Smp38 MAP Kinase Regulation in Schistosoma mansoni: Roles in Survival, Oviposition, and Protection Against Oxidative Stress.

Eukaryotic protein kinases (ePKs) are good medical targets for drug development in different biological systems. ePKs participate in many cellular processes, including the p38 MAPK regulation of homeostasis upon oxidative stress. We propose to assess the role of Smp38 MAPK signaling pathway in Schistosoma mansoni development and protection against oxidative stress, parasite survival, and also to elucidate which target genes have their expression regulated by Smp38 MAPK. After a significant reduction of up to 84% in the transcription level by Smp38 MAPK gene knockdown, no visible phenotypic changes were reported in schistosomula in culture. The development of adult worms was tested in vivo in mice infected with the Smp38 knocked-down schistosomula. It was observed that Smp38 MAPK has an essential role in the transformation and survival of the parasites as a low number of adult worms was recovered. Smp38 knockdown also resulted in decreased egg production, damaged adult worm tegument, and underdeveloped ovaries in females. Furthermore, only ~13% of the eggs produced developed into mature eggs. Our results suggest that inhibition of the Smp38 MAPK activity interfere in parasites protection against reactive oxygen species. Smp38 knockdown in adult worms resulted in 80% reduction in transcription levels on the 10th day, with consequent reduction of 94.4% in oviposition in vitro. In order to search for Smp38 MAPK pathway regulated genes, we used an RNASeq approach and identified 1,154 DEGs in Smp38 knockdown schistosomula. A substantial proportion of DEGs encode proteins with unknown function. The results indicate that Smp38 regulates essential signaling pathways for the establishment of parasite homeostasis, including genes related to antioxidant defense, structural composition of ribosomes, spliceosomes, cytoskeleton, as well as, purine and pyrimidine metabolism pathways. Our data show that the Smp38 MAPK signaling pathway is a critical route for parasite development and may present attractive therapeutic targets for the treatment and control of schistosomiasis.

Phenotypic plasticity of male Schistosoma mansoni from the peritoneal cavity and hepatic portal system of laboratory mice and hamsters.

Morphometric analysis of Schistosoma mansoni male worms obtained from AKR/J and Swiss mice was carried out. Rodents infected by the intraperitoneal route with 80 cercariae of the schistosome (LE strain) were killed by cervical dislocation at 45 and 60 days post-infection and both peritoneal lavage and perfusion of the portal system were performed for the recovery of adult worms. Characteristics including total body length, the distance between oral and ventral suckers, extension of testicular mass and the number of testes were considered in the morphological analysis. Changes that occurred in S. mansoni recovered from the peritoneal cavity or from the portal system of AKR/J and Swiss mice included total body length and reproductive characteristics. Significant morphometric alterations were also observed when worms recovered from the portal system of both strains of mice were compared with the schistosomes obtained from hamsters (Mesocricetus auratus), the vertebrate host in which the LE strain had been adapted and maintained by successive passages for more than four decades. The present results reinforce the idea that S. mansoni has high plastic potential and adaptive capacity.

Morphological and morphometric study of cercariae and adult worms of Schistosoma mansoni (SLM strain) isolated from infected mice.

In northeastern Brazil, the schistosomiasis is historically endemic and considered as a public health problem. The Schistosoma mansoni São Lourenço da Mata (SLM-PE, Brazil) strain was used in several paper already published; however, morphological and morphometric studies about this strain was never done. In this work, scanning electron microscopy (SEM) was used in morphological and morphometric analysis of cercariae and adult worms. Cercariae were obtained from Biomphalaria glabrata snails and adult worms from mice, both infected by the S. mansoni SLM strain, fixed and prepared for SEM. The results showed that cercariae of S. mansoni measures 254.9 µm of length. The bodies are covered by spines, with a ventral sucker, an oral sucker with sensory receivers, and a pair of penetration glands in the head. The area of tail and body and the distance between suckers were 3,011.77, 1,530.32, and 42.9 µm, respectively. Adult worms of S. mansoni were divided into three main regions: the anterior, medial, and posterior, besides the gynecophoral canal in males. The measure of adult worms of S. mansoni was 4 mm males and 5 mm females. The anterior region length of the male was 470 µm and of the female 271 µm. All the parameters were assayed in ten samples. The morphometric values found in the SLM strain were smaller than other S. mansoni strains described in the literature as well as other helminths. This is the first morphological and morphometric study with the SLM strain of S. mansoni being extremely important for improving control strategies and life quality of the local population.

