RESUMO
Coiling patterns of heligmonellid nematodes were examined for 520, 208, and 33 individuals of Nippostrongylus brasiliensis, Orientostrongylus tenorai, and Sabanema sp., respectively, collected from murine rodents of Indonesia. Besides typical sinistral coiling, complete dextral coiling was found in 3.3% of N. brasiliensis and 12.1% of Sabanema sp. Mixed coiling with partial sinistral and dextral patterns was also observed in 38.8% of N. brasiliensis, 60.7% of Sabanema sp., and 3.4% of O. tenorai. In dextral coils, the left ventral area with large ridges was located inside as in sinistral coils, keeping the ability to cling to intestinal villi. The cuticular dilatation at left to left dorsal area was located caudally in sinistral coils but rostrally in dextral coils. Presence of mixed coiling indicates that the coiling patterns can change. As the transition of coiling pattern accompanies a change in direction of coil axis, it is surmised that the dextral coiling may be chosen when a worm leaves a villus to move to another villus.
Assuntos
Heligmosomatoidea/fisiologia , Intestinos/parasitologia , Microvilosidades/parasitologia , Murinae/parasitologia , Doenças dos Roedores/parasitologia , Animais , Feminino , Heligmosomatoidea/ultraestrutura , Indonésia , Intestinos/ultraestrutura , Masculino , Microvilosidades/ultraestrutura , Nippostrongylus/fisiologia , Nippostrongylus/ultraestrutura , Ratos/parasitologiaRESUMO
Members of the genus Cryptosporidium are frequent protozoan pathogens in humans and a wide range of animals. There is no consistently effective treatment against cryptosporidiosis, especially in immunodeficient patients. The present study was carried out to study the therapeutic effects of curcumin against cryptosporidiosis in immunosuppressed BALB/c mice. Mice were divided into five groups and immunosuppressed by dexamethasone. Three groups were inoculated with C. parvum oocysts, administered with curcumin, paromomycin, and without treatment. The reminders were regarded as controls. The oocysts in the fecal smear were counted daily. At days 0, 3, 7, and 11 post-treatment, the mice were sacrificed, and the efficacy of drugs was evaluated by comparing the histopathological alterations in jejunum and ileum, measuring the total antioxidant capacity, and malondialdehyde in the affected tissues. The infection was completely eliminated in the curcumin-treated group, and oocyst shedding stopped with no recurrence after drug withdrawal. On the contrary, paromomycin was unable to eliminate C. parvum infection completely, and oocyst shedding continued even 10 days after the drug withdrawal. Based on these findings, curcumin can be a trustworthy compound for the elimination of infection in immunosuppressed hosts. Further evaluation to find its accurate mechanism of action should be considered.
Assuntos
Antiprotozoários/uso terapêutico , Criptosporidiose/tratamento farmacológico , Cryptosporidium parvum/efeitos dos fármacos , Curcumina/uso terapêutico , Animais , Antioxidantes/metabolismo , Antiprotozoários/farmacologia , Bovinos , Criptosporidiose/imunologia , Criptosporidiose/patologia , Cryptosporidium parvum/crescimento & desenvolvimento , Cryptosporidium parvum/fisiologia , Curcumina/farmacologia , Modelos Animais de Doenças , Fezes/parasitologia , Feminino , Íleo/parasitologia , Íleo/patologia , Terapia de Imunossupressão , Jejuno/parasitologia , Jejuno/patologia , Camundongos , Camundongos Endogâmicos BALB C , Microvilosidades/parasitologia , Microvilosidades/patologia , Oocistos/fisiologia , Oxidantes/metabolismo , Paromomicina/farmacologia , Paromomicina/uso terapêutico , Distribuição AleatóriaRESUMO
Cryptosporidium is a genus of protozoal parasites that affects the gastrointestinal epithelium of a variety of hosts. Several models of experimental infection have been described to study the susceptibility, infectivity and pathogenicity among different Cryptosporidium species and isolates. This study aimed to establish an experimental infection of Cryptodporidium canis in canids. Infectivity and pathogenicity have been measured by evaluating the clinical status, pattern of oocyst excretion and histological examination. Results showed that C. canis was not infective for immunocompetent dogs or mice with severe combined immunodeficiency syndrome (SCID). Oocysts were first detected in the feces of immunosuppressed dogs on day 3 post-infection (p.i.), with levels peaking twice on days 10 and 17 p.i. during the patent period. cryptosporidial developmental stages were found in the duodenum and jejunum of dogs in histological sections stained with hematoxylin and eosin (H & E) and using scanning electron microscopy (SEM). Histopathological changes in the intestinal tract of infected dogs were characterized by epithelial metaplasia and dilatation; the integrity of intestinal mucosal epithelial cells was distinctly damaged with whole sheets of cilia sloughed away. Ultrastructural observation data were consistent with histological observations. Based on these findings, the canine model described in this work will be useful to evaluate clinical, parasitological and histological aspects of C. canis infection and will be useful for the further understanding of cryptosporidiosis, drug development, and vaccine development.
