Susceptibility of embryos of Biomphalaria tenagophila (Mollusca: Gastropoda) to infection by Pochonia chlamydosporia (Ascomycota: Sordariomycetes).

Schistosoma mansoni is a heteroxenous parasite, meaning that during its life cycle needs the participation of obligatory intermediate and definitive hosts. The larval development occurs in aquatic molluscs belonging to the Biomphalaria genus, leading to the formation of cercariae, which emerge to infect the final vertebrate host. For this reason, studies for control of the diseases caused by digenetic trematodes often focus on combating the snail hosts. Thus, the objective of this study was to evaluate the susceptibility of Biomphalaria tenagophila embryos to the fungus Pochonia chlamydosporia (isolate Pc-10). The entire experiment was conducted in duplicate, with five replicates for each repetition (five egg masses/replicate), utilizing a total of 100 egg masses, with 20-30 eggs/egg mass. At the end of 15 days, the egg masses were evaluated under a stereomicroscope to analyze the hatching of B. tenagophila embryos in both experimental groups. After days of interaction, the exposure to the fungal hyphae bodies significantly impaired the viability of the B. tenagophila eggs, inhibiting the embryogenesis process by 83.7% in relation to the control group. Transmission and scanning electron microscopic images revealed relevant structural alterations in the egg masses exposed to the hyphae action of the fungus, interfering in the development and hatching of the young snails under analysis. These results indicate the susceptibility of B. tenagophila embryos to the fungus P. chlamydosporia (isolate Pc-10) and suggest the potential of Pc-10 to be used in the control of intermediate host, for its ovicidal capacity and for being an ecologically viable option, but in vivo experiments become necessary.

Physiological alterations in Pseudosuccinea columella (Mollusca: Gastropoda) after infection by Heterorhabditis baujardi LPP7 (Rhabditida: Heterorhabditidae).

The snail Pseudosuccinea columella participates in the distribution of Fasciola hepatica in the environment by acting as its intermediate host. Therefore, the control of this lymnaeid is one of the ways to prevent hepatic fascioliasis. The objective of this study was to evaluate the susceptibility of P. columella to infective juveniles (IJs) of the entomopathogenic nematode (EPN) Heterorhabditis baujardi in laboratory conditions, as well as to investigate aspects related to the biochemistry and histopathology of snails exposed or not to the EPNs during three weeks. The EPN exposure induced significant reductions in the concentrations of glucose, total proteins and glycogen (gonad-digestive gland complex) in the snails during the onset of the infection, with the levels being restored as the infection progresses. These alterations were accompanied by increased hemolymph activities of aminotransferases and lactate dehydrogenase, as well as the concentrations of uric acid after the first and second weeks of the experiment. The histopathological analyses of the exposed snails revealed cell necrosis at the end of the first week, tissue inflammatory reactions one and two weeks after exposure, and degeneration three weeks afterward in comparison with the unexposed snails. Finally, scanning electronic microscopy revealed proliferation of fibrous connective tissue three weeks after exposure. The results indicate that P. columella is susceptible to H. baujardi. The exposure favored the establishment of a negative energy balance, increased the activity of enzymes related to tissue damages and promoted accumulation of nitrogen compounds in the host snails. Additionally, was observed in P. columella exposed to the EPNs, significant tissue lesions, and demonstrated the strong pathogenic potential of H. baujardi, indicating its possible application for biological control of this snail.

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.

Ultraestructural study of effects of alkylphospholipid analogs against nematodes.

