Immunolocalization and pathological alterations following Strongyloides venezuelensis infection in the lungs and the intestine of MHC class I or II deficient mice

The present study, investigated the mechanisms involved in the immune responses of Major Histocompatibility Complex class I or class II knockout mice, following Strongyloides venezuelensis infection. Wild-type C57BL/6 (WT), MHC II(-/-) and MHC I(-/-) mice were individually inoculated with 3000 larvae (U) of S. venezuelensis and sacrificed on days 1, 3, 5, 8, 13 and 21 post-infection (p.i.). Samples of blood, lungs and small intestines were collected. The tissue samples were stained with hematoxylineosin for the pathological analysis. The presence of the parasite was demonstrated by immunoperoxidase analysis. MHC II(-/-) mice presented a significantly higher number of adult worms recovered from the small intestine on day 5 p.i. and presented elevated numbers of eggs in the feces. The infection by S. venezuelensis was completely eliminated 13 days after infection in WT as well as in MHC I(-/-) mice. In MHC II(-/-) mice, eggs and adult worms were still found on day 21 p.i., however, there was a significant reduction in their numbers. In the lung, the parasite was observed in MHC I(-/-) on day 1 p.i. and in MHC II(-/-) mice on days 1 and 5 p.i. In the small intestine of WT mice, a larger number of parasites were observed on day 8 p.i. and their absence was observed after day 13 p.i. Through immunohistochemistry analysis, the parasite was detected in the duodenum of WT on days 5 and 8 p.i., and in knockout mice on days 5, 8 and 13 p.i.; as well as in posterior portions of the small intestine in MHC I(-/-) and MHC II(-/-) on day 13 p.i., a finding which was not observed in WT mice. We concluded that immunohistochemistry analysis contributed to a more adequate understanding of the parasite localization in immunodeficient hosts and that the findings aid in the interpretation of immunopathogenesis in Strongyloides infection. (C) 2008 Elsevier B.V. All rights reserved.

Evolutionary transition to freshwater by ancestral marine palaemonids: evidence from osmoregulation in a tide pool shrimp

The transition from marine/brackish waters to freshwater habitats constitutes a severe osmotic and ionic challenge, and successful invasion has demanded the selection of morphological, physiological, biochemical and behavioral adaptations. We evaluated short-term (1 to 12 h exposure) and long-term (5 d acclimation), anisosmotic extracellular (osmolality, [Na(+), Cl(-)]) and long-term isosmotic intracellular osmoregulatory capability in Palaemon northropi, a neotropical intertidal shrimp. F northropi survives well and osmo- and ionoregulates strongly during short- and long-term exposure to 5-45 parts per thousand salinity, consistent with its rocky tide pool habitat subject to cyclic salinity fluctuations, Muscle total free amino acid (FAA) concentrations decreased by 63% in shrimp acclimated to 5%. salinity, revealing a role in hypoosmotic cell volume regulation; this decrease is mainly a consequence of diminished glycine, arginine and proline. Total FAA contributed 31% to muscle intracellular osmolality at 20 parts per thousand, an isosmotic salinity, and decreased to 13% after acclimation to 5 parts per thousand. Gill and nerve tissue FAA concentrations remained unaltered. These tissue-specific responses reflect efficient anisosmotic and anisoionic extracellular regulatory mechanisms, and reveal the dependence of muscle tissue on intracellular osmotic effectors. FAA concentration is higher in P. northropi than in diadromous and hololimnetic palaemonids, confirming muscle FAA concentration as a good parameter to evaluate the degree of adaptation to dilute media. The osmoregulatory capability of P. northropi may reflect the potential physiological capacity of ancestral marine palaemonids to penetrate into dilute media, and reveals the importance of evaluating osmoregulatory processes in endeavors to comprehend the invasion of dilute media by ancestral marine crustaceans.