Anthelmintic activity of carvacryl acetate against Schistosoma mansoni.

Blood flukes of the genus Schistosoma are the causative agents of human schistosomiasis, a debilitating disease that afflicts over 200 million people worldwide. Praziquantel is the drug of choice but concerns over praziquantel resistance have renewed interest in the search for alternative drug therapies. Carvacrol, a naturally occurring monoterpene phenol and food additive, has been shown high medicinal importance, including antimicrobials activities. The aim of this study was to evaluate in vitro effect of carvacryl acetate, a derivative of carvacrol, on Schistosoma mansoni adult worms. We demonstrated that carvacryl acetate at 6.25 µg/mL has antischistosomal activity, affecting parasite motility and viability. Additionally, confocal laser scanning microscopy pictures revealed morphological alterations on the tegumental surface of worms, where some tubercles appeared to be swollen with numerous small blebs emerging from the tegument around the tubercles. Furthermore, experiments performed using carvacryl acetate at sub-lethal concentrations (ranging from 1.562 to 6.25 µg/mL) showed an inhibitory effect on the daily egg output of paired adult worms. Thus, carvacryl acetate is toxic at high doses, while at sub-lethal doses, it significantly interferes with the reproductive fitness of S. mansoni adult worms. Due to its safety and wide use in the industry, carvacryl acetate is a promising natural product-derived compound and it may represent a step forward in the search for novel anthelmintic agents, at a time when there is an urgent need for novel drugs.

Parasitological and morphological study of Schistosoma mansoni and diabetes mellitus in mice.

Schistosomes are blood-dwelling flukes which are highly dependent on the host metabolism. The aim of this study was to investigate possible relationship between streptozotocin-induced diabetes and the outcome of acute murine schistosomiasis mansoni. Male and female SW mice were treated by a single intraperitoneally injected dose of streptozotocin (180 mg/kg). Seven days after induction, both control and diabetic animals were infected with 70 Schistosoma mansoni cercariae (BH strain). Diabetics and their controls were weighed 45 days after birth and for the last time prior to killing. Susceptibility to infection was evaluated twice a week by quantifying fecal egg excretion 7-9 weeks post-infection by the Kato-Katz' thick smear method. Mice were euthanized the day after the last fecal examination was performed. Adult worms were recovered from the portal system and mesenteric veins, whereas liver and intestine were removed for enumeration of egg load. No differences in worm length or in measurements of the reproductive organs, tegument, and suckers were detected. Also oviposition was unaffected as the total number of eggs per female worm from the liver, the small and the large intestine was the same in both groups. An oogram evaluation revealed a lower percentage of mature (23.0% vs. 40.7%) and a higher percentage of immature (69.1% vs. 51.7%) eggs in the small intestine of the diabetic mice. We suggest that principally a hampered egg passage through the intestine tissue caused this reduction and that probably both the eggs and the impaired host response play a role.

Schistosoma mansoni Molecular characterization of Alkaline Phosphatase and expression patterns across life cycle stages

Here we describe the cloning and characterization of the Schistosoma mansoni Alkaline Phosphatase(SmAP), previously identified in the tegument of adult worms. SmAP encodes a complete sequence composedof 536 amino acids containing an N-terminal signal peptide, five N-glycosylation sites, and a GPIanchor signal, similar to that described for mammalian orthologs. Real-time RT-PCR and Western blotexperiments suggest a rapid translation as soon as cercariae are transformed into schistosomula. Immunolocalizationanalysis shows that the protein is widely distributed in the worm tissues, with increased concentration in the vitelline glands of female parasites. Furthermore, the surface localization of thisenzyme was quantitatively supported by its enzymatic activity in live ex vivo or cultured parasites throughout the life cycle stages. The fact that cercariae accumulate large amounts of SmAP mRNA, which rapidly translates into protein upon schistosomula transformation, indicates it may have an important role in host invasion.

Do schistosomes grow old? A confocal laser scanning microscopy study.