Alkylphospholipid analogs were initially developed as anticancer agents and were later found to antiparasitic activity. Miltefosine is the prototype alkylphosphocholine and is the first oral treatment against visceral leishmaniasis. Here we investigated the effects of miltefosine and two ring-substituted alkylphosphocholine derivatives, TCAN26 and TC70, on the viability, morphology, and ultrastructure of the life stages of Caenorhabditis elegans and infective larvae of the parasite Strongyloides venezuelensis. Miltefosine displayed activity against C. elegans adults at low concentrations and was more effective than TCAN26 and TC70. Miltefosine inhibited the hatching of eggs, leading to embryonic lethality, and showed larvicidal activity against C. elegans and S. venezuelensis larvae after 24 h. Mitelfosine also induced alterations in the reproductive system of hermaphrodites, causing vulvar prolapse and general effects in the body wall. Electron microscopy analysis showed that miltefosine induced selective embryonic lethality, leading to cell death. Our results suggest that alkylphospholipid analogs are a potential new alternative for anti-nematode chemotherapy.

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.

Bulimulus tenuissimus (mollusca) as a new potential host of Angiostrogylus cantonensis (nematoda), a histological and metabolic study.

The terrestrial gastropod Bulimulus tenuissimus is widespread in South America. It is an intermediate host of many parasites, but there are no records of infection of this snail by Angiostrongylus cantonensis, despite the occurrence of this parasite and angiostrongyliasis cases in the same areas in which B. tenuissimus occurs. For this reason, it is important investigate the susceptibility of B. tenuissimus to A. cantonensis-infection, since it can be used as intermediate host of A. cantonensis, increasing the list of terrestrial gastropods that infect wild and domestic animals and humans with this parasite. The purpose of this study was to evaluate the susceptibility of B. tenuissimus to experimental infection with L1 larvae of A. cantonensis. The snails were exposed to 1200 L1 larvae and it was possible observe many developing larvae in the cephalopedal mass and mantle tissues, with intense hemocyte infiltration and collagen deposition, but no typical granuloma structures were formed. The glucose content and lactate dehydrogenase activity in the hemolymph varied, indicating an increase of anaerobic energy metabolism in the middle of infection, but with a tendency to return to normal values at the end of pre-patent period. This was corroborated by the marked reduction in the glycogen content in the cephalopedal mass and digestive gland in the first and second week after exposure, followed by a slight increase in the third week. The content of pyruvic acid in the hemolymph was 14.84% lower at the end of pre-patent period, and oxalic acid content was 41.14% higher. These results indicate an aerobic to anaerobic transition process. The PAS reaction showed a large amount of glycogen inside the developing larvae and muscular tissues of the cephalopedal mass, indicating that despite the high consumption of this polysaccharide by the parasite, the snail is able to maintain its energy metabolism based on carbohydrates. The results reveal that B. tenuissimus is a robust host, which can live with the developing larvae of A. cantonensis and overcome the metabolic damages resulting from parasitism.

Unveiling the oxidative metabolism of Achatina fulica (Mollusca: Gastropoda) experimentally infected to Angiostrongylus cantonensis (Nematoda: Metastrongylidae).

For the first time, alterations in the oxidative metabolism of Achatina fulica experimentally infected with different parasite loads of Angiostrongylus cantonensis were determined. For this, the hemolymph activities of lactate dehydrogenase (LDH) and hexokinase and the glucose concentrations in the hemolymph, as well as the polysaccharide reserves in the digestive gland and cephalopedal mass, were assessed. Additionally, the contents of some carboxylic acids in the hemolymph of infected and uninfected snails were determined by high-performance liquid chromatography (HPLC), permitting a better understanding of the alterations related to the host's oxidative metabolism. As the main results, activation of oxidative pathways, such as the glycolytic pathway, was demonstrated in response to the increase in the activity of hexokinase. This tendency was confirmed by the decrease in the contents of glucose in the hemolymph of parasitized snails, indicating that the infection by A. cantonensis alters the host's metabolism, and that these changes are strongly influenced by the parasite load. This metabolic scenario was accompanied by activation of the anaerobic fermentative metabolism, indicated not only by an increase in the activity of (LDH), but also by a reduction of the content of pyruvic acid and accumulation of lactic acid in the hemolymph of parasitized snails. In this circumstance, maintenance of the host's redox balance occurs through activation of the fermentative pathways, and LDH plays a central role in this process. Together, the results indicate that A. cantonensis infection induces activation of the anaerobic metabolism of A. fulica, characterized not only by the accumulation of lactic acid, but also by a reduction in the pyruvic acid and oxalic acid contents in the hemolymph of the infected snails.