Measurement of IgE antibodies to shrimp tropomyosin is superior to skin prick testing with commercial extract and measurement of IgE to shrimp for predicting clinically relevant allergic reactions after shrimp ingestion

Background: Shrimp is a frequent cause of food allergy. Tropomyosin is the major allergen in shrimp, and it shares homology to tropomyosins from other crustaceans, dust mites, cockroach, and parasites. Objective: The aim of this study was to determine the value of detection of IgE to shrimp tropomyosin in the diagnosis of shrimp allergy. Methods: We have studied 35 patients with asthma, rhinitis, or both who were sensitized to Dermatophagoides pteronyssinus. All subjects underwent skin prick testing in addition to double-blind, placebo-controlled food challenges (DBPCFC); oral open challenges; or both with shrimp. Measurements of IgE to shrimp and shrimp tropomyosin were carried out by means of CAP and chimeric ELISA, respectively. Results: Oral challenges confirmed the diagnosis of shrimp allergy in 7 patients. IgE measurement to shrimp tropomyosin was positive in 71.4% of the patients with shrimp allergy. Of the 28 patients without shrimp allergy, only 7.1% (2/28) had IgE to shrimp tropomyosin compared with 25% (7/28) who had IgE to shrimp and 35.7% (10/28) who had positive skin prick test responses to shrimp. Sensitivity was similar for all 3 methods (71.4%); in contrast, specificity of IgE to shrimp tropomyosin (92.8%) was greater than that of IgE to shrimp (75%) and skin prick testing (64.2%). With regard to diagnostic efficiency, measurement of IgE to shrimp tropomyosin was superior to measurement of IgE to shrimp and skin prick testing (88.5%, 74.2%, and 65.7%, respectively). Conclusion: Use of measurements of IgE to shrimp tropomyosin provided added value to the diagnosis of shrimp allergy. (J Allergy Clin Immunol 2010;125:872-8.)

Bathymetric distribution of the shrimp Rimapenaeus constrictus (Stimpson, 1874) (Decapoda, Penaeidae) in two locations off the southeastern Brazilian coast

The distribution of benthic organisms is directly or indirectly associated with the physical and chemical properties of the water and sediment. This study analysed the spatial and temporal distribution of Rimapenaeus constrictus in unconsolidated sublittoral sediments of two areas off the northern coast of the state of Sao Paulo, Brazil. We also analysed the association of environmental factors with the occurrence of this species. Shrimp were collected monthly from July 2001 to June 2003, with a fishing boat equipped with two double-rig nets, in the regions of Ubatuba (UBA) and Caraguatatuba (CA). The collections were made during the day, along five transects with mean depths of 5, 10, 15, 20, 25, 30, and 35 m. We obtained a total of 5478 individuals, 3403 (UBA = 2025 and CA = 1378) during the first year, and 2075 (UBA = 875 and CA = 1200) during the second year. Significant differences in abundance were observed in relation to depth, season of the year, and also in the interaction between region and depth. Higher abundances occurred in fall and winter, independently of the sampling year. The largest numbers of shrimp were caught on the 20-m transect in UBA and the 25-m transect in CA. It is concluded that the distribution pattern of this species is closely related to environmental factors, and the temperature of the bottom water and the sediment texture were the most significant variables affecting the distribution.

Sexual System, Sex Ratio, and Group Living in the Shrimp Thor amboinensis (De Man): Relevance to Resource-Monopolization and Sex-Allocation Theories

The sexual system of the symbiotic shrimp Thor amboinensis is described, along with observations on sex ratio and host-use pattern of different populations. We used a comprehensive approach to elucidate the previously unknown sexual system of this shrimp. Dissections, scanning electron microscopy, size-frequency distribution analysis, and laboratory observations demonstrated that T amboinensis is a protandric hermaphrodite: shrimp first mature as males and change into females later in life. Thor amboinensis inhabited the large and structurally heterogeneous sea anemone Stichoclactyla helianthus in large groups (up to 11 individuals) more frequently than expected by chance alone. Groups exhibited no particularly complex social structure and showed male-biased sex ratios more frequently than expected by chance alone. The adult sex ratio was male-biased in the four separate populations studied, one of them being thousands of kilometers apart from the others. This study supports predictions central to theories of resource monopolization and sex allocation. Dissections demonstrated that unusually large males were parasitized by an undescribed species of isopod (family Entoniscidae). Infestation rates were similarly low in both sexes (approximate to 11%-12%). The available information suggests that T. amboinensis uses pure search promiscuity as a mating system. This hypothesis needs to be formally tested with mating behavior observations and field measurements on the movement pattern of both sexes of the species. Further detailed studies on the lifestyle and sexual system of all the species within this genus and the development of a molecular phylogeny are necessary to elucidate the evolutionary history of gender expression in the genus Thor.

