Detection of Human Paragonimiasis by ELISA Using Recombinant Paragonimus westermani Cysteine Protease 7.

Paragonimiasis is an important but neglected foodborne trematodiasis caused by Paragonimus mexicanus in Costa Rica. Immunological techniques for diagnosing this parasitosis in humans do not exist in Central America. The objective of the present study was to use recombinant Paragonimus westermani cysteine protease 7 to standardize an ELISA for the detection of antibodies against Paragonimus spp. Human sera positive for P. westermani, P. mexicanus, or Paragonimus spp., human sera infected with other helminths, as well as sera of healthy humans without parasitic infections, were analyzed. The sensitivity of the ELISA was 92.9%, and the specificity was 91.9%. This report is the first to describe the development of an ELISA for the diagnosis of Paragonimus spp. in Costa Rica and Central America. Using this ELISA in the health system of Costa Rica is recommended to detect infections.

Pulmonary, liver and cerebral paragonimiasis: An unusual clinical case in Colombia.

BACKGROUND: Mainly affecting poorer populations in remote (sub)tropical areas, paragonimiasis is considered one of the world's most neglected tropical diseases by the World Health Organization. It is usually acquired by consuming undercooked freshwater crustaceans and primarily affects the lungs, but may ectopically migrate to other organs to produce a multisystemic clinical presentation. This study details what appears to be the first documented case in South America and particularly in Colombia of cerebral paragonimiasis and infection by the crab Moreirocarcinus emarginatus. CASE PRESENTATION: After consuming this crab, a 32-year-old Colombian male developed dyspnea and headache. A chest X-ray revealed tension pneumothorax (TPT) and a thoracoscopic lung biopsy bronchiolitis, eosinophilia, and granulomatous reaction due to Paragonimus parasites. RESULTS: Brain tomography and MRI also showed a configuration typical of Paragonimus parasites, namely a predominantly cystic area near the left cuneus with diffusion restriction, a hemosiderin halo, and linear enhancement characteristic of the tunnel sign, indicating an adult worm moving in the brain. CONCLUSIONS: Cerebral paragonimiasis can occur in the Colombian population and Moreirocarcinus emarginatus can be its cause. In rural areas, education about raw crab consumption would be beneficial, and physicians addressing lung-related complaints should bear paragonimiasis in mind and ask about raw crab consumption.

Hypolobocera guayaquilensis (Decapoda: Pseudothelphusidae): A New Crab Intermediate Host of Paragonimus mexicanus in Manabí Province, Ecuador.

To determine that Paragonimus sp. is actively transmitted in a tropical area of the Pacific region of Ecuador where human cases of pulmonary paragonimiasis have recently been documented, a total of 75 freshwater crabs were collected from 2 different streams in the Pedernales area of Manabí Province, Ecuador. All collected crabs were identified as Hypolobocera guayaquilensis based on morphological characteristics of the male gonopods. The hepatopancreas of each crab was examined by compressing it between 2 glass plates followed by observation under a stereomicroscope. Excysted Paragonimus metacercariae were detected in 39 (52.0%) crabs and their densities varied from 1 to 32 per infected crab. There was a positive relationship between crab size and metacercarial density. Sequences of the second internal transcribed spacer region of the ribosomal RNA gene of the Paragonimus metacercariae obtained in this study were identical to those of Paragonimus mexicanus deposited in the DDBJ/EMBL/GenBank database. Thus, the present study is the first to confirm that the crab species H. guayaquilensis is the second intermediate host of P. mexicanus in Manabí Province, Ecuador. Because this crab might be the possible source of human infections in this area, residents should pay attention to improper crab-eating habits related with a neglected parasitic disease, i.e., paragonimiasis.

Morphological and molecular characterization of Paragonimus caliensis Little, 1968 (Trematoda: Paragonimidae) from Medellin and Pichinde, Colombia.

