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1.
Acta Parasitol ; 65(2): 496-503, 2020 Jun.
Article in English | MEDLINE | ID: mdl-32124202

ABSTRACT

INTRODUCTION: A new coccidian species of the genus Eimeria Schneider, 1875 (Apicomplexa: Eimeriidae), is reported from the bat host Myotis riparius Handley from Ilha Grande, a large island off the coast of the State of Rio de Janeiro, in southeastern Brazil. METHODS: Bats were captured in 13 mist nets (10 × 3 m), which were set within the experimental plots, and through active searches of the daytime roosts of Molossus molossus Pallas found in Vila Dois Rios. Containment was made in bags for the collection of feces and identification of coccidia. A survey was conducted on the coccidia species described so far (Table 2). RESULTS: The oöcysts of Eimeria riparii n. sp. are ellipsoidal to cylindroidal with an extremely thin, bi-layered wall, slightly rough. Two polar granules are present, micropyle and oöcyst residuum are both absent. The sporocysts are ellipsoidal, the sporocyst residuum is formed by sparse, rounded granules of varying sizes; the Stieda body is trapezoidal and a sub-Stieda body is absent. Sporozoites are banana shaped. With the new species described here, a total of 40 Eimeria spp. have been described infecting bat hosts, belonging to 30 species of 18 genera and 5 families. CONCLUSION: The subsequent increase in the known diversity of bats has been derived from the ongoing expansion of research in a number of different areas of taxonomy and ecology although the number of studies of the associated coccidian parasites of the family Eimeriidae has increased more slowly.


Subject(s)
Chiroptera/parasitology , Coccidiosis/veterinary , Eimeria/classification , Animals , Brazil , Coccidiosis/parasitology , Eimeria/isolation & purification , Eimeria/ultrastructure , Feces/parasitology , Forests , Intestine, Small/parasitology , Intestine, Small/pathology , Islands , Microscopy, Interference , Oocysts/classification , Oocysts/isolation & purification , Oocysts/ultrastructure
2.
Parasitol Res ; 119(1): 291-298, 2020 Jan.
Article in English | MEDLINE | ID: mdl-31820167

ABSTRACT

We describe three new coccidian species of the genus Eimeria Schneider 1875 (Apicomplexa: Eimeriidae) and redescribe and report Eimeria zygodontomyis Lainson and Shaw, 1990 in the montane grass mouse, Akodon montensis Thomas, 1913 from the Serra dos Órgãos National Park in southeastern Brazil. Sporulated oocysts of Eimeria zygodontomyis are ellipsoidal to cylindrical with a 0.6 (0.5-0.8) µm thick very delicate bi-layered wall; length × width (n = 49) 18.3 × 12.5 (16-20 × 11-13) µm; length/width ratio of 1.4 (1.2-1.6); 1 polar granule occasionally present; micropyle, residuum both absent. Sporocysts are ellipsoidal; length × width 8.5 × 5.2 (8-11 × 5-6) µm; length/width ratio of 1.5 (1.3-1.7) µm; Stieda body is prominent; sub-Stieda body is absent; sporocyst residuum is compact. Sporulated oocysts of Eimeria montensis n. sp. are spheroidal to subspheroidal with a 1.2 (1.0-1.4) µm thick bi-layered wall; outer layer lightly pitted; length × width (n = 30) 16.3 × 12.5 (15-17 × 13-15) µm; length/width ratio of 1.3 (1.0-1.4); 1 polar granule present; micropyle, residuum both absent. Sporocysts are ellipsoidal; length × width 7.2 × 5.1 (6-8 × 4-6) µm; length/width ratio of 1.4 (1.2-1.6); Stieda body is present, sub-Stieda body is absent; sporocyst residuum consists of small, scattered granules. Sporulated oocysts of Eimeria uricanensis n. sp. are ovoidal to pyriform with a 1.4 ( 1.3-1.6) µm thick bi-layered wall; outer layer lightly pitted; length × width (n = 40) 26.6 × 18.6 (23-30 × 17-20) µm; length/width ratio of 1.4 (1.3-1.6); 1 polar granule present; micropyle, residuum both absent. Sporocysts are ellipsoidal, length × width 13.3 × 8.0 (10-16 × 7-9) µm; length/width ratio of 1.7 (1.5-1.9); Stieda body, sub-Stieda body both absent; sporocyst residuum consists of a cluster of granules, forming a spheroid mass. Sporulated oocysts of Eimeria parnasiensis n. sp. are subspheroidal to ellipsoidal with a 1.8 ( 1.3-2.4) µm thick bi-layered wall; outer layer lightly pitted; length × width (n = 54) 28.2 × 21.9 (26-32 × 19-28) µm; length/width ratio of 1.3 (1.2-1.4); 1 polar granule present; micropyle is absent; oocyst residuum is present and consists of a cluster of granules of varying thickness. Sporocysts are ovoidal, tapering towards the Stieda body; length × width 12.2 × 7.6 (10-13 × 6-9) µm; length/width ratio of 1.6 (1.4-1.9); Stieda body is present; sub-Stieda body is absent; sporocyst residuum is present and consists of an aggregate of thin granules.