Structural changes in the epithelium of the small intestine and immune cell infiltration of enteric ganglia following acute mucosal damage and local inflammation

An acute enteritis is commonly followed by intestinal neuromuscular dysfunction, including prolonged hyperexcitability of enteric neurons. Such motility disorders are associated with maintained increases in immune cells adjacent to enteric ganglia and in the mucosa. However, whether the commonly used animal model, trinitrobenzene sulphonate (TNBS)-induced enteritis, causes histological and immune cell changes similar to human enteric neuropathies is not clear. We have made a detailed study of the mucosal damage and repair and immune cell invasion following intralumenal administration of TNBS. Intestines from untreated, sham-operated and TNBS-treated animals were examined at 3 h to 56 days. At 3 h, the mucosal surface was completely ablated, by 6 h an epithelial covering was substantially restored and by 1 day there was full re-epithelialisation. The lumenal epithelium developed from a squamous cell covering to a fully differentiated columnar epithelium with mature villi at about 7 days. Prominent phagocytic activity of enterocytes occurred at 1-7 days. A surge of eosinophils and T lymphocytes associated with the enteric nerve ganglia occurred at 3 h to 3 days. However, elevated immune cell numbers occurred in the lamina propria of the mucosa until 56 days, when eosinophils were still three times normal. We conclude that the disruption of the mucosal surface that causes TNBS-induced ileitis is brief, a little more than 6 h, and causes a transient immune cell surge adjacent to enteric ganglia. This is much briefer than the enteric neuropathy that ensues. Ongoing mucosal inflammatory reaction may contribute to the persistence of enteric neuropathy.

Immunohistochemical analysis of neuron types in the mouse small intestine

The definition of the nerve cell types of the myenteric plexus of the mouse small intestine has become important, as more researchers turn to the use of mice with genetic mutations to analyze roles of specific genes and their products in enteric nervous system function and to investigate animal models of disease. We have used a suite of antibodies to define neurons by their shapes, sizes, and neurochemistry in the myenteric plexus. Anti-Hu antibodies were used to reveal all nerve cells, and the major subpopulations were defined in relation to the Hu-positive neurons. Morphological Type II neurons, revealed by anti-neurofilament and anti-calcitonin gene-related peptide antibodies, represented 26% of neurons. The axons of the Type II neurons projected through the circular muscle and submucosa to the mucosa. The cell bodies were immunoreactive for choline acetyltransferase (ChAT), and their terminals were immunoreactive for vesicular acetylcholine transporter (VAChT). Nitric oxide synthase (NOS) occurred in 29% of nerve cells. Most were also immunoreactive for vasoactive intestinal peptide, but they were not tachykinin (TK)-immunoreactive, and only 10% were ChAT-immunoreactive. Numerous NOS terminals occurred in the circular muscle. We deduced that 90% of NOS neurons were inhibitory motor neurons to the muscle (26% of all neurons) and 10% (3% of all neurons) were interneurons. Calretinin immunoreactivity was found in a high proportion of neurons (52%). Many of these had TK immunoreactivity. Small calretinin neurons were identified as excitatory neurons to the longitudinal muscle (about 20% of neurons, with ChAT/calretinin/+/- TK chemical coding). Excitatory neurons to the circular muscle (about 10% of neurons) had the same coding. Calretinin immunoreactivity also occurred in a proportion of Type II neurons. Thus, over 90% of neurons in the myenteric plexus of the mouse small intestine can be currently identified by their neurochemistry and shape.

Transepithelial transport of glucose and mRNA of glucose transporters in the small intestine of rats with iron-deficiency anemia

