Soil-transmitted Helminth Infections in Schoolchildren of Laguna de Perlas (Nicaragua).

Soil-transmitted helminth (STH) prevalence, intensity, polyparasitism and co-infections in 425 children from 3 schools of Laguna de Perlas (Nicaragua) were investigated. Single stool samples were analysed by the formalin-ether method and the Kato-Katz. A total of 402 (94.6%) children were infected. Trichuris trichiura 308 (72.4%), Ascaris lumbricoides 115 (27.1%) and Hookworms 54 (12.7%) were the most prevalent STHs. Polyparasitism (322; 75.8%) with two species was most prevalent (109; 25.6%). T. trichiura with A. lumbricoides (19.3%) and T. trichiura with Hookworm (6.8%) were the most common combinations. Positive associations were observed between T. trichiura and A. lumbricoides and T. trichiura and Hookworm (p = 0.0001). Highest Hookworm intensities appeared when three STH co-infections occurred. Moderate- heavy STH intensities appear in up to 42.1% in trichuriasis, 57.5% in ascariasis and 11.1% in Hookworm infections. Integrated control interventions covering children need to be implemented mainly in a rural environment.

Partial resistance to homologous challenge infections of the digenean Echinostoma caproni in ICR mice.

In the present paper, we analyse the effect of a primary infection of ICR mice with Echinostoma caproni (Trematoda: Echinostomatidae) on the generation of resistance against homologous challenge infections. In ICR mice, E. caproni induces chronic infections concomitantly with strong responses characterized by the development of T-helper 1 (Th1)-type local immune responses with elevated levels of local interferon-gamma (IFN-γ) and inflammatory and antibody responses. Here, the effect of the response generated against a primary infection with E. caproni in the generation of resistance against subsequent homologous infections was analysed. For this purpose, ICR mice were challenged with metacercariae of E. caproni and the results obtained showed that primary infection induces partial resistance against subsequent homologous infections in ICR mice. This resistance was expressed as a reduced rate of infection, worm recovery and worm size, indicating that primary infection induces changes in the host, making a hostile environment for the development of the parasite.

Echinostoma caproni (Trematoda): differential in vivo mucin expression and glycosylation in high- and low-compatible hosts.

Enhanced mucus production and release appears to be a common mechanism for the clearance of intestinal helminths, and this expulsion is normally mediated by Th2-type immune responses. To investigate the factors determining the expulsion of intestinal helminths, we have analysed in vivo expression of mucin genes at the site of infection in two host species displaying different compatibility with Echinostoma caproni (Trematoda). Surprisingly, a general down-regulation on mucin mRNA expression was detected in low-compatible hosts (rats) coinciding with the development of Th2/Th17 responses and the early rejection of the worms from the intestinal lumen. This suggests the existence of a mechanism by which the parasites can modulate the mucus barrier to favour their survival. In highly compatible hosts (mice), some mucin genes were found to be up-regulated throughout the infection, probably, to protect the intestinal epithelium against the infection-induced inflammation developed in this host species. Moreover, infection-induced changes on mucin glycans were also studied by lectin histochemistry. Similar alterations were detected in the ileum of infected mice and rats, except with SNA lectin, indicating that sylated mucins might play an important role in determining the evolution of the infection in each host species.

Haematological changes in the laboratory rat Rattus norvegicus infected with Echinostoma caproni (Trematoda: Echinostomatidae).

To study possible indirect effects of the infection with intestinal helminths, 12 Rattus norvegicus (Wistar) were each experimentally exposed to 100 metacercariae of Echinostoma caproni, and blood samples were taken weekly up to 4 weeks post-exposure for comparison with control rats. Values of haematocrit (HCT), red blood cells (RBC), platelets (PLT), white blood cells (WBC), haemoglobin (HGB) and haematimatrix indices, and mean corpuscular haemoglobin concentrations (MCHC) were determined. In addition, leucocyte counts, including lymphocytes, neutrophils, monocytes, eosinophils and basophils were analysed. These parameters, including the leucocyte counts, showed no significant differences, except for MCHC at 4 weeks post-exposure. The present results indicate that in rats infected with E. caproni, although eosinophilia did not significantly increase, a significant reduction in MCHC was associated with an increase in the number of RBC.