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.

Evidence of the Presence of Thyroid Hormones in Achatina fulica Snails.

The objective of this study was to identify thyroid hormones and to examine their putative site of synthesis in Achatina fulica snails. For this purpose, radioimmunoassays were performed for T3 and T4 before and after long starvation with or without hemolymph deproteinization. Sodium/iodide symporter activity in vivo was analyzed through 125I administration with and without KClO4 pretreatment. Only T4 was detected, and its concentration decreased due to starvation or deproteinization. However, high-performance liquid chromatography analysis also showed the presence of T2 and T3 apart from T4, but rT3 was not detected in the A. fulica hemolymph. The sodium/iodide symporter activity was greater in cerebral ganglia than digestive gland, but KClO4 treatment did not inhibit iodide uptake in any of the tissues analyzed. Altogether, our data confirm for the first time the presence of thyroid hormones in A. fulica snails and suggest their participation in the metabolism control in this species, although the putative site of hormone biosynthesis remains to be elucidated.

Long-term clinical follow-up of patients undergoing percutaneous alcohol septal reduction for symptomatic obstructive hypertrophic cardiomyopathy.

BACKGROUND: Alcohol septal ablation (ASA) is an alternative treatment for symptomatic hypertrophic obstructive cardiomyopathy (HOCM) patients refractory to pharmacological therapy. We sought to evaluate the immediate and long-term incidence of death and changes in life quality in a consecutive cohort submitted to ASA. METHODS AND RESULTS: Between October 1998 and December 2013, a total of 56 patients (mean age 53.2 ± 15.5) with symptomatic refractory HOCM were treated with ASA and followed during 15 years (mean 8 ± 4 years). There were 7 (12.5%) deaths, 2 (3.6%) being of cardiac cause. The Kaplan-Meier survival probability estimate was 96.4% at 1 year, 87.7 at 5 years and 81.0% at 12 years post-ASA. Significant improvement was observed in life quality assessed by DASI index and NYHA functional class as well as in the left ventricle outflow tract (LVOT) gradient reduction (from 92.8 ± 3.3 mm Hg to 9.37 ± 6.7 mm Hg, P < 0.001) and septum thickness (from 23.9 ± 0.6 mm to 12.9 ± 1.0 mm, P < 0.001). Only one patient (1.7%) required permanent pacemaker immediately after ASA. During follow-up, one patient had a repeated ASA, three patients underwent myectomy and other four required ICD/pacemaker. In the multivariate model only post-ASA LVOT residual gradient and left ventricle mass were associated with worse prognosis. CONCLUSIONS: In this long-term clinical follow-up without losses, ASA was effective in improving quality of life and NYHA functional class, with relatively low mortality and very low need for immediate permanent pacemaker implantation. © 2016 Wiley Periodicals, Inc.

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

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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.

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 profile of Achatina fulica (Mollusca: Gastropoda) after infection by different parasitic loads of Angiostrongylus cantonensis (Nematoda, Metastrongylidae).

The effect of experimental infection by different parasitic loads of Angiostrongylus cantonensis (Nematode, Metastrongylidae) on the activities of the aminotransferases and concentration of total proteins, uric acid and urea in the hemolymph of Achatina fulica (Mollusca, Gastropoda) were investigated. There was a significant decrease in the concentration of total proteins in the exposed snails to 5000 or more larvae. This change was accompanied by an increase in the concentrations of urea and uric acid in the hemolymph, suggesting a higher rate of deamination of the amino acids. Besides this, variations in the activities of the aminotransferases were also observed, with the highest values recorded in the groups exposed to greater parasite load. These results suggest an increase in the use of total proteins, since there was increased formation of nitrogenous catabolites, in conformity with an increase in the aminotransferase activities. Infection was verified by the fact that L3 larvae recovered from the snails was proportion to the exposure dose of L1 larvae. Histopathological results also indicated presence of an inflammatory cell infiltrate, favoring an increase of both transaminases.