Objective: To evaluate the transepithelial transport of sodium, glucose, potassium, and water and the mRNA level of the sodium-glucose cotransporter (SGLT1) and the facilitated sugar transporter (GLUT2) in the small intestine of iron-deficient rats. Methods: After 6 wk of receiving diets with low or normal iron content, rats (Wistar-EPM) were subjected to two experiments: 1) evaluation of the transepithelial transport of sodium, glucose, potassium, and water by an ""in vivo"" experimental model of intestinal perfusion and 2) determination of relative SGLT1 and GLUT2 mRNA levels in the proximal, intermediate, and distal portions of the small intestine by the northern blotting technique. Results: Hemoglobin and hepatic iron levels were statistically lower in the anemic rats. The mean transepithelial transports of sodium (-33.0 mu Eq . min(-1) . cm(-1)), glucose (426.0 mu M . min(-1) . cm(-1)), and water (0.4 mu L . min(-1) . cm(-1)) in the small intestine of the anemic rats were significantly lower than in the control group (349.1 mu Eq . min(-1) cm(-1), 842.6 mu M . min(-1) . cm(-1), and 4.3 mu l . min(-1) cm(-1), respectively, P < 0.05). The transepithelial transport of potassium was similar for both groups. The relative SGLT1 mRNA levels of the anemic rats in the intermediate (1.796 +/- 0.659 AU) and distal (1.901 +/- 0.766 AU) segments were significantly higher than the values for the control rats (intermediate 1.262 +/- 0.450 AU, distal 1.244 +/- 0.407 AU). No significant difference was observed for the relative SLGT1 mRNA levels in the proximal segment or for the GLUT2 mRNA levels in all segments. Conclusion: Iron deficiency decreases the absorption of glucose, sodium, and water and increases SGLT1 mRNA in the intermediate and distal segments of the small intestine of rats. (C) 2011 Elsevier Inc. All rights reserved.

Protection against high-dose homologous infection in calves immunized with intestine or membrane extracts from Haemonchus placei

Control of Haemonchus placei, one of the most important cattle nematodes in Brazil, relies on the use of anthelmintics. However, there is a need for integrated control, which includes active immunization. The aim of this work was to assess the protection afforded to calves by immunization with adult H. placei extracts against a high-dose challenge infection, a condition frequently found in the tropics. Holstein calves aged 8-10 months were immunized four times with intestinal extracts (Group D) or with a Triton X-100-soluble fraction of adult H. placei (Group A), challenge-infected with 120,000 infective larvae and sacrificed 40 days later. Immunized animals had higher IgG titers than the controls against tested fractions after the 2nd immunization, peaking after the 4th. Sera from both immunized groups recognized bands of similar apparent mass in both antigenic preparations, some of which were similar in molecular weight to Haemonchus contortus antigens with known protective effect to sheep. Egg counts were 49% and 57% lower in Groups A and D than in controls, respectively. High levels of protection were observed in two of the four calves in Group D, as evidenced by very low worm numbers recovered at necropsy, absence of eggs in the uteri of the recovered females and reduced worm length. Group D animals also showed milder signs of anemia than the other infected animals. Results demonstrate that protection against homologous high-dose challenge can be achieved by immunizing calves with H. placei gut antigens. (C) 2007 Elsevier B.V. All rights reserved.

30 million years of Permian volcanism recorded in the Choiyoi igneous province (W Argentina) and their source for younger ash fall deposits in the Parana Basin: SHRIMP U-Pb zircon geochronology evidence

We present four SHRIMP U-Pb zircon ages for the Choiyoi igneous province from the San Rafael Block, central-western Argentina. Dated samples come from the Yacimiento Los Reyunos Formation (281.4 +/- 2.5 Ma) of the Cochico Group (Lower Choiyoi section: andesitic breccias, dacitic to rhyolitic ignimbrites and continental conglomerates). Agua de los Burros Formation (264.8 +/- 2.3 Ma and 264.5 +/- 3.0 Ma) and Cerro Carrizalito Formation (251.9 +/- 2.7 Ma Upper Choiyoi section: rhyolitic ignimbrites and pyroclastic flows) spanning the entire Permian succession of the Choiyoi igneous province. A single ziron from the El Imperial Formation, that is overlain unconformably by the Choiyoi succession, yielded an early Permian age (297.2 +/- 5.3 Ma). while the main detrital zircon population indicated an Ordovician age (453.7 +/- 8.1 Ma). The new data establishes a more precise Permian age (Artinskian-Lopingian) for the section studied spanning 30 Ma of volcanic activity. Volcanological observations for the Choiyoi succession support the occurrence of explosive eruptions of plinian to ultraplinian magnitudes, capable of injecting enormous volumes of tephra in the troposphere-stratosphere. The new SHRIMP ages indicate contemporaneity between the Choyoi succession and the upper part of the Parana Basin late Paleozoic section, from the Irad up to the Rio do Rasto formations, encompassing about 24 Ma. Geochemical data show a general congruence in compositional and tectonic settings between the volcanics and Parana Basin Permian ash fall derived layers of bentonites. Thickness and granulometry of ash fall layers broadly fit into the depletion curve versus distance from the remote source vent of ultraplinian eruptions. Thus, we consider that the Choiyoi igneous province was the source of ash fall deposits in the upper Permian section of the Parana Basin. Data presented here allow a more consistent correlation between tectono-volcanic Permian events along the paleo-Pacific margin of southwestern Gondwana and the geological evolution of neighboring Paleozoic foreland basins in South America and Africa. (C) 2010 International Association for Gondwana Research. Published by Elsevier B.V. All rights reserved.