Subject(s)
Coccidiosis/veterinary , Eimeria/classification , Rodent Diseases/parasitology , Sigmodontinae/parasitology , Animals , Brazil , Coccidiosis/parasitology , Eimeria/cytology , Feces/parasitology , Oocysts/cytology , Parks, Recreational
3.
J Invertebr Pathol ; 170: 107314, 2020 02.
Article in English | MEDLINE | ID: mdl-31866115

ABSTRACT

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.


Subject(s)
Angiostrongylus cantonensis/physiology , Biomphalaria/metabolism , Carbohydrate Metabolism , Echinostoma/physiology , Host-Parasite Interactions , Animals , Coinfection
4.
Exp Parasitol ; 195: 66-70, 2018 Dec.
Article in English | MEDLINE | ID: mdl-30401655

ABSTRACT

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.


Subject(s)
Angiostrongylus cantonensis/physiology , Biomphalaria/parasitology , Echinostoma/physiology , Animals , Biomphalaria/anatomy & histology , Biomphalaria/physiology , Clutch Size , Cricetinae , Disease Reservoirs/parasitology , Disease Vectors , Host-Parasite Interactions , Oviposition , Reproduction , Sigmodontinae/parasitology
5.
Rev Bras Parasitol Vet ; 27(1): 42-51, 2018.
Article in English | MEDLINE | ID: mdl-29641790

ABSTRACT

Helminth parasites have been studied as potential accumulators for different pollutants. Echinostoma paraensei is a foodborne trematode whose vertebrate host, the rodent Nectomys squamipes, is naturally exposed to environmental pesticides. However, little information exists regarding the pesticide's effects on helminths. This study investigated the morphological effects on the trematode, E. paraensei, after experimental Roundup® herbicide exposure, in concentrations below those recommended for agricultural use. After two hours of exposure, scanning electron microscopy (SEM) showed changes to the tegument, such as furrowing, shrinkage, peeling, spines loss on the peristomic collar, and histopathological evidence of altered cells in the cecum and acinus vitelline glands with vacuoles and structural changes to the muscular layers. Glycidic content was decreased, primarily in the connective tissue. As E. paraensei is an intestinal parasite of the semi-aquatic wild rodent, N. squamipes, it is predisposed to pesticide exposure resulting from agricultural practices. Therefore, we emphasize the need to evaluate its impact on helminth parasites, due to their pivotal role in regulating host populations.


Subject(s)
Echinostoma/anatomy & histology , Echinostoma/drug effects , Glycine/analogs & derivatives , Herbicides/pharmacology , Animals , Echinostoma/ultrastructure , Glycine/pharmacology , Microscopy, Electron, Scanning , Glyphosate
6.
Rev. bras. parasitol. vet ; 27(1): 41-50, Jan.-Mar. 2018. tab, graf
Article in English | LILACS | ID: biblio-899316

