A longitudinal survey of gastrointestinal parasites of the black-eared opossum Didelphis aurita at an urban-sylvatic interface area in Southeast Brazil: a morphological and ecological characterization based on helminth eggs.

Parasites are important organisms in ecosystem dynamics and take part in the regulation and structure of host populations. The prevalence, abundance, and distribution of parasites in their host species may be affected by biotic and abiotic factors. Most studies of helminths of Brazilian marsupials are taxonomic descriptions or re-descriptions and records of occurrence. The use of noninvasive techniques for studying parasitic worms of vertebrate hosts is more common in large or threatened species. The aims of this study were to describe and identify the helminth morphotypes and to analyse the parasitological parameters of gastrointestinal helminth eggs obtained in faecal samples of marsupial Didelphis aurita at the Fiocruz Atlantic Forest Biological Station and surroundings, municipality of Rio de Janeiro, southeast Brazil. The common opossums were sampled during a capture-mark-recapture study. Faecal samples collected from each animal were analysed for helminth egg diagnosis using the methods of flotation in sugar solution and sedimentation. Eggs were compared with samples obtained from the uterus of adult worms obtained from a previous study carried out in the same area. Eleven helminth egg morphotypes were found. Among them, seven were identified at the species level. The highest values of parasitic load and prevalence were observed for the families Viannaiidae and Trichuridae. A significant relationship was found between the number of morphotypes and host body size in females. The influence of parasite load on host condition factor or body size was not observed.

Evaluation of changes in the carbohydrate metabolism of Biomphalaria glabrata Say, 1818 exposed to experimental coinfection by Angiostrongylus cantonensis (Nematoda) and Echinostoma paraensei (Trematoda).

The interaction between intermediate snail hosts and helminths can cause metabolic changes in the former. The snails use their reserves for maintenance of their vital processes, by activating the internal defense system and repairing tissue damage, while also supplying necessary energy for the parasites' development. Our aims were to evaluate the lactate dehydrogenase activity and the glucose concentration in the hemolymph of Biomphalaria glabrata experimentally coinfected by Angiostrongylus cantonensis and Echinostoma paraensei. Besides these aspects, the glycogen content in the digestive gland complex and cephalopedal mass along with histochemical changes in parasitized snails were analyzed. The snails were divided in group A (infected by 1200 L1 of A. cantonensis), group E (infected by 20 E. paraensei miracidia), group A + E (co-infected with A. cantonensis first and after a week by E. paraensei), group E + A (co-infected with E. paraensei first and then by A. cantonensis) and control group (not infected). During four weeks after exposure, samples were collected for biochemical and histochemical analyses. In the infected snails, glucose levels and glycogen content in the digestive gland complex and cephalopedal mass were significantly lower, in contrast with an increase of lactate dehydrogenase activity. These results indicate that the intense energy demand resulting from the presence of parasites causes the host snail to accelerate the anaerobic degradation of carbohydrates to obtain energy, in an attempt to maintain homeostasis. Both parasites were observed in histochemical analysis to cause tissue damages in the snails. So, although the snails were able to sustain the coinfection, several metabolic and tissue changes occurred, mainly in those infected with E. paraensei and then with A. cantonensis.

Biological and histological changes in reproductive patterns of Biomphalaria glabrata experimentally co-infected by Echinostoma paraensei and Angiostrongylus cantonensis.

Investigation of co-infection by different helminths in snails can provide essential information about the biology of parasites co-existence under natural conditions. This study aimed to investigate the reproductive and histological changes of Biomphalaria glabrata experimentally co-infected with Echinostoma paraensei and Angiostrongylus cantonensis. Five groups of snails i.e. uninfected, with single and double infections, either E. paraensei first (E + A) or A. cantonensis first (A + E) were followed; three times a week during four weeks the numbers of egg masses, eggs and hatched snails were counted. Histological analysis of the ovotestis and albumen gland was performed after four weeks. The number of egg masses/snail, eggs/snail and hatched snails showed significative differences comparing the control group to all infected groups, especially in group E + A, with the majority of values of parameters analyzed lower than 50% of those observed for control snails. In addition, the mortality was higher in the group E + A than to control group. The histological analysis showed that presence of both parasites in the reproductive organs was associated with tissue damages.

Comparative pattern of growth and development of Echinostoma paraensei (Digenea: Echinostomatidae) in hamster and Wistar rat using light and confocal laser scanning microscopy.