Assuntos
Criptosporidiose/parasitologia , Modelos Animais de Doenças , Cães , Hospedeiro Imunocomprometido , Animais , Criptosporidiose/patologia , Cryptosporidium/isolamento & purificação , Cryptosporidium/ultraestrutura , Diarreia/parasitologia , Duodeno/parasitologia , Duodeno/patologia , Duodeno/ultraestrutura , Fezes/parasitologia , Jejuno/parasitologia , Jejuno/patologia , Jejuno/ultraestrutura , Camundongos , Camundongos SCID , Microscopia Eletrônica de Varredura , Microvilosidades/parasitologia , Microvilosidades/patologia , Microvilosidades/ultraestrutura , Oocistos/isolamento & purificaçãoRESUMO
Endogenous stages of the life cycle of Eimeria melanomytis, infecting the peripheral epithelial cells of villi of the small intestine of experimentally infected young dusky rice rats, Melanomys caliginosus , were studied. Giemsa-stained mucosal scrapings and histological sections were examined for all the stages. Eimeria melanomytis has 3 generations of meronts (M), different in size, shape, and number of merozoites (m); and in size, shape, and location of the nuclei within the cytoplasm of the meronts. The 3 meront types, M1-M3, respectively, had 11-14 (m1), 7-10 (m2), and 20-30 (m3) merozoites. Macrogametocytes and microgametocytes, as well as macrogametes and microgametes, complete the sexual cycle forming the unsporulated oocysts. This parasite's endogenous development produced severe intestinal lesions in experimentally infected dusky rice rats.
Assuntos
Coccidiose/veterinária , Eimeria/crescimento & desenvolvimento , Estágios do Ciclo de Vida , Doenças dos Roedores/parasitologia , Sigmodontinae/parasitologia , Animais , Coccidiose/parasitologia , Costa Rica , Eimeria/isolamento & purificação , Eimeria/fisiologia , Fezes/parasitologia , Intestino Delgado/parasitologia , Intestino Delgado/ultraestrutura , Merozoítos , Microvilosidades/parasitologia , Oocistos , Esporos de Protozoários , TrofozoítosRESUMO
After oral infection, Toxoplasma gondii invades intestinal cells, induces breakdown of intestinal physiology and barrier functions, and causes intestinal pathology in some animal species. Although parasites' invasion into host cells is a known phenomenon, the effects of T. gondii infection in the intestinal barrier are still not well established. To evaluate morphological and physiological modifications on the colorectal adenocarcinoma-derived Caco-2 cell line during T. gondii infection, microvilli, tight junction integrity, and transepithelial electrical resistance (TEER) were investigated under infection. It was observed that the dextran uptake (endocytosis) and distribution were smaller in infected than in noninfected Caco-2 cells. The infection leads to the partial loss of microvilli at the cell surface. Claudin-1, zonula occludens-1 (ZO-1), and occludin expressions were colocalized by immunofluorescence and presented discontinuous net patterns in infected cells. Immunoblotting analysis at 24 hr postinfection revealed decreasing expression of occludin and ZO-1 proteins, whereas claudin-1 presented similar expression level compared with noninfected cells. T. gondii decreased TEER in Caco-2 cells 24 hr after infection. Our results suggest that T. gondii infection may lead to the loss of integrity of intestinal mucosa, resulting in impaired barrier function.