Human infection by a "fish tapeworm", Diphyllobothrium latum, in a non-endemic country.

We document a case of locally acquired "broad- or fish-tapeworm" infection caused by Diphyllobothrium latum in a 27-year-old Spanish man, confirmed by molecular analysis (COI gene). The patient had naturally expelled a worm of 110 cm in length, but the physical examination did not yield any remarkable findings, and the patient did not suffer from any particular symptoms. Laboratory test results were normal except for a remarkable increase in the red blood cell count, and an evident decrease in the mean corpuscular volume and mean corpuscular hemoglobin. Vitamin B12 and folic acid values were in the normal range without signs of anemia. It was suggested that these anomalies in erythrocyte formation might not be related to the parasite, and analysis of the patient's anamnestic data revealed that the infection could only have been caused by the ingestion of imported fish, although no light could be shed on the specific source of infection. From a public health viewpoint, this human case of fish-borne zoonosis is exemplary, suggesting that not only is control of fish and fish product quality essential, but also increased awareness of the general population with regard to changes in culinary habits.

The influence of adult worm age and definitive host on the transmission of Echinostoma caproni: egg hatchability and miracidial infectivity.

We analysed the influence of different ages of Echinostoma caproni adult worms reared in different definitive host-species on egg hatchability and miracidial infectivity. Eggs obtained from 2-, 4- and 6-week-old adult worms collected from rats, hamsters and mice, hatched miracidia that were used to infect laboratory-reared specimens of Biomphalaria glabrata. Highest egg hatchability was obtained with eggs derived from hamsters and mice. Egg hatchability did not seem to be affected by the age of E. caproni adult worms. Miracidial infectivity was not affected by the definitive host species. However, miracidial infectivity was affected by the age of adult worms when they developed in the mice intestine. The long period of miracidial infectivity in hamsters revealed this definitive host as the most suitable for experimental development of the life cycle and parasite transmission.

Proteomic analysis of Strongyloides stercoralis L3 larvae.

Strongyloidiasis can be perpetuated by autoinfection with the filariform larvae L3, causing asymptomatic chronic infections and creating a population of carriers, affecting not only developing countries. So far, very little is known about the proteins that interact with the human host, and few proteins from the infective Strongyloides stercoralis L3 have been characterized. Here, we report results obtained from a proteomic analysis of the proteins from S. stercoralis L3 larvae obtained from patients. Since the genome of S. stercoralis is not yet available, we used proteomic analysis to identify 26 different proteins, 13 of them released by short digestion with trypsin, which could represent surface-associated proteins. The present work extends our knowledge of host-parasite interactions by identifying proteins that could be of interest in the development of diagnostic tools, vaccines, or treatments for a neglected disease like strongyloidiasis.

Sympatric and allopatric experimental infections of the planorbid snail Gyraulus chinensis with miracidia of Euparyphium albuferensis (Trematoda: Echinostomatidae).

An experimental infection with echinostomatid miracidia in sympatric or 'local' vs. allopatric or 'away' snail combinations, as a model to examine parasite compatibility, was carried out. We employed Euparyphium albuferensis miracidia to infect Gyraulus chinensis snails, from three different natural parks: Albufera (Valencia, Spain); the Ebro Delta (Tarragona, Spain) and Coto de Doñana (Huelva, Spain). Insignificant differences between the three snail strains were noted for the infection rate and the rhythm of daily cercarial production. However, a significantly higher total cercarial production per snail, patent period and life span were observed in local snails. The different infection characteristics in the three G. chinensis strains considered reveal that E. albuferensis miracidia demonstrate local adaptation.

Kinetics of antibodies and antigens in serum of mice experimentally infected with Echinostoma caproni (Trematoda: Echinostomatidae).

The present study reports on the kinetics of antibodies and antigens in serum of mice experimentally infected with 75 metacercariae of Echinostoma caproni during the first 12 wk postinfection (wpi). Antibody titers in the serum of mice were determined by an indirect enzyme-linked immunosorbent assay (ELISA) using excretory/secretory (ES) antigens of E. caproni. The early detection of antibodies against ES antigens of E. caproni is feasible using indirect ELISA. Mice developed significant antibody responses at 2 wpi, and the values progressively increased until the end of the experiment. This may be related to the intestinal absorption of adult worm antigens that induces humoral responses. The presence of E. caproni circulating antigens was determined by a capture ELISA based on polyclonal rabbit antibodies against ES antigens of E. caproni. High levels of seroantigens in mice were detected by 1-2 wpi, probably because of the local inflammatory responses in mice induced by the adult worms. A drop in circulating antigen levels was observed at 9 wpi, which could reflect changes in the intestinal tissues over the course of the infection.