ABSTRACT

Abstract Helminth parasites have been studied as potential accumulators for different pollutants. Echinostoma paraensei is a foodborne trematode whose vertebrate host, the rodent Nectomys squamipes, is naturally exposed to environmental pesticides. However, little information exists regarding the pesticide's effects on helminths. This study investigated the morphological effects on the trematode, E. paraensei, after experimental Roundup® herbicide exposure, in concentrations below those recommended for agricultural use. After two hours of exposure, scanning electron microscopy (SEM) showed changes to the tegument, such as furrowing, shrinkage, peeling, spines loss on the peristomic collar, and histopathological evidence of altered cells in the cecum and acinus vitelline glands with vacuoles and structural changes to the muscular layers. Glycidic content was decreased, primarily in the connective tissue. As E. paraensei is an intestinal parasite of the semi-aquatic wild rodent, N. squamipes, it is predisposed to pesticide exposure resulting from agricultural practices. Therefore, we emphasize the need to evaluate its impact on helminth parasites, due to their pivotal role in regulating host populations.


Resumo Helmintos parasitos tem sido estudados como acumuladores potenciais para diferentes poluentes. O trematódeo E. paraensei tem como hospedeiro vertebrado o roedor Nectomys squamipes naturalmente exposto a pesticidas no meio ambiente. No entanto, pouca informação está disponível sobre os efeitos dos pesticidas em helmintos parasitos. O presente estudo investigou, em condições experimentais, os efeitos morfológicos no trematódeo E. paraensei após a exposição ao herbicida Roundup®, em concentrações abaixo das recomendadas para a utilização agrícola. A microscopia eletrônica de varredura (MEV) mostrou após duas horas de exposição, alterações no tegumento, como enrugamento, contração e descamação com perda de espinhos no colar peristômico e análise histopatológica evidenciou células do ceco alteradas, as glândulas vitelínicas com vacúolos e mudanças estruturais nas camadas musculares. Diminuição do conteúdo glicídico, principalmente no tecido conjuntivo, também foi observado. Considerando a predisposição à exposição a pesticidas agrícolas de N. squamipes infectado por E. paraensei, são necessários estudos para avaliar o impacto de tais resíduos frente aos helmintos e seus hospedeiros.


Subject(s)
Animals , Echinostoma/anatomy & histology , Echinostoma/drug effects , Glycine/analogs & derivatives , Herbicides/pharmacology , Microscopy, Electron, Scanning , Echinostoma/ultrastructure , Glycine/pharmacology
7.
Parasitol Res ; 117(2): 371-376, 2018 Feb.
Article in English | MEDLINE | ID: mdl-29230581

ABSTRACT

A total of 53 specimens of the montane grass mouse, Akodon montensis Thomas, 1913 were collected in the Serra dos Órgãos National Park (SONP) in November 2014 and July 2015. The fecal material was analyzed, and a prevalence of 7.5% was recorded for a new coccidian species of the genus Eimeria Schneider, 1875, with part of its endogenous development recorded in the small intestine. The oocysts of a new coccidian species of genus Eimeria are ellipsoidal to subspherical. The wall is bi-layered, c. 1.5 µm (1.3-1.6 µm) thick, outer layer rough. Oocyst (n = 126) mean length is 25.3 µm (21.0-28.0 µm), with a width of 20.2 µm (17.0-22.0 µm) and mean length/width (L:W) ratio of 1.3 (1.2-1.4). Polar granule is present, with the oocyst residuum as a large spherical to subspherical globule. Sporocyst shape (n = 126) is ellipsoidal, with a mean length of 11.8 µm (9.3-14.4 µm), width of 7.9 µm (6.7-9.3 µm), and mean L:W ratio of 1.5 (1.4-1.7). Sporocysts with nipple-like Stieda body and sub-Stieda body are absent. A sporocyst residuum formed by several globules, usually along the sporocyst wall. This is the first record of Eimeria in the montane grass mouse from Brazil.


