comida Rápida/parasitología , Contaminación de Alimentos/análisis , Frutas/parasitología , Animales , Arándanos Azules (Planta)/parasitología , Cyclospora/genética , Cyclospora/aislamiento & purificación , Echinococcus multilocularis/genética , Echinococcus multilocularis/aislamiento & purificación , Italia , México , Oocistos/genética , Oocistos/aislamiento & purificación , Rubus/parasitología , Toxoplasma/genética , Toxoplasma/crecimiento & desarrollo , Toxoplasma/aislamiento & purificaciónRESUMEN
Microscopy is the gold standard for diagnosis of intestinal parasitic diseases in many countries, including Cuba, although molecular approaches often have higher sensitivity as well as other advantages. Fecal samples from 133 patients were analyzed by light microscopy and also real-time multiplex qPCR targeting Giardia duodenalis, Cryptosporidium spp., and Entamoeba histolytica, and, separately, Dientamoeba fragilis. Microscopy revealed G. duodenalis occurred most commonly (17 patients), followed by Blastocystis spp. (12 patients). In a few patients, Entamoeba histolytica/E. dispar, Cryptosporidium spp., and Cyclospora cayetanensis were identified. Molecular analysis identified 4 more G. duodenalis infections and 2 more Cryptosporidium spp. infections; concordance between microscopy and PCR showed almost perfect agreement for G. duodenalis (κ = 0.88) and substantial agreement for Cryptosporidium (κ = 0.74). PCR indicated that E. dispar, rather than E. histolytica, had been identified by microscopy. Additionally, 16 D. fragilis infections were detected using molecular methods. Although both microscopy and molecular techniques have a place in parasitology diagnostics, for parasites such as D. fragilis, where microscopy can underestimate occurrence, molecular techniques may be preferable, and also essential for distinguishing between morphologically similar microorganisms such as E. histolytica and E. dispar. Although in resource-constrained countries such as Cuba, microscopy is extremely important as a diagnostic tool for intestinal parasites, inclusion of molecular techniques could be invaluable for selected protozoa.
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Criptosporidiosis/diagnóstico , Dientamebiasis/diagnóstico , Entamebiasis/diagnóstico , Giardiasis/diagnóstico , Parasitosis Intestinales/diagnóstico , Adolescente , Adulto , Anciano , Niño , Preescolar , Estudios Transversales , Criptosporidiosis/parasitología , Cryptosporidium/aislamiento & purificación , Cuba/epidemiología , Dientamoeba/aislamiento & purificación , Dientamebiasis/parasitología , Entamoeba histolytica/aislamiento & purificación , Entamebiasis/parasitología , Heces/parasitología , Femenino , Giardia lamblia/aislamiento & purificación , Giardiasis/parasitología , Humanos , Parasitosis Intestinales/epidemiología , Parasitosis Intestinales/parasitología , Masculino , Microscopía/métodos , Persona de Mediana Edad , Técnicas de Diagnóstico Molecular , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Adulto JovenRESUMEN
ABSTRACT: Berries are potential vehicles for the transmission of parasites and have been implicated in illness outbreaks in various countries around the world, particularly in the United States. Although data on contamination of fresh produce with foodborne parasites have been obtained from various global regions, data from Colombia are lacking even though South American countries are major producers of fresh produce, which is both consumed nationally and exported. In this study, we used a previously published multiplex quantitative PCR approach to investigate contamination of strawberries purchased in either supermarkets or local markets in 20 localities. Strawberries were washed in a detergent solution after purchase, and the eluate was concentrated and sent to Norway for molecular analysis. Of the 120 strawberry samples analyzed, wash eluate from 6 samples (5%) tested positive for Toxoplasma gondii DNA, and 1 sample (0.83%) was positive for Cyclospora cayetanensis DNA. These results indicate that strawberries for sale in Bogotá, Colombia, may be contaminated with T. gondii and C. cayetanensis and, therefore, could act as transmission vehicles for these parasites. These data also indicate that cat and human fecal contamination of the strawberries has occurred at some point in their production, transportation, or storage. These findings highlight the need for a hazard analysis critical control point investigation of the strawberry production chain and implementation of measures to reduce the risk of strawberry contamination, thereby minimizing the risk of transmission of parasitic infection via these fruits, which are usually consumed raw.
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Cyclospora , Fragaria , Parásitos , Animales , Gatos , Colombia , NoruegaRESUMEN
In Cuba, there are few studies on cyclosporiasis. Here, we report results from 1247 stool samples from symptomatic patients that were examined by microscopy methods and positive cases confirmed by nested PCR targeting the 18S rRNA gene, followed by sequencing. Seven positive samples, all diagnosed during May-June, were confirmed by the molecular method, indicating an occurrence in this patient cohort of 0.56%.
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Cyclospora/aislamiento & purificación , Ciclosporiasis/diagnóstico , Adulto , Niño , Preescolar , Cuba/epidemiología , Cyclospora/clasificación , Cyclospora/citología , Cyclospora/genética , Ciclosporiasis/epidemiología , Ciclosporiasis/parasitología , ADN Protozoario/genética , Heces/parasitología , Femenino , Humanos , Masculino , Microscopía , Persona de Mediana Edad , Filogenia , Reacción en Cadena de la Polimerasa , Prevalencia , ARN Ribosómico 18S/genética , Estaciones del Año , Análisis de Secuencia de ADN , Adulto JovenRESUMEN
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.