Assuntos
Mucosa Intestinal/parasitologia , Toxoplasma/fisiologia , Citoesqueleto de Actina/ultraestrutura , Células CACO-2 , Polaridade Celular , Claudina-1/metabolismo , Dextranos/metabolismo , Impedância Elétrica , Endocitose , Humanos , Mucosa Intestinal/metabolismo , Mucosa Intestinal/ultraestrutura , Microvilosidades/metabolismo , Microvilosidades/parasitologia , Microvilosidades/ultraestrutura , Ocludina/metabolismo , Junções Íntimas/metabolismo , Junções Íntimas/parasitologia , Junções Íntimas/ultraestrutura , Proteína da Zônula de Oclusão-1/metabolismoRESUMO
Giardia lamblia is a protistan parasite that infects and colonizes the small intestine of mammals. It is widespread and particularly endemic in the developing world. Here we present a detailed structural study by 3-D negative staining and cryo-electron tomography of a unique Giardia organelle, the ventral disc. The disc is composed of a regular array of microtubules and associated sheets, called microribbons that form a large spiral, held together by a myriad of mostly unknown associated proteins. In a previous study we analyzed by cryo-electron tomography the central microtubule portion (here called disc body) of the ventral disc and found a large portion of microtubule associated inner (MIPs) and outer proteins (MAPs) that render these microtubules hyper-stable. With this follow-up study we expanded our 3-D analysis to different parts of the disc such as the ventral and dorsal areas of the overlap zone, as well as the outer disc margin. There are intrinsic location-specific characteristics in the composition of microtubule-associated proteins between these regions, as well as large differences between the overall architecture of microtubules and microribbons. The lateral packing of microtubule-microribbon complexes varies substantially, and closer packing often comes with contracted lateral tethers that seem to hold the disc together. It appears that the marginal microtubule-microribbon complexes function as outer, laterally contractible lids that may help the cell to clamp onto the intestinal microvilli. Furthermore, we analyzed length, quantity, curvature and distribution between different zones of the disc, which we found to differ from previous publications.
Assuntos
Microscopia Crioeletrônica/métodos , Citoesqueleto/ultraestrutura , Tomografia com Microscopia Eletrônica/métodos , Giardia lamblia/ultraestrutura , Microtúbulos/ultraestrutura , Trofozoítos/ultraestrutura , Animais , Giardia lamblia/citologia , Giardia lamblia/fisiologia , Giardíase/parasitologia , Interações Hospedeiro-Parasita , Imageamento Tridimensional/métodos , Intestinos/citologia , Intestinos/parasitologia , Intestinos/ultraestrutura , Microvilosidades/parasitologia , Microvilosidades/ultraestrutura , Trofozoítos/fisiologiaRESUMO
Accompanying acute hepatopancreatic necrosis disease (AHPND) in cultivated Asian shrimp has been an increasing prevalence of vermiform, gregarine-like bodies within the shrimp hepatopancreas (HP) and midgut. In high quantity they result in white fecal strings and a phenomenon called white feces syndrome (WFS). Light microscopy (LM) of squash mounts and stained smears from fresh HP tissue revealed that the vermiform bodies are almost transparent with widths and diameters proportional to the HP tubule lumens in which they occur. Despite vermiform appearance, they show no cellular structure. At high magnification (LM with 40-100x objectives), they appear to consist of a thin, outer membrane enclosing a complex of thicker, inter-folded membranes. Transmission electron microscopy (TEM) revealed that the outer non-laminar membrane of the vermiform bodies bore no resemblance to a plasma membrane or to the outer layer of any known gregarine, other protozoan or metazoan. Sub-cellular organelles such as mitochondria, nuclei, endoplasmic reticulum and ribosomes were absent. The internal membranes had a tubular sub-structure and occasionally enclosed whole B-cells, sloughed from the HP tubule epithelium. These internal membranes were shown to arise from transformed microvilli that peeled away from HP tubule epithelial cells and then aggregated in the tubule lumen. Stripped of microvilli, the originating cells underwent lysis. By contrast, B-cells remained intact or were sloughed independently and whole from the tubule epithelium. When sometimes engulfed by the aggregated, transformed microvilli (ATM) they could be misinterpreted as cyst-like structures by light microscopy, contributing to gregarine-like appearance. The cause of ATM is currently unknown, but formation by loss of microvilli and subsequent cell lysis indicate that their formation is a pathological process. If sufficiently severe, they may retard shrimp growth and may predispose shrimp to opportunistic pathogens. Thus, the cause of ATM and their relationship (if any) to AHPND should be determined.