Paragonimiasis is a subacute to chronic inflammatory granulomatous lung disease caused by the genus Paragonimus. In Latin America Paragonimus mexicanus Miyazaki & Ishii, 1968 is the only confirmed species to cause human infections. Paragonimus caliensis Little, 1968 is an uncommon species often regarded as a synonym of P. mexicanus. Recently, the study of two types of Paragonimus metacercariae from Costa Rica has provided new molecular and morphological evidence that P. caliensis is a separate species from P. mexicanus. In the present study, molecular, morphological and phylogenetic tools have been used to characterize two populations of Paragonimus located at west of Medellin, Antioquia and at Pichinde, Valle del Cauca (type locality of P. caliensis), Colombia. Adults and metacercariae obtained from Medellin, and metacercariae from Pichinde were analyzed. For morphological observations we used light microscopy and scanning electron microscopy (SEM). Morphology of metacercariae and adults matched with the holotype of P. caliensis. The number and arrangement of sensory papillae in the acetabulum region differs from the morphotypes reported for P. caliensis in Costa Rica. Two morphotypes in branching patterns of ovary and two morphotypes in branching patterns of testes were identified. The main morphological differences between P. caliensis and P. mexicanus corresponded to the size of gonads and their relative positions in the body, and the occasional presence of a cyst wall in P. caliensis metacercariae. The molecular and phylogenetic analyses (using nuclear ribosomal ITS2 and partial cytochrome c oxidase subunit 1 CO1 sequences) confirmed that P. caliensis from the type locality is the same species from Medellin and Costa Rica. Furthermore, these analyses also suggest genetic as well as geographical separation of P. caliensis populations between Colombia and Costa Rica. Currently, P. mexicanus and P. caliensis are sympatric in the Colombian Pacific bioregion, and specific diagnosis based on their egg size is not possible. Therefore, it is necessary to determine the biogeographic distribution ranges of both species and to implement molecular techniques to establish the role of P. caliensis in human paragonimiasis in Colombia.

Morphological and molecular characterization of the metacercaria of Paragonimus caliensis, as a separate species from P. mexicanus in Costa Rica.

The trematode Paragonimus mexicanus is the etiological agent of paragonimiasis, a food-borne zoonotic disease in Latin America. This species, as well as Paragonimus caliensis, have been reported from Costa Rica, but it is not known if the two are synonymous. Two types of Paragonimus metacercariae from freshwater pseudothelphusid crabs from several localities in Costa Rica were recognized by light microscopy. Morphologically, these corresponded to descriptions of P. mexicanus and P. caliensis. Metacercariae of the former species lacked a membrane or cyst and their bodies were yellow in color. Those of P. caliensis were contained in a transparent thin cyst and were pink in color. Morphotypes of metacercariae were determined using scanning electron microscopy (SEM). Based on the number and distribution of papillae in the ventral sucker, three morphotypes were found for P. mexicanus and two for P. caliensis. Analysis of DNA sequences (nuclear ribosomal 28S and ITS2 genes, and partial mitochondrial cox1 gene) confirmed the presence of P. mexicanus and provided the first molecular data for P. caliensis. The two species are phylogenetically distinct from each other and distant from the Asian species. The confirmation of P. caliensis as a separate species from P. mexicanus raises several questions about the ecology, biological diversity, and epidemiology of the genus Paragonimus in Costa Rica.

[Paragonimus sp. in crabs and awareness of the educational community to aquatic ecosystems in La Miel and La Clara, Caldas, Antioquia]. / Paragonimus sp. en cangrejos y sensibilización de la comunidad educativa hacia los ecosistemas acuáticos de La Miel y La Clara, Caldas, Antioquia.

INTRODUCTION: Previous research has demonstrated the presence of Paragonimus spp. in crabs of three municipalities located in the Medellín river basin. To design a plan for the control and prevention of paragonimosis, parasite presence must be demonstrated in the upper area of the river, and an awareness campaign of its risks must involve the participation of the educational community in focusing attention on the local aquatic resources. OBJECTIVE: The presence of Paragonimus spp. metarcercariae was established in freshwater crabs from La Miel and La Clara streams, and an awareness campaign created public awareness to management of the aquatic ecosystems for risk reduction. MATERIALS AND METHODS: Nine crabs were collected from streams of La Miel and La Clara villages in 2007 and 2008. The crabs were indentified, sacrificed in the laboratory and examined for presence of Paragonimus spp. metacercariae. Five workshops were conducted with students of educational institutions of the two villages to improve the student perception of paragonimosis risks associated with the local streams. RESULTS: The crabs were identified as Hypolobocera bouvieri. One crab was found with Paragonimus spp. in La Miel. A mollusk, Aroapyrgus colombiensis, another host of the parasite was found in La Clara. That aquatic ecosystems are a great attraction for the local was well established. CONCLUSIONS: Foci of Paragonimus spp. flatworms are present in the upper area of Medellín river which includes Caldas. However, local school children were uninformed about the risk of paragonimosis associated with the local aquatic ecosystem. Therefore this province must be included in programs for prevention and control of paragonimosis. The prevalence of this disease recommends educational programs that promote awareness of local aquatic ecosystems and their parasitic inhabitants.