Production and chronobiology of emergence of the cercariae of Euparyphium albuferensis (Trematoda: Echinostomatidae).

The production and the chronobiology of emergence of the cercariae of Euparyphium albuferensis from Gyraulus chinensis experimentally infected with a single miracidium were established during 28 consecutive days from the first day of cercarial shedding. Moreover, the effect of a sudden change in light-dark cycling was investigated. Although the daily cercarial shedding rates show great variability, a progressive increase in cercarial production was observed in the first weeks of the cercarial shedding periods, probably in relation to the demography of intramolluscan larval stages. Under 12:12 light-dark cycling conditions, E. albuferensis cercariae emerged in the light, and the rhythm was circadian. The sudden change in the light-dark cycle resulted in corresponding alterations in emergence patterns, which shows that cercarial emergence was correlated to light-dark alternation. The ecological consequences of daily emergence by mobile cercariae whose target hosts are organisms that regularly occur in the same habitat with the molluscan emitting host, such as those of E. albuferensis, are discussed with reference to the hypotheses proposed to date, and an alternative hypothesis is proposed.

SSU rDNA characterization of lymnaeid snails transmitting human fascioliasis in South and Central America.

The small subunit (18S) rRNA gene sequences of the lymnaeid morphs I and II (Mollusca: Gastropoda: Basommatophora: Lymnaeidae) transmitting human fascioliasis in the high endemic zone of the northern Bolivian Altiplano and of Lymnaea cubensis from Mexico and Guadeloupe island (Caribbean) have been obtained by direct polymerase chain reaction PCR cycle sequencing and silver staining methods and compared to that of the 6 most common European Lymnaeidae species. Results allow us to establish definitively the distinction between the lymnaeids from the northern Bolivian Altiplano and L. cubensis. Lymnaea cubensis is a valid species distributed in North and Central America but absent in the northern Bolivian Altiplano. Lymnaeid morphs I and II from the northern Bolivian Altiplano both present the same 18S rDNA sequence, which is moreover identical to that of the European species Lymnaea truncatula. Significant nucleotide substitutions in helix E10-1 of the variable region V2 of the secondary structure suggest the need for distinguishing L. cubensis in the subgenus Lymnaea (Bakerilymnaea) with L. (B.) cubensis as type species. The subgenus Lymnaea (Fossaria) is retained, with L. (F.) truncatula as type species. The larger number of nucleotides in the 18S rDNA sequence of L. cubensis (1,860 bp) with regard to the other Lymnaea species (1,843-1,852 bp) is tentatively related to the more ancient paleogeographic origin of L. cubensis. The grouping of L. cubensis with L. truncatula and the relationship of Lymnaea auricularia with Lymnaea peregra in the phylogenetic trees obtained show an evolutionary parallelism with the digenean parasite species they transmit, namely Fasciola hepatica and Fasciola gigantica, respectively.

Prevalence and risk factors related to intestinal parasites among children in Department of Rio San Juan, Nicaragua.

BACKGROUND: The present study was carried out to assess the prevalence of intestinal parasite infections among children in Department of Rio San Juan (Nicaragua), to explore the extent of polyparasitism, and to identify the risk factors that might favour transmission of these parasites. METHODS: A coprological study of single stool specimen, collected at random from 382 children (167 boys and 215 girls) aged 2-15 years from two municipalities, and a questionnaire, concerning demographic and epidemiological data on sanitary and behavioural habits, was filled in to identify risk factors. RESULTS: About 93% of the children presented infection. Blastocystis hominis, the most frequently protozoa, and Trichuris trichiura, Ascaris lumbricoides and hookworm, the most prevalent helminths. Polyparasitism (85.4%[326/382]), with the highest percentages detected in females (p=0.001), in the 6-11 year age group (p<0.001) and having a rural background (p<0.001). A firm relationship between any parasite and age (OR=1.2, p=0.036) and rural background (OR=0.3, p=0.027) was detected. Living in rural areas and drinking river water were also positively associated with each of the STH species detected, and walking barefoot was also associated with hookworm infection. CONCLUSIONS: Government efforts should be focused on controlling the risk factors associated with these enteroparasites, with health education programmes in rural areas of Nicaragua.