Infected hosts progressively decrease egg output during the chronic phase of Schistosoma mansoni infection. Ageing may be a factor that results in a progressive decrease in the ability to reproduce. This study was performed to gain insight into the effects of ageing on adult schistosomes, using confocal laser scanning microscopy. Adult worms were recovered from experimentally infected Nectomys squamipes (water rat). Viable eggs were voided in the faeces for 65 weeks and, thereafter, only unviable eggs were eliminated. The rat died after 70 weeks in captivity. Recovered worms (six males and one female) were prepared for confocal microscopy and images were obtained with an LSM 510-ZETA laser confocal microscope. The overall morphology of the adult worms (suckers and tegument) and reproductive organs were the focus of this work. The suckers, ovary, vitellaria and oocytes appeared to be apparently normal. The ootype was formed by flattened cells and unclear nuclei, suggesting that the mechanism for eggshell production by the ootype during organogenesis might have been impaired. Testicular lobes showed empty areas around the germinative cells. Male adult worms showed flaccid musculature of the dorsal surface. Taken together, the present results provided several indications of infertility in older male and female schistosomes.

Development of Schistosoma mansoni in the laboratory rat analyzed by light and confocal laser scanning microscopy.

The kinetic of maturation (schistogram) of Schistosoma mansoni worms grown in laboratory rats was studied by light and confocal laser scanning microscopy. Infected rats with the BH strain were weekly euthanized 3-9weeks pi. Recovered flukes stained with hydrochloric carmine were preserved as whole-mounts and analyzed by confocal and brightfield microscopy. Worms displayed varying degrees of maturation of the reproductive system at weeks 3-6. Male worms showed complete maturation of the reproductive system at week 6, while female worms completed their maturation at week 7. Males presented few tubercles in tegument in all weeks. Despite the presence of a developing embryo within the ootype, no uterine egg was found. The schistogram in rats follows a pattern similar to that observed in mice hosts.

Descrição do gene do receptor de TNF-α Schistosoma mansoni e efeito do TNF-α humano na expressão gênica do parasita / Description of the TNF-α receptor gene in Schistosoma mansoni and the effect of human TNF-α on the parasite's gene expression

O parasita Schistosoma mansoni é um dos principais causadores da esquistossomose, doença que acomete 200 milhões de indivíduos no mundo. O parasita possui um complexo ciclo de vida composto de seis estágios evolutivos em dois hospedeiros. S. mansoni possui um sofisticado sistema de interação com seus hospedeiros de modo a escapar da resposta imune e interagir com moléculas produzidas por eles. Alguns trabalhos na literatura descreveram o efeito do TNF-α humano sobre o processo de ovoposição do parasita adulto. Nosso trabalho teve por objetivo analisar o perfil de expressão gênica do S. mansoni ao longo de seus estágios de desenvolvimento, avaliar o efeito do TNF-α humano sobre o perfil de expressão gênica do parasita em dois estágios de desenvolvimento e descrever o gene homólogo ao receptor de TNF-α humano em S. mansoni. Para isso, duas plataformas de microarrays distintas foram utilizadas: uma composta por 4000 sondas de cDNA dupla fita produzida pelo nosso grupo de pesquisa e a outra, composta por 44000 sondas de oligonucleotídeos desenhadas pelo nosso grupo e produzida pela empresa Agilent Technologies. Com estas plataformas foi detectada a expressão de 5798 genes em vermes adultos, sendo que 156 genes apresentavam a transcrição nas fitas senso e anti-senso; 6 destes tiveram confirmadas a transcrição nas duas fitas por transcrição reversa fita específica seguida de PCR em Tempo Real. Adicionalmente foram identificados 229 genes diferencialmente expressos entre vermes adultos machos e fêmeas. A análise de expressão gênica entre 5 estágios de desenvolvimento do parasita mostrou dois tipos de conjuntos de dados: (i) 1423 genes diferencialmente expressos entre dois estágios subseqüentes de desenvolvimento e (ii) 342 genes com expressão enriquecida em um estágio exclusivamente. 68 destes são transcritos intrônicos que não possuem potencial codificador de proteinas. Um gene ortólogo ao receptor de TNF-α humano (SmTNFR) foi clonado e...