Assuntos
Apicomplexa/fisiologia , Sistema Digestório/patologia , Fezes/parasitologia , Hepatopâncreas/patologia , Microvilosidades/patologia , Penaeidae/parasitologia , Animais , Sistema Digestório/parasitologia , Sistema Digestório/ultraestrutura , Células Epiteliais/parasitologia , Células Epiteliais/patologia , Células Epiteliais/ultraestrutura , Hepatopâncreas/parasitologia , Hepatopâncreas/ultraestrutura , Microscopia Eletrônica , Microscopia Eletrônica de Transmissão , Microvilosidades/parasitologia , Infecções por Protozoários/parasitologia , SíndromeRESUMO
The current treatments for cryptosporidiosis are ineffective, and there is an urgent need to search for more effective and safer alternatives. One such alternative may be treatments derived from natural resources. The pomegranate peel has been used effectively in traditional medicine to cure diarrhea and dysentery. The purpose of this study was to examine the effectiveness of a Punica granatum (pomegranate) peel suspension as a treatment for Cryptosporidium parvum infection. In this study, the effects of this treatment on the ultrastructure of both the intestinal epithelial layer of infected nursling mice and the parasite were observed with a transmission electron microscope. The histological study focused on the examination of the microvilli, columnar epithelium, goblet cells, lamina propria, and crypts of Lieberkuhn. Examination of the ileums of infected mice that received the pomegranate peel suspension demonstrated that the general structure of the ileal tissue of these mice was similar to that of the control group. In the infected mice treated with the suspension, but not the infected/untreated mice, there was an improvement in all ultrastructure aspects at 28days post-inoculation. The study of the ultrastructure of the parasite (C. parvum) in mice treated with the suspension showed that there was decomposition in the parasite to the extent that in some cases we were unable to identify the stage of the parasite due to the severe degeneration. Significant decomposition of the nutrition organ was also observed. Additionally, microgamonte and macrogamonte were not observed in the suspension-treated group, explaining the disappearance of the sexual phases of the parasite in the lumens of this group. In all, this examination demonstrated the restoration of the normal structures of villi and the disappearance of acute symptoms in the suspension-treated mice and showed that the suspension directly affected the parasite at various stages of its development and led to its decomposition and death.
Assuntos
Criptosporidiose/patologia , Cryptosporidium parvum , Íleo/ultraestrutura , Lythraceae/química , Animais , Bovinos , Cryptosporidium parvum/ultraestrutura , Modelos Animais de Doenças , Feminino , Frutas/química , Íleo/parasitologia , Camundongos , Microscopia Eletrônica de Transmissão , Microvilosidades/parasitologia , Microvilosidades/ultraestrutura , Gravidez , SuspensõesRESUMO
Some water-borne protozoan parasites induce diseases through their membrane-associated functional structures and virulence factors that hijack the host cellular molecules and signalling pathways leading to structural and functional lesions in the intestinal barrier. In this Microreview we analyse the insights on the mechanisms of pathogenesis of Entamoeba intestinalis, Giardia and Cryptosporidium observed in the human colon carcinoma fully differentiated colon cancer cell lines, cell subpopulations and clones expressing the structural and functional characteristics of highly specialized fully differentiated epithelial cells lining the intestinal epithelium and mimicking structurally and functionally an intestinal barrier.
Assuntos
Permeabilidade da Membrana Celular/fisiologia , Colo/fisiopatologia , Colo/parasitologia , Neoplasias do Colo/fisiopatologia , Neoplasias do Colo/parasitologia , Linhagem Celular Tumoral , Colo/patologia , Neoplasias do Colo/patologia , Cryptosporidium/patogenicidade , Cryptosporidium/fisiologia , Entamoeba/patogenicidade , Entamoeba/fisiologia , Giardia/patogenicidade , Giardia/fisiologia , Interações Hospedeiro-Parasita/fisiologia , Humanos , Mucosa Intestinal/parasitologia , Mucosa Intestinal/patologia , Mucosa Intestinal/fisiopatologia , Microvilosidades/parasitologia , Microvilosidades/patologia , Microvilosidades/fisiologiaRESUMO
Giardiasis is one of the most common parasitic diseases worldwide, and the disease is an important cause of diarrhoea and malabsorption in children and immunosuppressed individuals. However, there is no evidence that characterises malnutrition as an aggravating factor for this disease. We evaluated changes in villi structures to examine the association between malnutrition and Giardia lamblia infection. We used 32 gerbils, divided into 4 groups: Control (CT) and Control Infected (CTIn), which each received a 20% protein diet, Malnourished (MN) and Malnourished Infected (MNIn), which each received a 5% protein diet. Groups CTIn and MNIn were inoculated with 1×10(6) trophozoites of G. lamblia, while the remaining groups were mock infected. Seven days post-infection, all groups were sacrificed, and the proximal portions of the small intestines were collected for the analysis of villus height, mucus area and extent of Giardia infection. Gerbils fed with a low-protein diet had significantly lower body weights. Malnourished infected animals presented significantly increased production of mucus, suggesting a synergism occurs between malnutrition and Giardiasis, potentially to control the adhesion of Giardia in the mucosa. Villus height was significantly lower in group MNIn compared to CTIn. This work suggests that malnutrition contributes to severity of Giardiasis by decreasing the intestinal absorption capacity via shortening of the villi.