Echinostoma paraensei (Digenea: Echinostomatidae) lives in the duodenum and bile duct of rodents and is reported as a useful model for studies on the biology of flatworms. Here, we compared the growth and development of pre and post ovigerous worms collected 3, 7, 14 and 21 days post infection from experimentally infected hamster (permissive host) and Wistar rat (less permissive hosts). Linear measurements and ratios were examined by light (morphology and morphometry) and confocal laser scanning microscopy. At day 3, either worm from hamsters or rats were small with poorly developed gonads. At seven day, worms increased in size and morphometric differences between hosts are statistically significant after this time. In addition, adult worms (14 and 21 days of age) harvested from hamster showed developed gonads and vitelline glands laterally distributed on the body, whereas worms from rat showed atrophied reproductive system characterized by underdeveloped vitelline glands and stunted ovary. The worm rate recovery in rat decreased from 29.3% (day 7) to 20.6% (day 14) and 8% (day 21), whilst it remained around 37% in hamster. In conclusion, this is the first appointment demonstrating that low permissiveness influences the reproductive system of echinostome since the immature stages of development. The phenotypic analysis evidenced that hamster provides a more favorable microenvironment for gonads development than rat, confirming golden hamster as a permissive host, whereas Wistar rat is less permissive host.

Light and scanning electron microscopy study of in vitro effects of artesunate in newly excysted metacercariae of Echinostoma paraensei (Trematoda: Digenea).

Chemotherapy of food-borne trematodes relies on two drugs, praziquantel and tricabendazole, and there is growing interest in finding alternative therapies. Plant oil extracts have long been used in traditional Chinese medicine as sources of bioactive compounds with antiparasitic activity. Species of the genus Echinostoma are used as good models to test effective compounds against food-borne trematodes. This study evaluated the anthelmintic activity of crude artesunate extracts in vitro on newly excysted metacercariae of Echinostoma paraensei by light and scanning electron microscopy (SEM). The flukes were incubated with 1 µg/mL, 10 µg/mL, 25 µg/mL, 50 µg/mL and 100 µg/mL of artesunate for 4, 12, 24, 48 and 72 h. When the exposure time and concentration of artesunate increased, there were changes in motor activity, tegument damage and death. Blebs and swelling were the most common damages quantified on the tegument. The in vitro study reproduced results described for other immature flukes incubated with artemisinin derivatives. Excysted metacercariae of E. paraensei constitute a good model to study in vitro drug effects.

Effects of experimental Angiostrongylus cantonensis infection on the reproductive biology of Biomphalaria straminea and Biomphalaria tenagophila.

Eosinophilic meningoencephalitis is an endemic zoonosis in Southeast Asia and the Pacific Islands, but in recent years, new cases have been reported in various countries outside these regions, including Brazil, where it is considered an emerging disease. In this study, the effect of infection by the nematode Angiostrongylus cantonensis, one of the main etiologic agent of this disease, on the reproductive biology of the planorbid snails Biomphalaria straminea and B. tenagophila was investigated during the pre-patent period. Alterations in the reproductive biology of B. straminea and B. tenagophila were analyzed in laboratory-reared specimens infected by A. cantonensis during 21days; the number of eggs, number of egg masses, number of eggs/mass, number of eggs/snail, viable eggs/snail, survival and galactogen content in the albumen gland were measured. The results indicated the occurrence of initial compensation in reproductive effort in both snail species, but at different moments in the pre-patent period. More specifically, a reduction of 46.53% in the eggs/egg mass ratio in infected B. straminea was observed, a reflection of a 50% decline in the concentration of galactogen contained in the albumen gland. Changes in this parameter were also noted in B. tenagophila, but only at the end of the study period, with a reduction of 15.49%. Histological analyses indicate that changes observed can be explained by the tissue damages caused by the migration and development of the larvae. These results shed more light on the host-parasite relationship and indicate the importance of studying reproductive aspects for efforts to control infected snails. Considering that terrestrial snails can also transmit eosinophilic meningitis (in addition to aquatic mollusks), the data obtained expand knowledge of this host-parasite relationship and provide support for programs to control this zoonosis.

The influence of Angiostrongylus cantonensis (Nematoda, Metastrongylidae) infection on the aerobic metabolism of Biomphalaria straminea and Biomphalaria tenagophila (Mollusca, Gastropoda).