Morphological description and life cycle of Paragonimus sp. (Trematoda: Troglotrematidae): causal agent of human paragonimiasis in Colombia.

The first morphological description is made of all stages of the life cycle of a Paragonimus species infecting humans in Colombia. Larval stages were obtained both in vitro and from field collections. Adult Paragonimus spp. are described. The aquatic snail Aroapyrgus sp. serves as an intermediate host of this species, both naturally and experimentally, yielding rediae and cercariae. Crabs (Hypolobocera bouvieri monticola and H. emberarum) were found to be the natural second intermediate hosts, and individuals of another crab species (Strengeria sp.) were also infected in the laboratory.

Comparison of two single-day regimens of triclabendazole for the treatment of human pulmonary paragonimiasis.

A well-tolerated single-day treatment regimen is required for the community-based treatment of pulmonary paragonimiasis. To identify an optimal single-day treatment regimen, an open clinical trial was performed to compare the efficacy of 2 single-day regimens of triclabendazole. From 1991 to 1993, a total of 154 Ecuadorian patients were allocated to receive either a single dose or 2 doses of 10 mg/kg triclabendazole. Treatment with both regimens was associated with a rapid parasitological response (determined by clearance of Paragonimus eggs from sputum), resolution of most clinical symptoms and radiological signs, and only mild adverse effects. At 3 months after treatment, the cure rate (clearance of eggs from sputum) was 90.9% in the 2-dose group and 84.4% in the single-dose group. Re-treatment with a second 2-dose regimen was curative in all remaining patients by 1 year of follow-up. Our findings show that both single-day treatment regimens of triclabendazole were highly effective in the treatment of pulmonary paragonimiasis and suggest that patients should be re-examined at 3 months to determine the need for repeat treatment.

Paragonimiasis: a view from Columbia.

Paragonimiasis is a zoonosis caused by adult trematodes of the Paragonimus genus. The infection in humans is a result of a complex transmission cycle that includes two obligate intermediate hosts, a snail and a crustacean or a crayfish, and a definitive mammalian host. It has been shown that 9 of the more than 40 species of Paragonimus described affect humans in over 39 countries in Asia, Africa and America. It is estimated that 20.7 million people have paragonimiasis and it is calculated that 195 million people are at risk of being infected. The illness usually is caused once the parasite has settled in the lung at the site of the main clinical symptoms: cough, thoracic pain and hemoptysis. The diagnosis of paragonimiasis is based on the patient's history, the parasitological findings (ova in sputum and in feces), and the result of radiological and immunological tests. In severe cases, the patient may suffer from life-threatening hemoptysis or pneumothorax. Currently, praziquantel is the drug of choice.

Paragonimosis in the Cajabamba and Condebamba districts, Cajamarca, Peru.

Stool samples from 409 pre-school and school students, living in six villages of the Cajabamba and Condebamba districts, Cajamarca, Perú, were examined using wet preparations and Lumbreras' method, looking for Paragonimus eggs. Fecal and sputum samples from two children (0. 5%) of 6 and 8 year-old showed eggs of Paragonimus. One hundred and twenty freshwater crabs, Hypolobocera chilensis eigenmanni, were collected from the Condebamba valley and 21 (17.5%) of them were infected with P. mexicanus (syn. P. peruvianus) metacercariae. Our results show the persistence of Paragonimus in human beings and in the main source of infection, the crabs.