The parasite Schistosoma mansoni is one of major causative agents of schistosomiasis, a disease which affects 200 million people in the world. The parasite has a complex life cycle with six developmental stages in two hosts. S. mansoni has a sofisticated system of interaction with the hosts, permitting it to escape the immune response and to interact with molecules produced by the hosts. The effect of human TNF-α on adult parasite egg-laying has been described in the literature. The present work intended to analyse the gene expression profile of S. mansoni among its developmental stages, to evaluate the effect of human TNF-α on gene expression profile in two different developmental stages and to describe a homologous gene to human TNF-α receptor in S. mansoni. Two microarrays platfoms were used: one comprised by 4000 cDNA probes and printed by our research group and another, comprised by 44000 oligonucleotide probes designed by our group and printed by Agilent Technologies Company. With these platforms, we detected the expression of 5798 genes in adult worms, of which 156 showed transcription in sense and anti-sense strands; 6 of them had their expression levels confirmed by strand specific Real Time PCR. 229 genes were identified as differentially expressed between male and female adult worms. Gene expression analysis among 5 parasite developmental stages identified two data sets: (i) 1423 differentially expressed genes between two subsequent developmental stages and (ii) 342 expressed genes enriched in one exclusive stage. 68 of them are intronic transcripts with no protein-coding potential. An ortologue to human TNF-α receptor (SmTNFR) was cloned and sequenced. SmTNFR transcritpt has 1967bp and encodes a 599-amino acid protein. Other 9 genes encoding conserved elements of the TNF-α signaling pathway were identified among the public S. mansoni ESTs dataset, thus revealing a complete TNF-α signaling pathway in the parasite...

Extracellular lipid droplets promote hemozoin crystallization in the gut of the blood fluke Schistosoma mansoni.

Hemozoin (Hz) is a heme crystal produced upon hemoglobin digestion as the main mechanism of heme disposal in several hematophagous organisms. Here, we show that, in the helminth Schistosoma mansoni, Hz formation occurs in extracellular lipid droplets (LDs). Transmission electron microscopy of adult worms revealed the presence of numerous electron-lucent round structures similar to LDs in gut lumen, where multicrystalline Hz assemblies were found associated to their surfaces. Female regurgitates promoted Hz formation in vitro in reactions partially inhibited by boiling. Fractionation of regurgitates showed that Hz crystallization activity was essentially concentrated on lower density fractions, which have small amounts of pre-formed Hz crystals, suggesting that hydrophilic-hydrophobic interfaces, and not Hz itself, play a key catalytic role in Hz formation in S. mansoni. Thus, these data demonstrate that LDs present in the gut lumen of S. mansoni support Hz formation possibly by allowing association of heme to the lipid-water interface of these structures.

Hermaphrodites and supernumerary testicular lobes in Schistosoma mansoni (Trematoda: Schistosomatidae) analyzed by brightfield and confocal microscopy.

Schistosoma mansoni belongs to the dioecious Schistosomatidae. The occasional observation of males with rudimentary female characteristics is thought to attest the hermaphroditic roots of this parasite. Supernumerary testicular lobes also are recurrently seen in this helminth, but their morphology and origin are elusive. Here, we investigated the morphology of the supernumerary lobes from 15 S. mansoni males and similar structures of 2 females in whole mounts by brightfield and confocal laser scanning microscopy. The follicles in the females were not regarded as testicular lobes, but 1 male had a rudimentary ovary with a residual oviduct posterior to the normal set of testicular lobes and was considered hermaphroditic. In addition, 6 other males contained supernumerary lobes that enclosed fully matured oocytes in variable amounts. In the remaining 8 males, no female characteristics could be seen, and the lobes showed morphologically a more testicular aspect, although this morphology was only confirmed in 1 male where spermatozoids were detected. The process underlying the expression of supernumerary testes or lobes with oocytes is similar and is, at least in part, induced by the host.

Phenotypic plasticity in the cercarial indices of human and rodent samples of Schistosoma mansoni (Digenea: Schistosomatidae).

In the epidemiology of schistosomiasis, the cercarial index (CI) has been used for several purposes, including the identification of the species of vertebrate host involved in transmission in a particular setting. The CI of samples of Schistosoma mansoni of human (BH) and rodent (CMO) origin have now been compared, in an attempt to identify morphological markers on cercariae that would indicate whether the eggs from which the cercariae have developed were produced in humans or in rodents. After silver-nitrate impregnation of the cercariae, the CI (and their reaction norms) for the antero-dorsal (AD), antero-lateral (AL) and tail-stem (U) sensory receptors were investigated. The data analysis was based on linear-regression methods, Levene's test for the equality of variances, pooled-variance independent t-tests, and discriminant analysis. The results indicated that CI varied with vertebrate-host origin, that the AD reaction norms were more consistent and plastic than the other reaction norms investigated, that there appeared to be no simple allometric relationship between any of the CI investigated and cercarial body length, and that the BH sample gave larger standard deviations and a higher incidence of mis-classification than the CMO, indicating some developmental instability in the sample of human origin. The relative instability of the BH sample was, however, possibly the result of the experimental procedures. As the parasites used to produce the BH sample had been isolated from humans fairly recently and then routinely maintained in mice, host-induced selection could have affected the sample, altering its CI and their reaction norms. The results of the discriminant analysis indicated that, of the indices investigated, AD was the best morphological marker for distinguishing human and rodent samples of S. mansoni.