Physiological changes in Rhipicephalus microplus (Acari: Ixodidae) experimentally infected with entomopathogenic fungi.

Carbohydrate metabolism plays an important role in the physiology and maintenance of energy stores within living organisms. However, when organisms are exposed to adverse physiological conditions, such as during pathogenic infection, these organisms begin to use alternative substrates (proteins and lipids) for energy production. This paper studied the carbohydrate metabolism of Rhipicephalus microplus after infection with Beauveria bassiana and Metarhizium anisopliae. The parameters evaluated were glucose concentration, enzymatic activities of lactate dehydrogenase (LDH), alanine aminostransferase (ALT) and aspartate aminostransferase (AST), amounts of uric acid and urea in the hemolymph, and amount of glycogen in the fat body. The results showed changes in nitrogenous products, including an increase in the amount of urea detected 48 h after infection with both fungi. The enzymatic activities of LDH, ALT, and AST were increased after infection. The amount of glucose was increased 24 h after infection with B. bassiana and was reduced 48 h after infection with both fungi. The amount of glycogen in the fat body was reduced at different times of infection with both fungi. These results demonstrate, for the first time, the changes in carbohydrate metabolism of R. microplus after infection with M. anisopliae and B. bassiana and contribute to a better understanding of this host-parasite relationship. Together with knowledge of diseases that affect these ticks and their susceptibility to entomopathogens, an understanding of tick physiology will be necessary for the effective implementation of current biological control methods and will assist in the discovery of new methods to control this ectoparasite.

Changes in the lipid profile of Bradybaena similaris (Férussac, 1821) (Gastropoda, Xanthonychidae) during the development of Eurytrema coelomaticum (Giard and Billet, 1892) (Digenea, Dicrocoeliidae).

The effect of experimental exposure of Bradybaena similaris by Eurytrema coelomaticum on the cholesterol and triglycerides total levels circulating in the hemolymph and the neutral lipids in the digestive gland-gonad (DGG) complex of the host snail were studied. In this study, snails were dissected after one, two, and three months of infection to collect the hemolymph and DGG to measure the cholesterol and triacylglycerol levels in the hemolymph and neutral lipid fractions in the tissues. The results for the hemolymph showed that the infection by this trematode resulted in a significant decrease in the concentrations of triacylglycerol during all periods analyzed, as well as, increase in the levels of cholesterol observed after the second and third months of study. This decrease suggests the possible use of triacylglycerol by both parasite and host as alternative substrate in maintaining their energy metabolisms. In parallel, the increase in the contents of cholesterol results from a severe cellular disorganization in snail, especially DGG, as a consequence of the intense plastic processes showed by parasite. Additionally, changes in the neutral lipid profile in the DGG of the infected snails were observed, indicating the importance of these molecules for successful infection.

Metabolic and histopathological profile of Rattus norvegicus (Wistar) experimentally infected by Angiostrongylus cantonensis (Chen, 1935).