Angiostrongylus cantonensis is considered the main agent responsible for human eosinophilic meningoencephalitis. This parasite has low specificity for mollusk hosts and it can also use aquatic snails as auxiliary hosts. Studies based on the metabolic profile of Biomphalaria spp. infected by A. cantonensis have been conducted to observe parasite-host interactions. In the present study, the glucose content in the hemolymph and glycogen content in the digestive gland and cephalopedal mass of Biomphalaria tenagophila and Biomphalaria straminea experimentally infected by A. cantonensis were evaluated, along with the activity of LDH. The snails were dissected from 6 to 21days after infection to collect the hemolymph and separate the tissues. Decreases of 96% and 6.4% in the glucose content triggered a transition from aerobic to anaerobic metabolism in the two infected snail species, B. straminea and B. tenagophila, respectively. That finding was confirmed by high-performance liquid chromatography. These results indicate that when infected, these snails are able to change their metabolic profile, suggesting a strategy to maintain their homeostatic balance.

WITHDRAWN: Alterations in the mitochondrial physiology of Biomphalaria glabrata (Mollusca: Gastropoda) after experimental infection by Angiostrongylus cantonensis (Nematoda: Metastongylidae).

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In vivo and in vitro effects of the herbicide Roundup(®) on developmental stages of the trematode Echinostoma paraensei.

The exposure of wildlife and humans to toxic residues of Roundup(®) through agricultural practices or the food chain has been reported since the herbicide was found contaminating rivers. Glyphosate, N-(phosphonomethyl)glycine acid, is a nonselective post-emergent herbicide and is formulated as an isopropylamine salt with the surfactant taloamine polyethoxylate (POEA) representing the commercial formulation of Roundup(®). There is little knowledge about the effects of the herbicide on helminth parasites, particularly those whose life cycle is related to water bodies. Here we investigated the effects of the Roundup(®) on the food-borne trematode Echinostoma paraensei in experimental conditions using different developmental stages (eggs, miracidia, cercariae, metacercariae, newly excysted larvae (NEL), helminths at seven days and helminths at fourteen days). Three different herbicide concentrations were tested based on concentrations typically applied in the field: 225, 450 and 900 mg/L. Specimens were analyzed in vitro for hatching miracidia, mortality and excystment rate of metacercariae and in vivo for parasitic load and egg production. There was a significant difference in the hatching miracidia rate only for the newly embryonated eggs. The mortality of specimens and excystment rate of metacercariae were concentration-dependent. There was a significant difference in the miracidia mortality with respect to concentration until 56.3 mg/L. The same effect was observed for cercariae, and mortality was observed from 15 min onwards at concentrations of 225-900 mg/L. At low concentrations, mortality was detected after 30 min. The effects of the herbicide concentration on NEL and on helminths at seven and fourteen days showed a significant difference after 24 h. There was no significant difference in parasitic load and egg production after infection of rodents with exposed metacercariae. All developmental stages of the trematode E. paraensei were affected by Roundup(®) exposure under experimental conditions. These results suggest that dynamics of transmission of the trematode could be affected in the natural environments. The study also reinforces the usefulness of this trematode as a good model organism to test pesticides regarding human and environmental health.

Evaluation of the mitochondrial system in the gonad-digestive gland complex of Biomphalaria glabrata (Mollusca, Gastropoda) after infection by Echinostoma paraensei (Trematoda, Echinostomatidae).

The effect of infection by Echinostoma paraensei on the mitochondrial physiology of Biomphalaria glabrata was investigated after exposure to 50 miracidia. The snails were dissected one, two, three and four weeks after infection for collection and mechanical permeabilization of the gonad-digestive gland (DGG) complex. The results obtained indicate that prepatent infection by this echinostomatid fluke significantly suppresses the phosphorylation state (respiratory state 3) and basal oxygen consumption of B. glabrata, demonstrating that the infection reduces the ability of the intermediate host to carry out aerobic oxidative reactions. Additionally, relevant variations related to the uncoupled mitochondrial (state 3u) of B. glabrata infected by E. paraensei were observed. Four weeks after exposure, a significant reduction in mitochondrial oxygen consumption after addition of ADP (3.68±0.26pmol O2/mg proteins) was observed in the infected snails in comparison with the respective control group (5.14±0.25). In the uncoupled state, the infected snails consumed about 62% less oxygen than the infected snails (7.87±0.84pmol O2/mg proteins) in the same period. These results demonstrate a reduction in oxidative decarboxylation rate of the tricarboxylic acid cycle and faster anaerobic degradation of carbohydrates in the infected snails. The possible mechanisms that explain this new metabolic condition in the infected organisms are discussed.