Prevalence and molecular characterization of Cryptosporidium in schoolchildren from department of Rio San Juan (Nicaragua).

A cross-sectional study of cryptosporidiosis was carried out in Nicaragua. The prevalence of Cryptosporidium infection was determined in 272 (110 boys and 162 girls) schoolchildren, aged between 4 to 15 years from department of Rio San Juan. The total percentage obtained for Cryptosporidium (35.7%) was one of the highest reported so far. Cryptosporidium appeared in 94.8% of multiparasitism cases. No significant statistical differences were detected in the cryptosporidiosis prevalence between loose/watery (22.2%) and soft/formed (36.7%) stool samples. No significant age and sex differences were observed. This is the first report to identify Cryptosporidium in Nicaragua at species level, providing a preliminary molecular characterization of all positive samples, such as Cryptosporidium parvum (genotype 2). The high prevalence of C. parvum suggests that animals may be potential sources of infection for human cryptosporidiosis, although C. parvum infections may have originated from humans themselves. The human health problem caused by Cryptosporidium in this region may be related to the poor human hygiene/sanitation and contamination of the environment, food, or water supplies. Continuous exposure to the parasite could have been protective against development of symptoms in the children examined.

Infectivity of Echinostoma friedi miracidia to different snail species under experimental conditions.

The infectivity of Echinostoma friedi (Trematoda: Echinostomatidae) miracidia was studied experimentally in a range of laboratory-reared snails that coexist in the same natural locality, namely Radix peregra, Lymnaea fuscus, L. truncatula (Lymnaeidae), Gyraulus chinensis, Helisoma duryi (Planorbidae) and Physella acuta (Physidae), and snails from different geographical origins acting naturally or experimentally as intermediate hosts of Schistosoma spp., namely Planorbarius metidjensis (from Málaga, Spain), Biomphalaria glabrata (Guadeloupe), B. alexandrina (Egypt) (Planorbidae), Bulinus cernicus (Mauritius), B. globosus (Zambia), B. natalensis (South Africa) and B. truncatus (Niger) (Bulinidae). Six species of snails were found to be susceptible, with the rate of infection ranging from 0 to 36.7%. The highest infection was detected in R. peregra. The low host specificity of E. friedi might have an epidemiological significance as a requisite for a recent establishment in a new geographical area.

The life cycle and transmission dynamics of the larval stages of Hypoderaeum conoideum.

The morphology of the different larval stages and life cycle of Hypoderaeum conoideum (Trematoda: Echinostomatidae) are described. The freshwater snail species Lymnaea peregra (Gastropoda: Lymnaeidae) serves as the natural first intermediate host and this and L. corvus serve as experimental first intermediate hosts. These and other freshwater snails, such as Physella acuta and Gyraulus chinensis, in turn serve as second intermediate hosts. Adult worms were obtained from chicks and ducks, but not from rats, mice and golden hamsters. The morphology of the larval stages is compared with previous work on H. conoideum. Several aspects of the biology of the life history stages are described with emphasis on the transmission dynamics of the free-living stages. Differential suitability of the snail species that may act as first and/or second intermediate hosts is studied and discussed.

The life-cycle of Echinostoma friedi n. sp. (Trematoda: Echinostomatidae) in Spain and a discussion on the relationships within the 'revolutum' group based on cercarial chaetotaxy.

The morphology of the different stages and life-cycle of Echinostomna friedi n. sp. are described and figured. The freshwater snail Lymnaea peregra (Gastropoda: Lymnaeidae) serves as the natural and experimental first intermediate host and L. corvus and Gyraulus chinensis (Gastropoda: Planorbidae) as experimental first intermediate hosts. These, and Physella acuta (Gastropoda: Physidae), also serve as second intermediate hosts. Adult worms, possessing 37 collar spines, were obtained from naturally infected Rattus norvegicus and experimentally from albino rats, golden hamsters and chickens. Mice were not suitable experimental definitive hosts. E. friedi differs from the most closely related species in the 'revolutum' group mainly in terms of several morphological and biological features of the life-cycle stages and in its cercarial chaetotaxy. The chaetotaxy patterns of the species of the 'revolutum' group are analyzed and the results show that a taxonomic comparison of these species may be carried out on the basis of the number of sensilla in the clusters CIII VI, CIII V2 (or CIII V1 + CIII V2), CIV DL and UVb. These clusters appear adequate to establish taxonomic relationships between different species within the 'revolutum' group.