Do mice genetically selected for resistance to oral tolerance provide selective advantage for Schistosoma mansoni infection?

A highly evolved relationship exists between the parasitic flatworm Schistosoma mansoni and its vertebrate hosts that include the use of host immune signals by parasites. The S. mansoni infection was studied in two strains of mice genetically selected, over 18 generations of assortative mating, for extreme phenotypes of susceptibility (TS) and resistance (TR) to immunological tolerance. The objective was to observe whether the different host genetic backgrounds affected the outcome of experimental schistosomiasis. Fecal egg excretion, tissue egg count, worm recovery, and adult worm morphology and morphometry were monitored throughout the period of infection. TR mice presented total fecal egg excretion and thickness of tegument in adult male worms significantly higher than TS mice. Therefore, the comparative analysis of mice with extreme phenotypes of immunological response turns out to be useful in host-parasite relationship studies. Our results suggest that the TR mouse immunological profile provides a more favorable environment for the development of S. mansoni.

A new description of the reproductive system of Schistosoma mansoni (Trematoda: Schistosomatidae) analyzed by confocal laser scanning microscopy.

Classical schemes of the adult Schistosoma mansoni reproductive system have been described. In our study, whole adult worms derived from unisexual or mixed infections and stained with carmine chlorine were virtually and tomographically analyzed under confocal laser scanning microscopy. We found that: (1) there were morphological differences in the ovary, vitteline glands and testicular lobes between specimens derived from unisexual or mixed infections; (2) there was always a single lobed ovary (three or four lobes), presenting differentiation from the anterior to the posterior lobes, where the most mature oocytes were located; (3) the proximal segment of oviduct was connected to an ampullary dilatation, full of tailed spermatozoa, characterizing a seminal receptacle; (4) there was no long vitelline duct, but a short one that begins at the end of the proximal region of the vitelline gland; (5) long cells of Mehlis' gland placed radially around the ootype were not observed. Otherwise, the ootype was only lined by thick cuboidal epithelial cells with plaited bases and nuclei with flabby chromatin, making a clear distinction from the uterine epithelium. This morphological feature suggests that each cell represents a gland. (6) In coupled males, the specimens located inside the gynaecophoric canal had smaller testicular lobes, suckers, and body length and width when compared to their partners. Our results show that the reproductive system does not follow a unique pattern within flatworms. Due to its better resolution, confocal laser scanning microscopy, using a reflected mode with tomographic sections, allows new interpretations, modifying the adopted and current descriptions of the internal morphological structures of S. mansoni adult worms.

[Parasitological and morphological characteristics of Brazilian strains of Schistosoma mansoni in Mus musculus]. / Características biológicas e morfológicas de cepas brasileiras de Schistosoma mansoni em Mus musculus.

Aiming to determine parasitological and morphological differences between Brazilian strains (CMO, CM and BE) of Schistosoma mansoni the following parameters were studied: prepatent period, kinetics of egg releasing in feces, intestinal oogram, infectivity and the morphology of adult worms. No statistical difference (p>0.05) was found regarding the mean prepatent period (44 and 46 days) and infectivity (26% and 29%). All strains showed a higher concentration of trapped eggs in the small intestine than large intestine (p<0.05). All characteristics of the reproductive system and suckers of male worms presented significant differences (p<0.05). CMO strain was smaller in all measurements. Females showed significant differences (p<0.05) in the larger diameter of the eggs, in the area and perimeter of the spine egg and in the oral sucker area. We conclude that differences in strains can be expressed even when the strains have been cycled for several generations under laboratory conditions.

Morphometric study of Schistosoma mansoni adult worms recovered from Undernourished infected mice.