Subject(s)
Coccidiosis/veterinary , Eimeria/classification , Sigmodontinae/parasitology , Animals , Brazil , Coccidiosis/parasitology , Eimeria/isolation & purification , Feces/parasitology , Male , Oocysts
9.
Mem. Inst. Oswaldo Cruz ; 112(5): 328-338, May 2017. tab, graf
Article in English | LILACS | ID: biblio-841797

ABSTRACT

BACKGROUND Angiostrongylus costaricensis is a nematode that causes human abdominal angiostrongyliasis, a disease found mainly in Latin American countries and particularly in Brazil and Costa Rica. Its life cycle involves exploitation of both invertebrate and vertebrate hosts. Its natural reservoir is a vertebrate host, the cotton rat Sigmodon hispidus. The adult worms live in the ileo-colic branches of the upper mesenteric artery of S. hispidus, causing periarteritis. However, there is a lack of data on the development of vasculitis in the course of infection. OBJECTIVE To describe the histopathology of vascular lesions in S. hispidus following infection with A. costaricensis. METHODS Twenty-one S. hispidus were euthanised at 30, 50, 90 and 114 days post-infection (dpi), and guts and mesentery (including the cecal artery) were collected. Tissues were fixed in Carson’s Millonig formalin, histologically processed for paraffin embedding, sectioned with a rotary microtome, and stained with hematoxylin-eosin, resorcin-fuchsin, Perls, Sirius Red (pH = 10.2), Congo Red, and Azan trichrome for brightfield microscopy analysis. FINDINGS At 30 and 50 dpi, live eggs and larvae were present inside the vasa vasorum of the cecal artery, leading to eosinophil infiltrates throughout the vessel adventitia and promoting centripetal vasculitis with disruption of the elastic layers. Disease severity increased at 90 and 114 dpi, when many worms had died and the intensity of the vascular lesions was greatest, with intimal alterations, thrombus formation, iron accumulation, and atherosclerosis. CONCLUSION In addition to abdominal angiostrongyliasis, our data suggest that this model could be very useful for autoimune vasculitis and atherosclerosis studies.


Subject(s)
Animals , Arteritis/parasitology , Arteritis/pathology , Strongylida Infections/complications , Strongylida Infections/pathology , Atherosclerosis/pathology , Angiostrongylus , Rodentia , Time Factors , Sigmodontinae , Disease Models, Animal
10.
Mem Inst Oswaldo Cruz ; 112(5): 328-338, 2017 May.
Article in English | MEDLINE | ID: mdl-28443978

ABSTRACT

BACKGROUND: Angiostrongylus costaricensis is a nematode that causes human abdominal angiostrongyliasis, a disease found mainly in Latin American countries and particularly in Brazil and Costa Rica. Its life cycle involves exploitation of both invertebrate and vertebrate hosts. Its natural reservoir is a vertebrate host, the cotton rat Sigmodon hispidus. The adult worms live in the ileo-colic branches of the upper mesenteric artery of S. hispidus, causing periarteritis. However, there is a lack of data on the development of vasculitis in the course of infection. OBJECTIVE: To describe the histopathology of vascular lesions in S. hispidus following infection with A. costaricensis. METHODS: Twenty-one S. hispidus were euthanised at 30, 50, 90 and 114 days post-infection (dpi), and guts and mesentery (including the cecal artery) were collected. Tissues were fixed in Carson's Millonig formalin, histologically processed for paraffin embedding, sectioned with a rotary microtome, and stained with hematoxylin-eosin, resorcin-fuchsin, Perls, Sirius Red (pH = 10.2), Congo Red, and Azan trichrome for brightfield microscopy analysis. FINDINGS: At 30 and 50 dpi, live eggs and larvae were present inside the vasa vasorum of the cecal artery, leading to eosinophil infiltrates throughout the vessel adventitia and promoting centripetal vasculitis with disruption of the elastic layers. Disease severity increased at 90 and 114 dpi, when many worms had died and the intensity of the vascular lesions was greatest, with intimal alterations, thrombus formation, iron accumulation, and atherosclerosis. CONCLUSION: In addition to abdominal angiostrongyliasis, our data suggest that this model could be very useful for autoimune vasculitis and atherosclerosis studies.