Biochemical and histopathological alterations in Biomphalaria glabrata due to co-infection by Angiostrongylus cantonensis and Echinostoma paraensei.

The effect of concurrent infection by Echinostoma paraensei and Angiostrongylus cantonensis on the activity of the enzymes alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and the concentration of total proteins, uric acid and urea in the hemolymph of Biomphalaria glabrata were investigated. Additionally, histopathological studies were conducted to better understand the dynamics of ontogenic development of both helminths in the host and the possible biochemical effects. Co-infections by helminths and other parasites often occur due to the wide distribution of helminths and the chronic nature of the infection. The biochemical parameters were measured at the end of the seventh week after exposure. The co-infection resulted in a significant decrease in the total proteins concentration in the hemolymph of snails as well as an increase in the nitrogen excretion products, these results showed that the infection leads to exhaustion of free circulating and stored carbohydrates and the infected snails make use alternative substrates, such as free amino acids. So, the protein degradation to release free amino acids causes a decrease in the content of total proteins in the snail host and an amino acids deamination process, increasing the content of ammonium, which needs to be detoxified. This occurs by increasing the urea and uric acid contents. This observation is corroborated by the increase of ALT and AST activities, enzymes directly related to amino group from an amino acid to an α- ketoacid an important step to generate new carbon skeleton for glucose synthesis de novo, as well as new intermediates to the Krebs cycle. Additionally, reduction in the recovery of L3 from the co-infected group (A + E) was observed, since in this association the burden was higher than in the other. Histopathological results showed a change in the distribution of A. cantonensis in the presence of E. paraensei, indicating that the presence of this trematode impairs the dynamic transmission of A. cantonensis.

In vitro excystation of Echinostoma paraensei (Digenea: Echinostomatidae) metacercariae assessed by light microscopy, morphometry and confocal laser scanning microscopy.

Trypsin and bile salts have been identified as important triggers for excystation of Echinostoma metacercariae. Although excystation in trematodes is a well-known phenomenon, some morphological developmental changes remain to be elucidated. In order to gain further insight into the in vitro development of metacercariae, we assayed different cultivating conditions: 0.5% trypsin and 0.5% bile salts; 1% trypsin and 1% bile salts; 1% trypsin and 0.5% bile salts; 0.5% bile salts; or 0.5% trypsin. By means of light microscopy and confocal microscopy, we characterized each encysted, activated, breached and excysted stage based on the morphological features. However, breached and excysted stages were not revealed in both bile salts and trypsin-free medium. Excretory concretions (25 ± 3.9) were visualized within excretory tubules, close to the ventral sucker and genital anlage. The oral sucker armed with spines and digestive system was similar to those of adult worms. The reproductive system is composed of a genital anlage and the cirrus sac primordium. In short, trypsin and bile salts associated were fundamental for the in vitro metacercariae excystation of Echinostoma paraensei. This article presents the first detailed information of all stages of metacercariae excystation obtained through light and confocal microscopy.

Aerobic to anaerobic transition in Biomphalaria glabrata (Say, 1818) infected with different miracidial doses of Echinostoma paraensei (Lie and Basch, 1967) by high-performance liquid chromatography.

The glucose content in the hemolymph and glycogen content in the digestive gland-gonad complex (DGG) and cephalopedal mass of Biomphalaria glabrata exposed to different parasite doses (5 and 50 miracidia) of Echinostoma paraensei as well as the activity of lactate dehydrogenase were evaluated. HPLC (high-performance liquid chromatography) analyses were also performed to determine the concentrations of four organic acids (oxalic, succinic, pyruvic and lactic) present in the hemolymph of infected and uninfected snails, to better understand the effect of infection on the host's energetic/oxidative metabolism. The snails were dissected 1-4 weeks after infection to collect the hemolymph and separate the tissues. There was alteration in the glycemia of the snails at both parasite doses, with a significant increase of glycemia from of the third week after infection in comparison to the control group. Changes were also observed in the lactate dehydrogenase activity, with increased activity as the infection progressed. In parallel, there was a decrease in the glycogen content in the storage tissues, with a markedly greater reduction in the digestive gland-gonad complex (larval development site) in comparison with the cephalopedal mass. Additionally, the infection by both miracidial doses resulted in an increase of oxalic and lactic acid levels, as well as in a decline of piruvic and succinic acid levels in B. glabrata, thus explaining the reduction of the oxidative decarboxylation rate in the tricarboxylic acid cycle and acceleration of the anaerobic degradation of carbohydrates in the snails, through lactic fermentation, which is essential to ensure energy supply and success of the infection.