Assuntos
Giardíase/complicações , Giardíase/patologia , Intestino Delgado/patologia , Desnutrição Proteico-Calórica/complicações , Desnutrição Proteico-Calórica/patologia , Animais , Feminino , Gerbillinae , Células Caliciformes/metabolismo , Células Caliciformes/patologia , Mucosa Intestinal/metabolismo , Mucosa Intestinal/parasitologia , Mucosa Intestinal/patologia , Intestino Delgado/metabolismo , Intestino Delgado/parasitologia , Microvilosidades/metabolismo , Microvilosidades/parasitologia , Microvilosidades/patologia , Muco/metabolismoRESUMO
The Cryptosporidium in the small intestine of domestic mice (Mus musculus) was initially described as Cryptosporidium parvum. Recent genetic and biologic characterization of Cryptosporidium isolates indicate that domestic mice are infected with several morphologically indistinguishable intestinal Cryptosporidium parasites with different host specificities, including C. parvum sensu stricto, mouse genotype I, and mouse genotype II. In this study, the morphological, biological, and genetic characteristics of the Cryptosporidium mouse genotype I are described. As a full re-description of C. parvum was made in 1985 for isolates from calves and humans and the name C. parvum has been widely used for the parasite that is infectious to both ruminants and humans, the mouse genotype I is named as Cryptosporidium tyzzeri. Oocysts of the new species (4.64±0.05 µm ×4.19±0.06 µm, with a mean shape index of 1.11±0.02; n=69) are slightly smaller than those of the re-described C. parvum. The prepatent period was six and seven days, and the patent period was 24-28 and 28-29 days in neonatal and adult mice, respectively. Oocysts were not infectious to lambs and calves. Light, transmission electron and scanning electron microscopy studies of the new species showed the presence of developmental stages in the microvillar brush border of the jejunum and ileum of experimentally infected mice, with the infection most intensive in the ileum. It had nucleotide sequences significantly different from C. parvum at the small subunit rRNA, 70 kDa heat shock protein, oocyst wall protein, actin, and the 60 kDa glycoprotein genes. Based on the morphological, genetic, and biological data and in compliance of established Cryptosporidium species naming criteria, this geographically widespread parasite is named as a new species in honor of Ernest Edward Tyzzer, who pioneered Cryptosporidium research.
Assuntos
Criptosporidiose/veterinária , Cryptosporidium/classificação , Camundongos/parasitologia , Doenças dos Roedores/parasitologia , Animais , Animais Recém-Nascidos , Bovinos , Criptosporidiose/parasitologia , Criptosporidiose/transmissão , Cryptosporidium/genética , Cryptosporidium/ultraestrutura , Fezes/parasitologia , Feminino , Genótipo , Especificidade de Hospedeiro , Intestino Delgado/parasitologia , Masculino , Camundongos Endogâmicos BALB C , Microscopia Eletrônica de Varredura/veterinária , Microscopia Eletrônica de Transmissão/veterinária , Microvilosidades/parasitologia , Oocistos/ultraestrutura , Doenças dos Roedores/transmissão , OvinosRESUMO
The study describes the in vivo activity of Lactobacillus casei in malnourished Giardia lamblia-infected BALB/c mice. By experimentation, it was found that daily administration of the probiotic 7 days before inoculation with Giardia trophozoites in malnourished mice efficiently reduced both the severity and duration of giardiasis. More specifically, excretion of Giardia cysts and trophozoites counts were reduced, while faecal lactobacilli counts increased significantly in probiotic-fed malnourished mice, compared with control mice. Interestingly, it was also observed that oral feeding of the probiotic to malnourished mice abrogated all the anthropometric and biochemical anomalies. Histologically, morphological and cellular alteration of microvillus membrane integrity revealed that probiotic administration ameliorated the mucosal damage in malnourished, probiotic-inoculated, Giardia-infected mice compared with the severe microvillus atrophy, Ådematous and vacuolated epithelial cells, and ileitis in malnourished Giardia-infected mice. The results clearly show the antigiardial effect of the probiotic in vivo by modulating the gut cells to inhibit the colonization and multiplication of Giardia trophozoites, thus reducing the severity and duration of murine giardiasis.