Subject(s)
Angiostrongylus , Arteritis/parasitology , Atherosclerosis/parasitology , Strongylida Infections/complications , Animals , Arteritis/pathology , Atherosclerosis/pathology , Disease Models, Animal , Rodentia , Sigmodontinae , Strongylida Infections/pathology , Time Factors
11.
Vet Parasitol ; 228: 144-152, 2016 Sep 15.
Article in English | MEDLINE | ID: mdl-27692317

ABSTRACT

Nematode parasites of the cardiopulmonary system of livestock and pet animals have been receiving attention due to the pathogenic effects they produce in the course of the infection. However, parasitism in wild animals by metastrongilid nematodes has been neglected, resulting in potential risk to wildlife. Heterostrongylus heterostrongylus is the etiological agent of bronchial pneumonia in the black-eared opossum, Didelphis aurita, a widely distributed marsupial frequently reported to inhabit areas from wild environments to peri-urban spaces. In this study, we reassessed the taxonomy of H. heterostrongylus, describing and comparing morphology of L1 and L3 larvae and adult worms with closely related angiostrongilids, and inferring phylogenetic affinities within the family Angiostrongylidae; we also detailed histopathological reactions under natural infection. Ultrastructural morphology and light microscopy confirmed cephalic structures with well-developed trilobed lips, patterns of caudal bursa rays, spicules shape, terminal anal aperture and presence of the ventral protuberance in L3 larvae, characteristics present for the family Angiostrongylidae. Our molecular phylogenetic analyses, based on the partial small subunit (SSU) 18S rRNA gene, suggested that H. heterostrongylus and Didelphostrongylus hayesi are closely related. Pathological analyses agreed with previous findings, showing that cardiopulmonary parasitism provokes verminous pneumonia and mucous bronchiolitis with hypertrophy of the smooth bronchiolar muscle. These data confirmed that H. heterostrongylus represents a pulmonary pathogen in D. aurita, produces severe pathological effects, and may represent risk to domestic and wild animals.


Subject(s)
Didelphis/parasitology , Metastrongyloidea/classification , Strongylida Infections/veterinary , Animals , DNA, Helminth/chemistry , DNA, Helminth/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Female , Lung/parasitology , Lung/pathology , Male , Metastrongyloidea/genetics , Metastrongyloidea/isolation & purification , Metastrongyloidea/ultrastructure , Microscopy, Electron, Scanning/veterinary , Phylogeny , Sequence Analysis, DNA , South America , Strongylida Infections/diagnosis , Strongylida Infections/parasitology
12.
Exp Parasitol ; 134(2): 228-34, 2013 Jun.
Article in English | MEDLINE | ID: mdl-23541880

ABSTRACT

Parasitic castration in the snail-trematode relationship can be understood as any change in the reproductive function of the snail that is due to interference by the developing larvae inside the snail that leads to the reduction or complete disruption of egg-laying activity. This study was designed to observe the parasitic castration of Biomphalaria glabrata infected with Schistosoma mansoni during both the pre-patent and patent periods. The effect of infection on snail fecundity and fertility, growth rate and survival was studied during the 62 days following miracidia exposure. An integrated approach was employed that used biochemical and histological tools over the same period. To study the effect of infection on reproduction, we individually exposed 30 snails to 5 miracidia each and tracked their fertility and fecundity. For our histopathological studies, 50 snails were exposed to 20 miracidia each, and for our histochemical studies, 50 snails were exposed to 5 miracidia each. An equal number of uninfected snails were used as a control for each group. The B. glabrata exposed to the BH strain of S. mansoni showed 50% positivity for cercarial shedding. Both the experimental and control groups showed 100% survival. The pre-patent period lasted until 39 days after exposure to miracidia. Exposed snails that showed cercarial shedding exhibited higher growth rates than either exposed snails that did not demonstrate cercarial shedding or uninfected controls. Exposed snails without cercarial shedding and uninfected controls showed no differences in the reproductive parameters evaluated during the patent period; snails experiencing cercarial shedding showed a reduction in fecundity and fertility. These snails began to lay eggs only after the 50th day post miracidia exposure. The haemolymph glucose levels showed an oscillating pattern that decreased during periods of greater mobilisation of energy by the larvae and was accompanied by a depletion of glycogen in the cephalopodal mass and digestive gland. Histopathological examination at 55 days showed that the ovotestis was highly atrophied. There was almost complete disappearance of germ cells, and the supporting stroma formed a nearly empty net. At day 45, the infected digestive gland showed a high cylindrical epithelium with little preserved cytoplasm. The contents of the secretory granules of the albumen gland of infected animals stained with Alcian blue (AB), pH 1.0, indicating the presence of sulphated carbohydrates. Thus, parasitic castration in the B. glabrata-S. mansoni model may be regulated directly and indirectly by the developmental stage of the trematode and the biochemical and histopathological alterations during the patent period of infection.