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Técnicas de Genotipaje/normas , Giardia lamblia/clasificación , Giardiasis/parasitología , Reacción en Cadena de la Polimerasa/métodos , Animales , Niño , Preescolar , Cuba , ADN Protozoario/química , ADN Protozoario/aislamiento & purificación , ADN Espaciador Ribosómico/química , Heces/parasitología , Giardia lamblia/genética , Giardia lamblia/aislamiento & purificación , Glutamato Deshidrogenasa/genética , Humanos , Polimorfismo de Longitud del Fragmento de Restricción , Triosa-Fosfato Isomerasa/genéticaRESUMEN
BACKGROUND: The distribution of Taenia saginata in the Americas is unclear. Establishing the distribution, economic burden, and potentials for control of bovine cysticercosis is increasingly important due to the growing demand for beef. This paper aims to take the first step and reviews the recent distribution of T. saginata taeniosis and bovine cysticercosis on a national level within the Americas. METHODS: We undertook a systematic review of published and grey literature for information on the occurrence, prevalence, and geographical distribution of bovine cysticercosis and human taeniosis in the 54 countries and territories of the Americas between January 1st, 1990 and December 31st, 2017. Data on bovine cysticercosis from OIE reports from 1994 to 2005 were also included. RESULTS: We identified 66 papers from the Americas with data on the occurrence of taeniosis or bovine cysticercosis and an additional 19 OIE country reports on bovine cysticercosis. Taeniosis was reported from 13 countries, with nine of these countries reporting specifically T. saginata taeniosis, and four countries reporting non-species specific taeniosis. The reported prevalence of taeniosis ranged between 0.04-8.8%. Bovine cysticercosis was reported from 19 countries, nine identified through the literature search, and an additional 10 identified through the OIE country reports for notifiable diseases. The reported prevalence of bovine cysticercosis ranged between 0.1-19%. Disease occurrence was restricted to 21 countries within the Americas, the majority from the mainland, with the only island nations reporting either bovine cysticercosis or taeniosis being Cuba, Haiti, and the US Virgin Islands. CONCLUSIONS: Taenia saginata is widely distributed across 21 of the 54 countries in the Americas, but insufficient epidemiological data are available to estimate the subnational spatial distribution, prevalence, incidence and intensity of infections. This needs to be addressed through active surveillance and disease detection programmes. Such programmes would improve the data quantity and quality, and may enable estimation of the economic burden due to bovine cysticercosis in the region in turn determining the requirement for and cost-effectiveness of control measures.
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Cisticercosis/epidemiología , Taenia saginata/fisiología , Teniasis/epidemiología , Américas/epidemiología , Animales , Cisticercosis/parasitología , Femenino , Humanos , Incidencia , Masculino , Prevalencia , Teniasis/parasitologíaRESUMEN
Although giardiasis has been a threat to mankind for thousands of years, this protozoan infection was, until recently, relatively neglected. Giardia duodenalis is recognised as a major cause of parasite-induced diarrhoea in humans and animals, and is currently an important public health problem, placing a heavy burden on both diagnostic and treatment services at health care institutions, mostly in developing countries, but also in highly developed countries. Steady progress in recent years, using a combination of molecular, immunological, and clinical approaches, has substantially increased our understanding of Giardia and important aspects of the clinical manifestations that it causes. The purpose of this review is to provide an overview of the extent of Giardia infection, the implications of water and food in Giardia transmission, new aspects regarding clinical diagnosis and environmental detection, treatment, and some approaches towards prevention and control. A number of future research priorities are also presented.
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Giardia lamblia , Giardiasis/diagnóstico , Giardiasis/terapia , Enfermedades Desatendidas/diagnóstico , Enfermedades Desatendidas/terapia , Animales , Enfermedades Transmitidas por los Alimentos/parasitología , Giardia lamblia/aislamiento & purificación , Humanos , Enfermedades Desatendidas/parasitología , Agua/parasitologíaRESUMEN
Fascioliasis, an emerging parasitic infection, impacts significantly on both veterinary and human health worldwide. Endemic foci are not limited only to areas of extensive livestock farming, but owing to the parasite's abilities to colonise new intermediate hosts and adapt to new environments, also occur in other places, including Cuba. In Cuba, despite a high prevalence of fascioliasis in livestock, and the widespread occurrence of two potential intermediate hosts, human infection has decreased steadily over the past 10 years. In other parts of the world, human fascioliasis is apparently becoming more frequent. Problems in counteracting the spread of fascioliasis, and approaches used in Cuba to limit zoonotic transmission are discussed, with emphasis on diagnostic and treatment problems, malacological initiatives, and the importance of an integrated control programme. Such programmes may be of benefit in other countries where the prevalence of human fascioliasis is increasing, and lessons may perhaps be learned from the Cuban approach.
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Enfermedades Transmisibles Emergentes/epidemiología , Enfermedades Transmisibles Emergentes/prevención & control , Fascioliasis/epidemiología , Animales , Cuba/epidemiología , Humanos , PrevalenciaRESUMEN
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.