Ana Espert

The comparative development of Echinostoma caproni (Trematoda: Echinostomatidae) adults in experimentally infected hamsters and rats

The aim of the present study was to compare the development of Echinostoma caproni (Trematoda: Echinostomatidae) adults in two host species displaying different degrees of compatibility with this parasite. For this purpose, the variability in the worm recovery, egg output, and morphology of E. caproni adults during the course of experimental infections in hamsters and rats was analyzed. Student’s t-tests and two-factor ANOVA analysis with the time post-infection and the host species as independent variables, and Bonferroni t-tests as post hoc analysis were used for the study. Worm recovery and egg output were host species dependent. The values were significantly higher in the worms established in hamsters than those in rats. The oral sucker area, pre-pharynx length, and pharynx area were the most conservative features, and no significant variability related to the host species was detected. In contrast, body area, collar width, esophagus length, cirrus sac area, ventral sucker area, ovarian area and anterior and posterior testicular areas were significantly higher in those worms collected from hamsters. Moreover, significant worm age-host species interactions were found for body area, ovarian area, ventral sucker area, and anterior and posterior testicular areas.

An experimental study of the reproductive success of Echinostoma friedi (Trematoda : Echinostomatidae) in the golden hamster.

Viable eggs produced weekly per infective stage was used as a measure of the reproductive success of Echinostoma friedi during the first 12 weeks of infection in hamsters. The weekly reproductive success was not constant during the experiment in relation to the egg output and the proportion of viable eggs produced. The egg release started during week 2 post-inoculation, attaining a maximum during week 3. A decline in egg output was observed from week 9. Viable eggs were only produced from week 3 post-inoculation and a maximum was attained at week 4 of the experiment. A decline in egg viability was observed from week 9. Considering together the egg output and the egg viability, the maximum weekly reproductive success was obtained during week 4 post-inoculation. The changes in the weekly reproductive success were not reflected in variations in worm numbers and body sizes during the course of the infection. The humoral immune response of golden hamsters during the infection with E. friedi was determined. Increases of IgG levels against somatic and excretory/secretory products of E. friedi were detected coinciding with the reduction in the reproductive success.

DEVELOPMENT OF AN ANTIBODY-BASED CAPTURE ENZYME-LINKED IMMUNOSORBENT ASSAY FOR DETECTING ECHINOSTOMA CAPRONI (TREMATODA) IN EXPERIMENTALLY INFECTED RATS: KINETICS OF COPROANTIGEN EXCRETION

The present study reports on the development of a coproantigen capture enzyme-linked immunosorbent assay (ELISA) for detecting Echinostoma caproni in experimentally infected rats. The capture ELISA was based on polyclonal rabbit antibodies that recognize excretory–secretory (ES) antigens. The detection limit of pure ES was 3 ng/ml in sample buffer and 60 ng/ml in fecal samples. The test was evaluated using a follow-up of 10 rats experimentally infected with 100 metacercariae of E. caproni, and the results were compared with those of other diagnostic methods such as parasitological examination and antibody titers determined by indirect ELISA. Coproantigens were detected in all the infected rats from the first day postinfection (DPI). The period of maximal coproantigen excretion was between 7 and 21 DPI. The values remained positive until 49–56 DPI, coinciding with the disappearance of the eggs in the stool samples of the infected rats. The kinetics of coproantigen detection were correlated with those of egg output. The present assay provides an alternative tool for the diagnosis of the echinostome infections. The proposed capture ELISA makes possible an earlier diagnosis than that provided by parasitological examination and indirect ELISA and also allows for the differentiation of past and current infections. Our results show that this assay can also be used to monitor the course of echinostome infections.

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.

KINETICS OF ECHINOSTOMA CAPRONI (TREMATODA: ECHINOSTOMATIDAE) ANTIGENS IN FECES AND SERUM OF EXPERIMENTALLY INFECTED HAMSTERS AND RATS

This study reports on the kinetics of antibody production to Echinostoma caproni and the dynamics of antigens in feces and sera in 2 experimental hosts (hamsters and rats) that display different degrees of susceptibility with this echinostome. Echinostoma caproni produced chronic infections in hamsters, whereas rats lost the infection at 49–56 days postinfection (DPI). Hamsters developed higher antibody responses than rats, probably in relation to different intestinal absorptions of worm antigens in each host species. The levels of coproantigens were indicative of the course of infection in each host. Positive coproantigen levels were detected at 1–2 DPI in both hosts, and the values remained positive until the end of the experiment in hamsters; in rats, the coproantigen levels reverted to negative values, coinciding with the loss of infection. High levels of circulating antigens were detected in hamsters from 21 DPI to the end of the study. In contrast, low levels of E. caproni seroantigens were detected in rats only. These observations may reflect the differences in local inflammatory responses induced by E. caproni in each host species.

A QUANTITATIVE APPROACH TO THE EXPERIMENTAL TRANSMISSION SUCCESS OF ECHINOSTOMA FRIEDI (TREMATODA: ECHINOSTOMATIDAE) IN RATS

Using a range of parameters, the ability of rats (Rattus norvegicus) to successfully transmit Echinostoma friedi to the next host was examined under experimental conditions. The concept of Experimental Transmission Success (TM), defined as the number of hosts that become successfully infected after exposure to a number of infective stages produced by a previous host per unit of inoculation at which this latter host was exposed, was introduced. Using data for the egg output and miracidium hatching and infectivity, the TM permits us to estimate the ability of a particular defintive host species to successfully transmit a parasite species. This concept may be also useful to compare the transmission fitness of a parasite in different definitive host species. Moreover, variations of the Experimental Transmission Success over the course of the infection were calculated by the use of the Weekly Experimental Transmission Success (TMW). Overall, considering the complete duration of the experiment, the TM of E. friedi using rats as definitive hosts was 0.68 infected snails/metacercaria. However, positive values of the TMW were only obtained from 2 to 4 wk post-infection, with a maximum during the third wk post-infection. When comparing the TM values of E. friedi in rats with those calculated in hamsters on the basis of previously published data, E. friedi appears to be more appropriate to move through this portion of its life cycle when using hamsters (Mesocricetus auratus) as the final host than rats.

Immunological Characterization of Somatic and Excretory–Secretory Antigens of Echinostoma caproni (Trematoda: Echinostomatidae) in Experimentally Infected Rats

Abstract The antigenic properties of somatic (SoP) and excretory–secretory products (ESP) of Echinostoma caproni were studied in experimentally infected rats. Sodium dodecyl sulphate–polyacrylamide gel electrophoresis demonstrated a total of 15 and 9 major polypeptides, in the molecular weight range 10–165 kDa, in SoP and ESP products, respectively. Further analysis revealed some common as well as several unique polypeptides for each type of extract. A total of 2 polypeptides, weighing 58 and 115 kDa, were found in both types of antigen. Indirect enzyme-linked immunosorbent assay analysis using both types of antigens showed that rats developed a weak and slow response in infections with E. caproni. However, the response seems to be mainly directed against excretory–secretory antigens. We have also defined by Western blot 11 and 7 major antigenic polypeptides in SoP and ESP, respectively, that were recognized by sera from experimentally infected rats. Only 2 of these polypeptides were common to both types of antigens. A total of 9 antigenic polypeptides, ranging from 24 to 137 kDa, were found to be characteristic of SoP, whereas 5 polypeptides, weighing from 28 to 117 kDa, were characteristic of ESP. The kinetics of antibody production against discrete polypeptides composing both types of antigens were determined during the course of the infection.

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.