Assuntos
Giardia lamblia/crescimento & desenvolvimento , Giardíase/terapia , Lacticaseibacillus casei/fisiologia , Desnutrição/parasitologia , Probióticos/uso terapêutico , Animais , Peso Corporal , Modelos Animais de Doenças , Fezes/microbiologia , Fezes/parasitologia , Giardia lamblia/patogenicidade , Giardíase/sangue , Giardíase/prevenção & controle , Mucosa Intestinal/microbiologia , Mucosa Intestinal/parasitologia , Mucosa Intestinal/patologia , Jejuno/microbiologia , Jejuno/parasitologia , Jejuno/patologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Microvilosidades/microbiologia , Microvilosidades/parasitologia , Microvilosidades/patologia , Trofozoítos/crescimento & desenvolvimentoRESUMO
We assessed the mucosal response of previously infected hamsters to low-dose challenge with the hookworm, Ancylostoma ceylanicum. Hamsters were assigned to five treatment groups (Groups 1-5, respectively): naïve, controls; uninterrupted primary infection from day 0; infected, but treated with anthelmintic on day 35 p.i.; challenge control group given only the second infection on day 63; infected initially, cleared of worms and then challenged. Animals were culled on days 73 and 94 (10 and 31 days after challenge), but additional animals were culled from Group 5 on days 80 and 87. The results showed that villus height declined markedly and progressively over time after challenge in Group 5, whilst depth of the Crypts of Lieberkühn and number of mitotic figures in the crypts increased. Mucosal mast cell numbers were only marginally higher than those in naïve controls and not as high as those in mice with uninterrupted infections. Goblet cell counts showed a major increase, as did eosinophils in relation to naïve controls. Paneth cells were also elevated, but did not change over the course of the experiment. The results also drew attention to the tremendous resilience of hookworms, some adult worms surviving throughout, despite highly inflamed intestines.
Assuntos
Ancylostoma/imunologia , Ancilostomíase/imunologia , Ancilostomíase/patologia , Mucosa Intestinal/imunologia , Mucosa Intestinal/patologia , Vacinação , Imunidade Adaptativa , Ancilostomíase/parasitologia , Ancilostomíase/terapia , Animais , Anti-Helmínticos/uso terapêutico , Cricetinae , Eosinófilos/imunologia , Eosinófilos/parasitologia , Eosinófilos/patologia , Feminino , Interações Hospedeiro-Parasita/imunologia , Humanos , Inflamação/imunologia , Inflamação/parasitologia , Inflamação/patologia , Mucosa Intestinal/parasitologia , Mastócitos/imunologia , Mastócitos/parasitologia , Mastócitos/patologia , Camundongos , Microvilosidades/imunologia , Microvilosidades/parasitologia , Microvilosidades/patologia , Índice Mitótico , Fatores de TempoRESUMO
To examine the infection kinetics and development of alterations in the small intestine of gerbils (Meriones unguiculatus), 72 gerbils were divided into six groups (A to F), with A serving as control and the others inoculated with increasing doses of trophozoites from Giardia duodenalis human isolate. The infection kinetics and the development of histopathological alterations were monitored by optical scanning electron microscopy (SEM). A 12-day prepatent period was observed, with intermittent elimination up to day 35 after inoculation. Statistically significant differences were found between the mean number of trophozoites recovered, per group, on the days of sacrifice, and a positive correlation between the inoculum dosage and the number of trophozoites recovered. Morphometrically, the villus:crypt ratio showed a drop in all the groups when compared with the control group. SEM revealed an increase in mucus production in the inoculated animals and the presence of trophozoite clusters at the top and base of the villi. The dosage of trophozoite inoculum does not interfere in the ability for infection to occur or in the development of histopathological alterations generated by intestinal colonization.