Workshop on research priorities for management and treatment of angiostrongyliasis(1).

In a concluding session of the workshop, the participants developed a list of 115 research and outreach needs, outlining the top 5-7 needs in each of 8 areas (Table). For complete information, including presenter details and abstracts, visit the workshop website at www.hawaii.edu/cowielab/Angio%20website%20home.htm.

Genetic and morphological characterisation of a new species of the genus Hysterothylacium (Nematoda) from Paralichthys isosceles Jordan, 1890 (Pisces: Teleostei) of the Neotropical Region, state of Rio de Janeiro, Brazil.

Taking into account the difficulties of taxonomic identification of larval anisakid nematodes based on morphological characters, genetic analyses were performed, together with those usually applied, in order to identify anisakid larvae found in the flounder Paralichthys isosceles from the littoral of the state of Rio de Janeiro, Brazil. The analysis of 1,820 larvae revealed a new species, similar to Hysterothylacium MD, Hysterothylacium 2, Hysterothylacium KB and Hysterothylacium sp regarding the absence of the larval tooth, an excretory pore situated below the nerve ring level, and slender lateral alae. Moreover, the new species differs from Hysterothylacium fortalezae and Hysterothylacium reliquens with regard to the number and size of spines present on the tail end and from Hysterothylacium patagonicus by the absence of interlabia. The maximum parsimony and neighbour joining tree topologies based on the 18S ribosomal DNA gene, complete internal transcribed spacer region and cytochrome oxidase 2 (COII) gene demonstrated that the Brazilian larvae belong to Raphidascarididae and represent a unique genetic entity, confirmed as a new Hysterothylacium species. Furthermore, the new species presents COII genetic signatures and shares polymorphisms with Raphidascarididae members. This is the first description of a new anisakid species from Brazil through the integration of morphological and molecular taxonomy data.

Effect of praziquantel on adult Echinostoma paraensei worms in experimentally infected mice.

Echinostomiasis is a food-borne intestinal, snail-mediated parasitosis caused principally by ingestion of snails infected with digenean trematodes of the Echinostoma genus. The treatment and control of trematodiasis is usually done by administration of praziquantel (PZQ). In this study, we evaluated the effect on Echinostoma paraensei of different doses of praziquantel through analysis of morphological parameters using light microscopy, scanning electron microscopy, and confocal scanning laser microscopy along with parasitological data. We used 30 female mice aged 4 weeks. Each animal was given 40 metacercarie of E. paraensei by gavage. The animals were divided into five groups, each group containing six animals, where one group was utilized as untreated control. Two weeks after infection, the mice were given praziquantel by gavage at total dosages of 12.5, 25, 50 or 100 mg/kg by body weight. Two days after treatment, the mice were euthanized in a CO(2) chamber for recovery of helminths in the small intestine. The doses of 50 and 100 mg/kg of praziquantel eliminated all the worms. There were significant differences (p<0.05) between all the treated groups when compared to the control group. The body morphology showed contraction with vacuolization of the parenchyma, and the spine of the peristomic collar was not evident by light microscopy. The scanning electron microscopy revealed that the other doses caused retraction of spines of the peristomic collar and also the tegument spines at the body edge, as well as the development of vesicles and peeling; all these alterations were more evident at the dose of 25 mg/kg. In turn, the confocal scanning laser microscopy revealed vacuolization and disorganization of spines and vitelline glands. E. paraensei responds differently to experimental treatment with praziquantel according to the doses utilized causing morphological alteration and even worm elimination.

New Pudicinae (Nematoda, Heligmonellidae) parasite of Trichomys fosteri (Rodentia: Echimyidae) from Pantanal, Brazil.