Subject(s)
Biomphalaria/parasitology , Schistosoma mansoni/physiology , Animals , Biomphalaria/growth & development , Biomphalaria/physiology , Feces/parasitology , Fertility , Galactans/metabolism , Glucose/metabolism , Glycogen/metabolism , Glycoproteins/metabolism , Hemolymph/chemistry , Humans , Mice , Schistosoma mansoni/isolation & purification
13.
Mem Inst Oswaldo Cruz ; 107(6): 752-9, 2012 Sep.
Article in English | MEDLINE | ID: mdl-22990964

ABSTRACT

Angiostrongylus costaricensis is a nematode that causes abdominal angiostrongyliasis, a widespread human parasitism in Latin America. This study aimed to characterize the protease profiles of different developmental stages of this helminth. First-stage larvae (L1) were obtained from the faeces of infected Sigmodon hispidus rodents and third-stage larvae (L3) were collected from mollusks Biomphalaria glabrata previously infected with L1. Adult worms were recovered from rodent mesenteric arteries. Protein extraction was performed after repeated freeze-thaw cycles followed by maceration of the nematodes in 40 mM Tris base. Proteolysis of gelatin was observed by zymography and found only in the larval stages. In L3, the gelatinolytic activity was effectively inhibited by orthophenanthroline, indicating the involvement of metalloproteases. The mechanistic class of the gelatinases from L1 could not be precisely determined using traditional class-specific inhibitors. Adult worm extracts were able to hydrolyze haemoglobin in solution, although no activity was observed by zymography. This haemoglobinolytic activity was ascribed to aspartic proteases following its effective inhibition by pepstatin, which also inhibited the haemoglobinolytic activity of L1 and L3 extracts. The characterization of protease expression throughout the A. costaricensis life cycle may reveal key factors influencing the process of parasitic infection and thus foster our understanding of the disease pathogenesis.


Subject(s)
Angiostrongylus/enzymology , Proteolysis , Angiostrongylus/classification , Animals , Feces/parasitology , Female , Larva/enzymology , Male , Sigmodontinae
14.
Mem. Inst. Oswaldo Cruz ; 107(6): 752-759, set. 2012. ilus, tab
Article in English | LILACS | ID: lil-649490

ABSTRACT

Angiostrongylus costaricensis is a nematode that causes abdominal angiostrongyliasis, a widespread human parasitism in Latin America. This study aimed to characterize the protease profiles of different developmental stages of this helminth. First-stage larvae (L1) were obtained from the faeces of infected Sigmodon hispidus rodents and third-stage larvae (L3) were collected from mollusks Biomphalaria glabrata previously infected with L1. Adult worms were recovered from rodent mesenteric arteries. Protein extraction was performed after repeated freeze-thaw cycles followed by maceration of the nematodes in 40 mM Tris base. Proteolysis of gelatin was observed by zymography and found only in the larval stages. In L3, the gelatinolytic activity was effectively inhibited by orthophenanthroline, indicating the involvement of metalloproteases. The mechanistic class of the gelatinases from L1 could not be precisely determined using traditional class-specific inhibitors. Adult worm extracts were able to hydrolyze haemoglobin in solution, although no activity was observed by zymography. This haemoglobinolytic activity was ascribed to aspartic proteases following its effective inhibition by pepstatin, which also inhibited the haemoglobinolytic activity of L1 and L3 extracts. The characterization of protease expression throughout the A. costaricensis life cycle may reveal key factors influencing the process of parasitic infection and thus foster our understanding of the disease pathogenesis.