Contribution of SHRIMP U-Pb zircon geochronology to unravelling the evolution of Brazilian Neoproterozoic fold belts

The South-American continent is constituted of three major geologic-geotectonic entities the homonym platform (consolidated at the end of the Cambrian) the Andean chain (essentially Meso-Cenozoic) and the Patagonian terrains affected by tectonism and magmatism through almost all of the Phanerozoic The platform is constituted by a series of cratonic nuclei (pre-Tonian fragments of the Rodinia fission) surrounded by a complex fabric of Neoproterozoic structural provinces Two major groups of orogenic processes (plate interaction cycles) constitute the evolution of these provinces the older occurred in the Tonian (smaller in area) and the younger Brasiliano that is present in all provinces The Tonian cycles (pre-Rodinia fission?) are still being sorted out and many questions still need to be answered The Brasiliano orogenic collage events (post-Rodinia fission?) developed in three main stages in part coeval from a province to another and are 650-600 580-560 and 540-500 Ma respectively (the late event reaching the Ordovician) The first group of orogenies is recorded in practically all provinces The third group is restricted to part of the Mantiqueira Province (southeast of the platform Buzios Orogeny) and present in the Pampean province (SW of the platform) For all these groups of orogenic events there are considerable records of rock assemblages related to processes of convergent plate interaction opening accretion collision and further extrusion There is a good correlation between the geologic and geotectonic data and geochemical and isotopic data The late tectonic processes (post-orogenic magmatism foreland basins etc) of the first two groups compete in time in distinct spaces with the peak of orogenic processes in the third group The introduction of the SHRIMP U-Pb methodology was fundamental to separate the Tonian and post-Tonian orogenic groups and their respective divisions in time and space Thus there are still many open points/problems which lead to expectations of addressing these issues in the near future with the more Intense use of this methodology (C) 2010 Elsevier B V All rights reserved

Constraining the age of the iporanga formation with SHRIMP U-Pb zircon: Implications for possible ediacaran glaciation in the Ribeira Belt, SE Brazil

The Ribeira belt in SE Brazil is a Neoproterozoic to Early Palaeozoic orogen, whose architecture and history is not yet fully understood. The depositional age of many of the sedimentary sequences in the Ribeira Belt remains unconstrained, and with debate concerning their depositional environment and tectonic setting. In this paper we present SHRIMP zircon U/Pb age constraints for one such problematic unit in the Ribeira Belt the lporanga Formation - and discuss the significance of this age with regards to the timing of Neoproterozoic glacial events in southeast Brazil. Using a felsic volcanic unit immediately under the lporanga Formation and granite cobbles from breccias in its basal parts a reconnaissance SHRIMP U/Pb zircon maximum depositional age of 580 Ma is assigned for the base of this unit. This age is marginally younger than the 625605 Ma ages for intrusions into the Lajeado and Ribeira subgroups, with which the lporanga Formation is in tectonic contact. This indicates that the Lajeado and Ribeira subgroups are not stratigraphically equivalent to the lporanga Formation, as thought previously by some workers. The maximum depositional age of 580 Ma also places a maximum time constraint on the tectonic juxtaposition of the lporanga Formation with other supracrustal units, and on the greenschist facies metamorphism and isoclinal folding that affected it. The potential glacial origin for the lporanga Formation, if correct, would place it in the late Ediacaran - provisionally equivalent to the Gaskiers glaciation. (c) 2007 International Association for Gondwana Research. Published by Elsevier B.V. All rights reserved.