Pudica trichomysae n. sp. (Trichostrongylina, Heligmosomoidea, Helligmonellidae, Pudicinae) from the small intestine of Trichomys fosteri (Rodentia: Echimydae) from the Pantanal of Mato Grosso do Sul is described by light and scanning electron microscopy. Pudica trichomysae n. sp. is characterized by caudal bursa type 2-2-1, proportion of spicules length in relation to body length (SpL/BL) of 17 to 18% left and right, respectively. Dorsal ray divided at about the distal third into two branches, each branch divided into two long sub-branches, rays 9 and 10. Furthermore, this study shows for the first time details of the anterior region demonstrating papillae organization, amphids, derides, and opening of the excretory pore. The posterior end of the caudal bursa showed detail of the genital cone, papillae, and ray 1, and in the female, it showed interrupted ridges between the vulva and anus. In conclusion, the present species is the seventeenth described in the genus in South America and the first in Pantanal.

A new species of Pterygodermatites (Spirurida: Rictulariidae) in Marmosa constantiae Thomas, 1904 from an ecotone area of the biomes Cerrado/Amazon in the Mato Grosso State, Brazil.

A new species of nematode, Pterygodermatites (Paucipectines) sinopiensis n. sp. is described based on specimens recovered from the intestine of the white-bellied woolly mice opossum, Marmosa constantiae, trapped in the municipality of Sinop, Mato Grosso state, Brazil. The genus Pterygodermatites has 21 species described in mammals worldwide, and to date, only two species have been described for marsupials in Brazil. The new species is characterized by the presence of 23 small denticles and by the presence of 38-40 and 65 pairs of the cuticular processes in male and female species, respectively. Additionally, male species possess three ventral precloacal fans, and in female species, the cuticular processes are divided into 41 pairs of comb-like and 24 pairs of spine-like processes; the vulva opens approximately in pair 41. This study describes the parasite species fifth of marsupials in the Neotropical region.

MORPHOLOGICAL AND GENETIC CHARACTERIZATION OF DIDELPHONEMA LONGISPICULATA (NEMATODA: SPIRUROIDEA) IN THE BLACK-EARED OPOSSUM DIDELPHIS MARSUPIALIS.

Didelphonema longispiculata (Hill, 1939), a gastric nematode parasite of the black-eared opossum, Didelphis marsupialis Linnaeus, 1758, collected from 2 municipalities of Mato Grosso state, Brazil, in the ecotone region of the Amazon and Cerrado biomes was analyzed with integrative taxonomy using light and scanning electron microscopy (SEM) for morphological studies and sequencing of the 18S small subunit ribosomal RNA for phylogenetic inference through maximum likelihood and Bayesian phylogenetic inference. Here details of the helminth surface, oral aperture with octagonal border, pseudo- and inter-labia, amphids, external cephalic papillae, 2 dorsal and ventral internal plates distally indented, and stoma with strongly chitinized wall are presented. Caudal male papillae, spicules, female vulva, anus, and caudal tip were detailed using SEM. Morphological characteristics and phylogenetic data corroborated the taxonomic placement of the genus Didelphonema within the subfamily Ascaropsinae.

Development of the reproductive system of Echinostoma paraensei in Mesocricetus auratus analyzed by light and confocal scanning laser microscopy.

This study was performed to gain insight into the maturation of the reproductive system of Echinostoma paraensei worms grown in an early infection of Mesocricetus auratus. Hamsters were infected with 100 metacercariae and necropsied on days 3, 5, 7, 10 and 14 post infection (dpi). Recovered flukes stained with hydrochloric carmine were preserved as whole mounts and analyzed by light and confocal scanning laser microscopy. The average worm recovery was 43.7 per host. Images of the male and female reproductive systems were taken. The ovary and anterior and posterior testis were evidenced on day 3, while the ootype and cirrus sac were present on day 5. Confocal imaging showed primordium testis and ovary as a cluster of primordial cells from day 3 onward. The testes, ovary, cirrus sac and uterus organs were already present during the first week of life. The two testes were seen as individual structures on 7 dpi while the cirrus sac and vitelline glands were in development. The ovary was connected to the uterus while the ootype was adjacent to it. Both testes were larger than the ovary, showing cells at different stages of development, but with few bundles of functional spermatozoa in 10 day-old worms. On day 14, eggs and spermatozoa were seen in the uterus and seminal vesicle, respectively, while oocytes appeared in the ootype as fertilized eggs. We conclude that the reproductive system of E. paraensei was functional on 14 dpi in the hamsters.