Subject(s)
Animals , Female , Male , Angiostrongylus/enzymology , Proteolysis , Angiostrongylus/classification , Feces/parasitology , Larva/enzymology , Sigmodontinae
15.
Lab Invest ; 89(10): 1140-51, 2009 Oct.
Article in English | MEDLINE | ID: mdl-19721415

ABSTRACT

One difficulty in studying dengue virus (DENV) is the lack of an experimental model that reproduces the human disease. In a previous work, we have shown that BALB/c mice intraperitoneally inoculated with a DENV-2 isolate presented viremia and mild focal areas of liver injuries. In this study, mice were inoculated by the intravenous route and presented extensive damage areas in the liver tissue, which were evaluated by histopathological and ultrastructural analysis. Hepatic injury was noted mainly around the central vein and portal tracts. Damages consist of hepatocyte injury, including steatosis, swelling and necrosis. Further, erythrophagocytosis, intercellular edema and vascular damages were evident, including hemorrhage, which is characteristic of the dengue-induced hepatitis in human liver. Hepatic lesions were already noted 2 days post infection (p.i.), although effects were more extensive after the seventh day p.i. An increase in alanine aminotransferase and aspartate aminotransferase serum levels was detected 7 and 14 days p.i., respectively, and had correlation to hepatic lesions. Alterations caused by the DENV infection were self-limiting, with a remarkable reduction of all liver damages 49 days p.i. Virus antigens were detected in hepatocytes, Kupffer cells and vascular endothelium, suggesting virus replication in these cells. In situ hybridization, using a probe that anneals in the virus negative RNA strand, showed positive reaction in hepatocytes and vascular endothelium cells of infected mice, thus confirming virus replication in such cells. In general, results revealed that this mouse model reproduces some histopathological effects observed in humans and supports previous findings indicating virus replication in the hepatic tissue.


Subject(s)
Dengue Virus/physiology , Dengue/pathology , Liver/ultrastructure , RNA, Viral/analysis , Virus Replication , Alanine Transaminase/blood , Animals , Antigens, Viral/analysis , Aspartate Aminotransferases/blood , Dengue/blood , Dengue/virology , Disease Models, Animal , Humans , Liver/enzymology , Male , Mice , Mice, Inbred BALB C
16.
Mem Inst Oswaldo Cruz ; 100(4): 407-20, 2005 Jul.
Article in English | MEDLINE | ID: mdl-16113890

ABSTRACT

Angiostrongylus costaricensis lives in the cecal and mesenteric arteries of its vertebrate hosts, and causes an inflammatory disease in humans. To investigate unknown aspects of the abdominal angiostrogyliasis pathogenesis, infected Sigmodon hispidus were sequentially studied in different times of infection. The study revealed that L3 goes alternatively through two migratory courses during its development into an adult worm: lymphatic/venous-arterial and venous portal pathways. The former is considered the principal one, because it is used by most of the larvae. Like other metastrongylides, A. costaricensis passes over the pulmonary circulation to migrate from the lymphatic system to the arterial circulation, where they circulate during some days before reaching their definitive habitat. The oviposition by mature females began on 15th day. Eggs and L1 were detected mainly in the intestine and stomach, surrounded by inflammatory reaction constituted by macrophages, monocytes, and eosinophils. They were also spread to the lungs, mesenteric lymph nodes, pancreas, spleen, and kidneys. The larvae (L1) exhibited the centripetal capacity to invade the lymphatic and venous vessels of the intestine and mesentery. Adult worms that developed in the venous intrahepatic pathway migrated downstream to reach the mesenteric veins and laid eggs that embolized in the portal hepatic vessels.


Subject(s)
Angiostrongylus/physiology , Rodent Diseases/parasitology , Strongylida Infections/parasitology , Angiostrongylus/growth & development , Animals , Disease Models, Animal , Female , Life Cycle Stages , Male , Rodent Diseases/pathology , Sigmodontinae , Strongylida Infections/pathology , Time Factors
17.
Mem. Inst. Oswaldo Cruz ; 100(4): 407-420, July 2005. ilus
Article in English | LILACS | ID: lil-405997