Paragonimosis in the Cajabamba and Condebamba districts, Cajamarca, Peru

Stool samples from 409 pre-school and school students, living in six villages of the Cajabamba and Condebamba districts, Cajamarca, Perú, were examined using wet preparations and Lumbreras' method, looking for Paragonimus eggs. Fecal and sputum samples from two children (0.5 percent) of 6 and 8 year-old showed eggs of Paragonimus. One hundred and twenty freshwater crabs, Hypolobocera chilensis eigenmanni, were collected from the Condebamba valley and 21 (17.5 percent) of them were infected with P. mexicanus (syn. P. peruvianus) metacercariae. Our results show the persistence of Paragonimus in human beings and in the main source of infection, the crabs

Paragonimiasis pulmonar, cerebral y abdominal. Presentación de un caso de revisión de la enfermedad en Colombia / Pulmonary, cerebral and pelvic pargonimiasis. Prersentation of one case and review in Colombia

Presentamos el caso de un paciente con compromiso pulmonar, cerebral y pélvico cuyo estudio confirmó finalmente el diagnóstico de una paragonimiasis. Esta es una zoonosis (enfermedad del hombre y de los animales) causada por parásitos tremátodos del género paragonimus, que afecta pricipalmente el pulmón y otros órganos como el cerebro, el páncreas y el hígado. El parásito causante tiene un ciclo de vida que involucra moluscos y crustáceos como los cangrejos. Presenta una distribución geográfica particular en Asia y Sur América. Se había descrito desde hace muchos años en países vecinos a Colombia pero solamente en forma reciente se han publicado casos y series de casosnacionales con pacientes provenientes del Meta y de la comunidad Emberá. El caso que presentamo sirve de ilustración y de señal de alerta para considerar esta parasitosis en el diagnóstico diferencial de pacientes con enfermedad pulmonar, eosinofilia y procedencia de regiones como Chocó, Meta o Guaviare

Paragonimiasis de presentación atípica: en un paciente pediátrico: revisión de la literatura

La paragonimiasis es una afección parasitaria que clasicamente compromete el parénquima pulmonar provocando la presencia de algunos patrones radiológicos, que aunque poco específicos deben sospecharse en sitios donde la enfermedad es endémica como en nuestro país. Es un problema clínico radiológico cuando el parásito ocupa sitios poco frecuentes como la cavidad pleural. Presentamos el caso de un paciente con localización errática en la cabidad pleural a manera de empiema que es la forma clínica más rara conocida (1 al 2por ciento).

Paragonimiasis pulmonar: informe de un caso / Pulmonary paragonimiasis. A report of an isolated case

La paragonimiasis es una enfermedad parasitaria causada por un tremátodo del género Paragonímus. Es endémica en Corea, Japón y China. En América Latina se han identificado algunos casos. En México hay 6 casos en la literatura revisada. El caso que nos ocupa es el de mujer de 41 años que ingresó al Instituto Nacional de Enfermedades Repiratorias con tos crónica, hemoptisis y pérdida de peso, a la que se practicó lobectomía superior derecha. En el estudio histopatológico se observaron lesiones granulomatosas que contenían huevecillos de Paragonimus

Pathological and parasitological aspects of the first autochthonous case of human paragonimiasis in Venezuela.

A worm found in histopathologic sections of a lung piece of a young Venezuelan male was identified as a Paragonimus sp. Definitive identification of the species was not possible since only a deteriorated segment of the worm was recovered, nevertheless comparison with other known species is discussed. This is the first report of an indigenous case of human paragonimiasis in Venezuela.

Infeccion natural de Ptychophallus tristani (Crustacea: Decapoda) con metacercarias de paragonimus mexicanus (Trematoda) en Tabarcia de Mora, Costa Rica / Natural infection of Ptychophallus tristani (Crustacea: Decadopa) by metacercariae of Paragonimus mexicanus (Trematoda) in Tabarcia de Mora, Costa Rica

Se determinó el grado de infección de los crustáceos Ptychophallus tristani presentes en una área endémica de Paragonimus mexicanus, encontrándose que de un total de 182 cangrejos, 161 resultaron positivos (88,5%) por metacercarias. Al comparar la infección entre machos y hembras no se encontró diferencias estadísticamente significativas y en relación a su tamaño se observó una relación directa entre tamaño y porcentaje y grado de infección. Se estableció que no existen diferencias significativas en relación a la época del año en que los cangrejos fueron capturados lo que epidemiológicamente es importante pues indica que las fuentes de infección permanecen potencialmente activas durante todo el año