Assuntos
Giardia lamblia/patogenicidade , Giardíase/patologia , Intestino Delgado/patologia , Animais , Modelos Animais de Doenças , Feminino , Gerbillinae , Giardíase/parasitologia , Mucosa Intestinal/parasitologia , Mucosa Intestinal/patologia , Mucosa Intestinal/ultraestrutura , Intestino Delgado/parasitologia , Intestino Delgado/ultraestrutura , Cinética , Masculino , Microscopia Eletrônica de Varredura , Microvilosidades/parasitologia , Microvilosidades/patologia , Microvilosidades/ultraestrutura , VirulênciaRESUMO
Cryptosporidium parvum is a parasitic protozoa increasingly appreciated as a cause of intestinal malabsorptive syndrome leading to malnutrition and/or growth failure. Because a major mechanism for apical peptide absorption by small intestine is via the proton-coupled transporter PepT1, we investigated the expression and functionality of this transporter in our model of acute cryptosporidiosis. Four-day-old Sprague-Dawley rats were inoculated by gavage with 5 x 10(5) oocysts of C. parvum and killed at Day 12 (peak of the infection) or Day 21 (spontaneous clearance of the parasite). PepT1 expression and functionality were quantified in the distal small intestine, preferential site of C. parvum implantation, and in the proximal small intestine, free of parasite, using Western blot and Ussing chambers, respectively. No difference in total PepT1 protein expression or in glycyl-sarcosine fluxes was observed in C. parvum-infected rats compared with controls either on Day 12 or on Day 21, both in the proximal and in the distal small intestine. However, a significant decrease of apical membrane protein expression of PepT1 was observed in C. parvum-infected enterocytes compared with controls. This maintained dipeptide transport observed despite villous atrophy and decreased expression of the protein at the brush-border membrane strongly suggest a transient upregulation of PepT1 activity, probably related to gamma-interferon regulation.
Assuntos
Criptosporidiose/metabolismo , Cryptosporidium parvum/crescimento & desenvolvimento , Simportadores/metabolismo , Animais , Animais Lactentes , Western Blotting , Criptosporidiose/sangue , Criptosporidiose/parasitologia , Dipeptídeos/metabolismo , Enterócitos/metabolismo , Enterócitos/parasitologia , Íleo/metabolismo , Íleo/parasitologia , Interferon gama/sangue , Interferon gama/metabolismo , Mucosa Intestinal/metabolismo , Mucosa Intestinal/parasitologia , Microvilosidades/metabolismo , Microvilosidades/parasitologia , Transportador 1 de Peptídeos , Ratos , Ratos Sprague-Dawley , Regulação para CimaRESUMO
Studies were carried out to identify proteins involved in the interface of Trypanosoma cruzi with the perimicrovillar membranes (PMM) of Rhodnius prolixus. Video microscopy experiments demonstrated high level of adhesion of T. cruzi Dm 28c epimastigotes to the surface of posterior midgut cells of non-treated R. prolixus. The parasites however were unable to attach to gut cells obtained from decapitated or azadirachtin-treated insects. The influence of carbohydrates on the adhesion to insect midgut was confirmed by inhibition of parasite attachment after midgut incubation with N-acetylgalactosamine, N-acetylmannosamine, N-acetylglucosamine, D-galactose, D-mannose or sialic acid. We observed that hydrophobic proteins in the surface of epimastigotes bind to polypeptides with 47.7, 45.5, 44, 43, 40.5, 36, 31 and 13kDa from R. prolixus PMM and that pre-incubation of lectins specifically inhibited binding to 31, 40.5, 44 and 45.5kDa proteins. We suggest that glycoproteins from PMM and hydrophobic proteins from epimastigotes are important for the adhesion of the parasite to the posterior midgut cells of the vector.
Assuntos
Insetos Vetores/parasitologia , Glicoproteínas de Membrana/metabolismo , Rhodnius/parasitologia , Trypanosoma cruzi/fisiologia , Animais , Western Blotting , Carboidratos/farmacologia , Adesão Celular/fisiologia , Eletroforese em Gel de Poliacrilamida , Humanos , Interações Hidrofóbicas e Hidrofílicas , Inseticidas/farmacologia , Mucosa Intestinal/parasitologia , Limoninas/farmacologia , Masculino , Microscopia de Vídeo , Microvilosidades/química , Microvilosidades/parasitologiaRESUMO
T lymphocyte-mediated pathogenesis is common to a variety of enteropathies, including giardiasis, cryptosporidiosis, bacterial enteritis, celiac's disease, food anaphylaxis, and Crohn's disease. In giardiasis as well as in these other disorders, a diffuse loss of microvillous brush border, combined or not with villus atrophy, is responsible for disaccharidase insufficiencies and malabsorption of electrolytes, nutrients, and water, which ultimately cause diarrheal symptoms. Other mucosal changes may include crypt hyperplasia and increased infiltration of intra-epithelial lymphocytes. Recent studies using models of giardiasis have shed new light on the immune regulation of these abnormalities. Indeed, experiments using an athymic mouse model of infection have found that these epithelial injuries were T cell-dependent. Findings from further research indicate that that the loss of brush border surface area, reduced disaccharidase activities, and increase crypt-villus ratios are mediated by CD8+ T cells, whereas both CD8+ and CD4+ small mesenteric lymph node T cells regulate the influx of intra-epithelial lymphocytes. Future investigations need to characterize the CD8+ T cell signaling cascades that ultimately lead to epithelial injury and malfunction in giardiasis and other malabsorptive disorders of the intestine.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Giardíase/patologia , Mucosa Intestinal/patologia , Animais , Giardíase/imunologia , Humanos , Mucosa Intestinal/imunologia , Mucosa Intestinal/parasitologia , Microvilosidades/parasitologia , Microvilosidades/patologiaRESUMO