ABSTRACT

Angiostrongylus costaricensis lives in the cecal and mesenteric arteries of its vertebrate hosts, and causes an inflammatory disease in humans. To investigate unknown aspects of the abdominal angiostrogyliasis pathogenesis, infected Sigmodon hispidus were sequentially studied in different times of infection. The study revealed that L3 goes alternatively through two migratory courses during its development into an adult worm: lymphatic/venous-arterial and venous portal pathways. The former is considered the principal one, because it is used by most of the larvae. Like other metastrongylides, A. costaricensis passes over the pulmonary circulation to migrate from the lymphatic system to the arterial circulation, where they circulate during some days before reaching their definitive habitat. The oviposition by mature females began on 15th day. Eggs and L1 were detected mainly in the intestine and stomach, surrounded by inflammatory reaction constituted by macrophages, monocytes, and eosinophils. They were also spread to the lungs, mesenteric lymph nodes, pancreas, spleen, and kidneys. The larvae (L1) exhibited the centripetal capacity to invade the lymphatic and venous vessels of the intestine and mesentery. Adult worms that developed in the venous intrahepatic pathway migrated downstream to reach the mesenteric veins and laid eggs that embolized in the portal hepatic vessels.


Subject(s)
Animals , Male , Female , Angiostrongylus/growth & development , Rodent Diseases/parasitology , Strongylida Infections/parasitology , Disease Models, Animal , Life Cycle Stages , Rodent Diseases/pathology , Sigmodontinae , Strongylida Infections/pathology , Time Factors
18.
Mem. Inst. Oswaldo Cruz ; 99(7): 717-720, Nov. 2004. tab
Article in English | LILACS | ID: lil-391600

ABSTRACT

The association between worm infections and bacterial diseases has only recently been emphasized. This study examined the effect of experimental Angiostrongylus costaricensis infection on endogenous intestinal flora of Swiss Webster mice. Eight mice aging six weeks were selected for this experiment. Four were infected with A. costaricensis and the other four were used as controls. Twenty eight days after the worm infection, all mice in both groups were sacrificed and samples of the contents of the ileum and colon were obtained and cultured for aerobic and anaerobic bacteria. In the mice infected with A. costaricensis there was a significant increase in the number of bacteria of the endogenous intestinal flora, accompanied by a decrease in the number of Peptostreptococcus spp. This alteration in the intestinal flora of mice infected by the nematode may help to understand some bacterial infections described in humans.


Subject(s)
Animals , Mice , Angiostrongylus , Colon , Ileum , Strongylida Infections , Colon , Disease Models, Animal , Ileum
19.
Mem Inst Oswaldo Cruz ; 99(7): 717-20, 2004 Nov.
Article in English | MEDLINE | ID: mdl-15654427

ABSTRACT

The association between worm infections and bacterial diseases has only recently been emphasized. This study examined the effect of experimental Angiostrongylus costaricensis infection on endogenous intestinal flora of Swiss Webster mice. Eight mice aging six weeks were selected for this experiment. Four were infected with A. costaricensis and the other four were used as controls. Twenty eight days after the worm infection, all mice in both groups were sacrificed and samples of the contents of the ileum and colon were obtained and cultured for aerobic and anaerobic bacteria. In the mice infected with A. costaricensis there was a significant increase in the number of bacteria of the endogenous intestinal flora, accompanied by a decrease in the number of Peptostreptococcus spp. This alteration in the intestinal flora of mice infected by the nematode may help to understand some bacterial infections described in humans.


Subject(s)
Angiostrongylus , Intestines/microbiology , Intestines/parasitology , Strongylida Infections/parasitology , Angiostrongylus/immunology , Animals , Bacteria, Aerobic/isolation & purification , Bacteria, Anaerobic/isolation & purification , Colon/immunology , Colon/microbiology , Colon/parasitology , Disease Models, Animal , Ileum/immunology , Ileum/microbiology , Ileum/parasitology , Male , Mice , Strongylida Infections/immunology
20.
Rev. Soc. Bras. Med. Trop ; 36(6): 743-745, nov.-dez. 2003. tab
Article in English | LILACS, Sec. Est. Saúde SP | ID: lil-355340

ABSTRACT

Samples of Achatina fulica were experimentally infected with Angiostrongylus costaricensis larvae, etiological agent of abdominal angiostrongyliasis, showing that A. fulica is susceptible to the parasite. Achatina fulica may be a risk to urbanization of abdominal angiostrongyliasis presumably due to its high proliferation, continuous dispersion and remarkable adaptation in several Brazilian towns.


Subject(s)
Animals , Mice , Angiostrongylus cantonensis , Mollusca , Brazil , Disease Vectors , Host-Parasite Interactions
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