Echinostoma friedi: the effect of age of adult worms on the infectivity of miracidia.

The effect of ageing of adults of Echinostoma friedi (Trematoda: Echinostomatidae) on the infectivity of miracidia yielded was analysed. Miracidia were obtained after hatching of eggs obtained from adult worms of E. friedi collected weekly during the course of experimental infections in golden hamsters. Miracidial infectivity, measured in terms of percentage of infection in Lymnaea peregra, was significantly influenced by the age of the adult worms from which the miracidia were derived. Infective miracidia only were obtained from adult worms in the age range from 4 to 9 weeks post-infection. Infectivity was maximal in those miracidia derived from adults collected 8 and 9 weeks post-infection. The results suggest that adult worms producing viable eggs require additional maturation to be able to yield eggs containing infective miracidia.

Survival and infectivity of Hypoderaeum conoideum and Euparyphium albuferensis cercariae under laboratory conditions.

The survival characteristics of the cercariae of Hypoderaeum conoideum and Euparyphium albuferensis (Trematoda: Echinostomatidae) at 20 degrees C and 30 degrees C are described, and the age dependency of their infectivity at 20 degrees C is studied to determine their respective transmission efficiencies. Cercarial survival was found to be age-dependent and was higher at 20 degrees C. For both cercariae, the maximum life-span was 26 h at 20 degrees C and 16 h at 30 degrees C, and their respective times to 50% mortality were similar at each temperature. Both cercariae seem to be well adapted to transmission in their natural habitat, though cercarial infectivity of H. conoideum was higher than that of E. albuferensis, this being correlated with their prevalences in nature. The age-dependency of cercarial survival may be related to steadily diminishing endogenous energy levels, though the delay in attaining maximum infectivity suggests that other factors not related to energy considerations are involved in the delimitation of the cercarial infective period. However, this latter observation may constitute an adaptative mechanism allowing cercarial dissemination.

Miracidial infectivity of Hypoderaeum conoideum (Trematoda: Echinostomatidae): differential susceptibility of two lymnaeid species.

A study was made of the infectivity of Hypoderaeum conoideum miracidia to a range of laboratory-reared specimens of freshwater snail species (Lymnaea peregra, L. corvus, Physella acuta, and Gyraulus chinensis) that coexist with the parasite in the same natural habitat. L. peregra and L. corvus were found to be equally susceptible to the parasite when specimens of each snail species were singly exposed to miracidia. However, when miracidia could choose either lymnaeid species, they showed a high degree of specificity toward L. peregra. The results obtained suggest that H. conoideum miracidia are capable of distinguishing among these lymnaeids in their orientation to the host. This indicates that miracidia might achieve specificity before actually contacting the snail host and suggests that during the host-snail orientation process they respond to signals different from those generated upon snail contact and invasion. The specificity toward L. peregra observed in H. conoideum miracidia seems to indicate adaptation to the snail community in their natural habitat, resulting in enhancement of their transmission.

Larval trematode infections in freshwater gastropods from the Albufera Natural Park in Spain.

Malacological samplings were made from January 1994 to December 1996 in the Albufera Natural Park (Valencia, Spain) to trace the dynamics of molluscan populations and the prevalence and intensity of infection by larval trematodes. A total of 10,533 freshwater gastropods belonging to seven species (Lymnaea auricularia, L. truncatula, L. palustris, L. peregra, Bithynia tentaculata, Physa acuta and Gyraulus chinensis) was examined, and 110 (1.04%) were found to harbour some of the nine distinguishable types of cercariae, namely four echinostome cercariae (Hypoderaeum conoideum, Echinoparyphium recurvatum, Euparyphium albuferensis, and Echinostoma sp.), four furcocercous cercariae, and one xiphidiocercous cercaria. This study shows that the composition of the snail and trematode communities may be determined by the particular environmental conditions present and the human intervention in the area.