Alkaline phosphatase (IAP) is a marker of intestinal microvillus membrane. Changes in IAP activity have been studied as a function of Giardia lamblia (G. lamblia) infection using rat as the experimental model. At day 11 and 15 post-infection, enzyme activity was reduced (p<0.01) compared to controls. The enzyme levels were essentially similar to control values by day 30 post-infection. Analysis of the enzyme activity in cell fractions across crypt-villus axis revealed a marked decrease in enzyme activity in the villus tip and mid villus regions but a considerable increase (p<0.01) in enzyme activity in the crypt base of 11 day post-infected animals compared to that in controls. The observed changes in IAP activity were confirmed by assaying the enzyme activity in acrylamide gels using bromo-chloro-indolyl phosphate staining under non-denaturing conditions. These findings indicate differential changes across the crypt-villus axis, but impaired alkaline phosphatase levels in G. lamblia infected rat intestine.
Assuntos
Fosfatase Alcalina/biossíntese , Regulação Enzimológica da Expressão Gênica , Giardia lamblia/fisiologia , Giardíase/enzimologia , Intestinos/enzimologia , Fosfatase Alcalina/genética , Animais , Eletroforese em Gel de Poliacrilamida , Intestinos/parasitologia , Masculino , Microvilosidades/enzimologia , Microvilosidades/parasitologia , Ratos , Ratos WistarRESUMO
T lymphocyte-mediated pathogenesis is common to a variety of enteropathies, including giardiasis, cryptosporidiosis, bacterial enteritis, celiac's disease, food anaphylaxis, and Crohn's disease. In giardiasis as well as in these other disorders, a diffuse loss of microvillous brush border, combined or not with villus atrophy, is responsible for disaccharidase insufficiencies and malabsorption of electrolytes, nutrients, and water, which ultimately cause diarrheal symptoms. Other mucosal changes may include crypt hyperplasia and increased infiltration of intra-epithelial lymphocytes. Recent studies using models of giardiasis have shed new light on the immune regulation of these abnormalities. Indeed, experiments using an athymic mouse model of infection have found that these epithelial injuries were T cell-dependent. Findings from further research indicate that that the loss of brush border surface area, reduced disaccharidase activities, and increase crypt-villus ratios are mediated by CD8+ T cells, whereas both CD8+ and CD4+ small mesenteric lymph node T cells regulate the influx of intra-epithelial lymphocytes. Future investigations need to characterize the CD8+ T cell signaling cascades that ultimately lead to epithelial injury and malfunction in giardiasis and other malabsorptive disorders of the intestine.
Assuntos
Animais , Humanos , /imunologia , /imunologia , Giardíase/patologia , Mucosa Intestinal/patologia , Giardíase/imunologia , Mucosa Intestinal/imunologia , Mucosa Intestinal/parasitologia , Microvilosidades/parasitologia , Microvilosidades/patologiaRESUMO
Cryptosporidium parvum mainly invades the intestinal epithelium and causes watery diarrhea in humans and calves. However, the invasion process has not yet been clarified. In the present study, the invasion process of C. parvum in severe combined immunodeficiency (SCID) mice was examined. Infected mice were necropsied; the ilea were double-fixed routinely and observed by scanning and transmission electron microscopy. In addition, the microvillus membrane was observed by ruthenium red staining. Scanning electron micrographs showed elongation of the microvilli at the periphery of the parasite. The microvilli were shown to be along the surface of the parasite in higher magnification. Transmission electron microscopy confirmed that the invading parasites were located among microvilli. Parasites existed in the parasitophorous vacuole formed by the microvillus membrane. The parasite pellicle attached to the host cell membrane at the bottom of the parasite, and then the pellicle and host cell membrane became unclear. Subsequently, the pellicle became complicated and formed a feeder organelle. In addition, invasion of the parasite was not observed in either a microvillus or the cytoplasm of the host cell. Therefore, C. parvum invades among microvilli, is covered with membranes derived from numerous microvilli, and develops within the host cell.