Some unfavourable effects of malnutrition of the host on Schistosoma mansoni worm biology and structure have been reported based upon brigthfield microscopy. This paper aims to study by morphometric techniques, some morphological parameters in male and female adult worms recovered from undernourished albino mice in comparison with parasites recovered from well-fed infected mice. Undernourished animals were fed a multideficient and essentially low protein diet (RBD diet) and compared to well-fed control mice fed with the commercial diet NUVILAB. Seventy-five days post-infection with 80 cercarie (BL strain) animals were sacrificed. All adult worms were fixed in 10% formalin and stained with carmine chloride. One hundred male and 60 female specimens from each group (undernourished and control) were examined using an image system analysis Leica Quantimet 500C and the Sigma Scan Measurement System. The following morphometrical parameters were studied: body length and width, oral and ventral suckers, number and area of testicular lobes, length and width of ovary and uterine egg. For statistical analysis, the Student's t test for unpaired samples was applied. Significant differences (p < 0.05) were detected in body length and width, in parameters of suckers, uterine egg width, ovary length and area of testicular lobes, with lower values for specimens from undernourished mice. The nutritional status of the host has negative influence on S. mansoni adult worms, probably through unavailability of essential nutrients to the parasites.

Morphological changes in the reproductive organs of male and female Schistosoma mansoni worms caused by streptozotocin, a drug used to induce diabetes mellitus.

Host metabolic changes have been observed to affect Schistosoma mansoni egg production, worm survival and morphology. We examined worms recovered from streptozotocin-induced diabetic mice by morphometric and morphological analysis through brightfield and confocal laser scanning microscopy. Tegument thickness was slightly smaller and changes in the reproductive organs were observed in 23-30% and consecutively 88-100% of the worms. The testicular lobes had a large diminution of cells in one or more of the lobes, which was associated with a lack of spermatozoids in the seminal vesicle. Ovaries were atrophied, manifested by a complete or large reduction in oocytes but other parts of the reproductive system like the vitelline glands were mainly unaffected. Streptozotocin (STZ) instead of hyperglycaemia caused the degeneration since worms from mice injected with a non-diabetogenic dose, or with nicotinamide to prevent diabetes showed the same alterations. The drug did not affect worm survival or pairing. We conclude that STZ, an alkylating agent that provokes chromosome and DNA damage, changes the morphology of ovaries and testicular lobes in S. mansoni worms in vivo. This is the first report of STZ action in helminths and we suggest that STZ affects oogenesis and spermatogenesis and might cause sterilization of schistosomes.

[Characterization of Schistosoma mansoni strains by morphometry of adult worms derived from single-sex infection]. / Caracterização de cepas de Schistosoma mansoni por morfometria de vermes adultos provenientes de infecção unissexual.

Mice were infected with only one sex cercaria derived from sympatric strains of Schistosoma mansoni. Adult worms presented significative differences (p < 0.05) regarding suckers, testicular lobes, ovary and thickness of the tegument. Data show that morphometric study of unisexual infection worms can be also used for characterization of Schistosoma mansoni strains.

Phenotypic characterization of Schistosoma mansoni adult worms recovered from undernourished mice: a morphometric study focusing on the reproductive system.

A morphometric study focusing on some features of the reproductive system of Schistosoma mansoni adult worms was performed, aiming to complete previously reported data concerning the effects of undernourishment of the host on the parasites. Male worms were significantly affected (p<0.05) regarding the testicular lobes.

[The size polymorphism of the cercariae of a Venezuelan strain of Schistosoma mansoni]. / Polimorfismo de tallas en las cercarias de una cepa venezolana de Schistosoma mansoni.

By mean of the silver impregnation of a lot of 47 cercariae belonging to C5 venezuelan Schistosoma mansoni strain, a polymorphism of size was evidenced. The morph was conformed by the greatest cercariae (between 152.9 microns to 218.4 microns). This group was ever present in the population of cercariae in less percentage (29.8%). The length of the normal cercariae was comprised between 119 microns to 152 microns. This group was the dominant in the population: 70.2%. The comparison between the length of both lots of cercariae resulted with significative statistical differences (t = 5.5; P < 0.001). For the total studied cercariae the average of the length was X = 143.9 microns and for the wide X = 68.6 microns. It was found a positive significative correlation between the wide and the length of the cercariae r = 0.83 (P < 0.001).