Eosinophilic meningitis is a disease characterized by increased eosinophils in the cerebrospinal fluid (CSF), which is the most commonly caused by invasion of the central nervous system by helminths, as occurs in Angiostrongylus cantonensis infections. The rodent Rattus norvegicus is the definitive natural host and humans act as accidental hosts and can become infected by eating raw or undercooked snails or food contaminated with infective L3 larvae. Recently in Brazil there have been four cases of eosinophilic meningitis due to ingestion of infected Achatina fulica. To evaluate biochemical and histopathological changes caused by this parasite, R. norvegicus were experimentally infected with 100 L3 larvae of A. cantonensis. After the anesthetic procedure, serum from the rodents was collected from the inferior vena cava for evaluation of the levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALKP), gamma-glutamyl transferase (GGT), total protein and its fractions. During the necropsy, the liver was collected and weighed. Then a 1-g fragment was extracted from the major lobe to quantify the hepatic glycogen and fragment remainder was taken from the same lobe and fixed in Milloning's formalin for histopathological examination. Additionally, helminths were collected from the brain and lungs of the rodents. The activities of AST, ALT, ALKP and GGT in the serum and hepatic glycogen increased in response to infection, while the levels of globulin and total protein increased only in the eighth week of infection and there was a reduction in the levels of serum glucose. Albumin and bilirubin concentrations remained stable during the experiment. Infection with A. cantonensis caused metabolic and histopathological changes in the rodents. This study can contribute to a better understanding of the relationship between A. cantonensis and R. norvegicus.

Physiological alterations in Bradybaena similaris (Stylommatophora: Bradybaenidae) induced by the entomopathogenic nematode Heterorhabditis indica (Rhabditida: Heterorhabditidae) strain LPP1.

Heterorhabditis is a nematode found in the soil that is used as an important biological control agent against various organisms. However, few studies have been performed of its use against snails and the present study is the first to investigate the effect of experimental exposure of Bradybaena similaris to Heterorhabditis indica LPP1. Two groups of 16 snails were formed: the control group (not exposed) and the treatment, which was exposed for three weeks to infective juveniles (J3) of H. indica LPP1. The entire experiment was conducted in duplicate, using a total of 64 snails. After this period, the snails were dissected to collect the hemolymph to evaluate the possible physiological alterations, namely total proteins, uric acid and hemolymph urea, as well as the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) as a result of the infection. The terrariums were analyzed on alternate days throughout the experiment to count the dead snails. Intense proteolysis was observed in the infected snails. An increase in the level of uric acid and reduction of the hemolymph urea content indicated that the infection by H. indica results in the inversion of the excretion pattern of the host snail. Variations in the aminotransferase activities were also observed, with the infected group presenting significantly higher values (p<0.05) than the control group for both ALT and AST. The exposure to H. indica LPP1 caused 55% mortality, with the highest rate observed in the first week after exposure (30%). These results suggest that the use of H. indica LPP1 is a feasible alternative for the biological control of B. similaris.

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.

Carbohydrate metabolism evaluation of terrestrial snail Subulina octona (Gastropoda, Subulinidae) experimentally infected by the Paratanaisia bragai digenetic trematode (Digenea, Eucotylidae).

Among the effects of the larval development of digenetic trematodes on their intermediate hosts, changes in the carbohydrate metabolism in the snails stand out. The aim of this study was to analyze, every 10 days after infection (d.p.i.), the effects of Paratanaisia bragai infection on the glycogen content in the digestive gland and cephalopedal mass in Subulina octona snail, and also verify the glucose concentration and the enzyme D- and L-lactate dehydrogenase activity (EC1.1.1.27 and EC1.1.1.28) (LDH) and the concentration of some metabolites(oxalic, succinic, pyruvic and lactic acid) presents in the hemolymph. Histochemical analisys were also performed. We verified a total increase of 54.81% in glucose concentration in infected snails and an oscillating pattern in the glycogen content in the cephalopedal mass and in the digestive gland. LDH activity shows an increase of 10 d.p.i. (+ 74.32%) and 40 d.p.i. (+ 47.81%) and decrease at 20 d.p.i. and 30 d.p.i. The concentrations of oxalic, succinic and pyruvic acids showed significant and progressive reductions; however, lactic acid had a significant increase. Histological and histochemical analysis showed a tissue disorganization in the cephalopedal mass of infected snails and morphological changes in the digestive gland. These results confirm that infection causes metabolic pathway changes in the snails due to activation of an alternative anaerobic pathway for producing energy, indicated by the increased lactic acid content and LDH activity.