Effects of ascorbic acid supplementation in ileum myenteric neurons of streptozotocin-induced diabetic rats

The exacerbation of the oxidative stress and of the polyol pathway which impair damage myenteric plexus are metabolic characteristics of diabetes. The ascorbic acid (AA) is an antioxidant and an aldose reductase inhibitor, which may act as neuroprotector. The effects of AA supplementation on the density and cellular body profile area (CP) of myenteric neurons in STZ-induced diabetes in rats were assessed. Four groups with five animals each were formed: normoglycemic (C); diabetic (D); AA-treated diabetic (DS) and AA-treated normoglycemic (CS). Dosagen of 50mg of AA were given, three times a week, for each animal (group DS and CS). Ninety days later and after euthanasia, the ileum was collected and processed for the NADPH-diaphorase technique. There were no differences (P>0.05) in the neuronal density among the groups. The CP area was lower (P<0.05) in the DS and CS groups, with a higher incidence of neurons with a CP area exceeding 200µm² for groups C and D. The AA had no influence on the neuronal density in the ileum but had a neuroprotective effect, preventing the increase in the CP area and allowing a higher number of neurons with a CP area with less than 200µm².

Isolamento de Salmonella enterica em gambás (Didelphis aurita e Didelphis albiventris) do Estado de São Paulo, Brasil

No Brasil, não há relato de estudos de Salmonella em gambás, sendo assim, este trabalho tem por objetivo determinar a frequência de isolamento de Salmonella enterica em gambás (D. aurita e D. albiventris) no Estado de São Paulo. No período de janeiro de 2005 a dezembro de 2006, foram necropsiados 106 D. aurita e 40 D. albiventris e colhidos fragmentos de intestinos delgado, grosso e suabe da cloaca. As amostras foram plaqueadas diretamente em ágar Mac Conkey, paralelamente suspendidas nos caldos Rappaport-Vassiliadis e Tetrationato e posteriormente plaqueados em ágar XLT4. As colônias sugestivas de Salmonella foram confirmadas através de provas bioquímicas e sorotipagem. Encontrou-se Salmonella enterica em 17,0% (18/106) dos D. aurita. Destes, 50% apresentaram positividade no intestino delgado (ID), 88,9% no intestino grosso (IG) e 66,7% na cloaca. Da espécie S. enterica, as subespécies encontradas foram: diarizonae (11,1%) houtenae e enterica (5,5% cada um); enquanto da subespécie S. enterica enterica os sorotipos foram Newport (83,3%), Typhimurium e Cerro (5,5% cada um). Nos D. albiventris, 17,5% (7/40) eram positivos, sendo que se encontraram 42,8% no ID, 85,7% no IG e 71,4% na cloaca. O sorotipo mais prevalente também foi Newport (71,4%), seguido por Typhimurium, Bareilly e Thompson (14,3% cada um). Através dos resultados obtidos neste estudo pode-se comprovar a presença de Salmonella enterica no trato intestinal de gambás no Brasil.

Modelo de suprimento sanguíneo do intestino grosso do tamanduá bandeira (Myrmecophaga tridactyla)

Em seis animais adultos Myrmecophaga tridactyla estudou-se o modelo de suprimento sanguíneo do intestino grosso, que é dependente das artérias mesentérica cranial (AMCr) e caudal (AMC). Os espécimes coletados conforme as normas do IBAMA (Instituto Brasileiro do Meio Ambiente e dos Recursos Naturais Renováveis) foram perfundidos com água (40 C), injetados com látex corado, fixados em formol e conservados em solução alcoólica (50%). O mesocólon está disposto na linha sagital mediana e fixa o intestino grosso à parede dorsal do abdome. Derivaram da AMC: a artéria retal cranial, sete a 14 artérias cólicas e uma ou duas artérias ileocólicas, que apresentaram anastomoses de irrigação com a AMCr. A AMC finaliza-se na borda mesocólica das alças intestinais, emitindo ramos cólicos retos a partir das arcadas justacólicas, que penetram na intimidade da musculatura longitudinal. Ao longo do trajeto da AMC foram observadas ilhas arteriais, e a região ileocólica apresentou maior densidade vascular.