Evaluation of immunological, parasitological and molecular methods for the diagnosis of Schistosoma mansoni infection before and after chemotherapy treatment with praziquantel in experimentally infected Nectomys squamipes

In low endemicity areas of schistosomiasis, the recommended diagnostic method of coprological examination results in an underestimation of infection cases. Alternative diagnostic methods have been developed, such as immunodiagnostic and molecular techniques. In this study we evaluated three methods used in the diagnosis of Schistosoma mansoni infection: parasitological (Kato-Katz), immunological (ELISA) and molecular (real time PCR), and also investigated the sensitivity of each technique in the cure determination after treatment with praziquantel using the water rat Nectomys squamipes, a natural reservoir for S. mansoni, as an experimental model. Two infection laboratory experiments were carried out. The first experiment aimed to observe the evolution of the immunological response in the first moments after infection and in the first months after treatment. The second experiment aimed to compare the efficacy of the three diagnostic techniques after infection and after treatment over a more extended time period. In the first experiment, 44% of the infected animals showed IgG reactivity after two weeks of infection, and 94% were positive based on serology 30 days after infection. The serological IgG titers increased just after infection but decreased gradually after treatment. In the second experiment, 89% of the animals showed positive IgG titers 22 days after infection. Only 68% of the animals showed positive results on the coproscopic diagnostic analysis and 79% did so by qPCR, 50 days after infection. Treated animals showed negative results on coproscopy one month after treatment but remained positive by serology even 12 months after treatment, although showing a decline in immunologic reaction after treatment. By qPCR analysis, all animals showed negative results three months after treatment, except for one animal. The parasitosis can be detected by coproscopy only six weeks after infection, and by serology 14 days after infection. The qPCR was a better diagnostic method for confirming the infection cure of S. mansoni. In early infection, this method was less efficient than serology but was slightly more efficient than the Kato-Katz method. We suggest that the methods should be used in low endemic areas as follows: serology should be used in the initial diagnosis in a population with potential positive cases; subsequently, coproscopy should be used in IgG positive cases to confirm the current infection; and qPCR should be used to evaluate the infection cure after treatment and is also a very valuable tool when there are cases showing positive IgG and negative coproscopy.     

Gastroprotective and ulcer-healing mechanisms of ellagic acid in experimental rats

Ellagic acid (EA), a plant-derived polyphenol, exhibits antioxidant, anti-inflammatory, and gastroprotective effects. Its gastroprotective mechanisms have not been fully elucidated nor have its effects on chronic ulcer previously been described. Toward these ends, the antiulcer activities of EA were evaluated in acute (ethanol and indomethacin) and chronic (acetic acid) ulcer models in Wistar rats. In this study, oral administration of EA significantly prevented the gastric ulceration caused by ethanol, indomethacin, and acetic acid treatments. Its gastroprotective mechanism in ethanol-induced ulcer were partly due to intensification in the endogenous production of nitric oxide, an antioxidant effect by replenishing depletion of endogenous nonprotein sulfhydryls and attenuation of tumor necrosis factor-α increase, whereas in indomethacin ulcer, it is partly due to a reduction in the plasma level of leukotriene B(4). In acetic acid ulcer, promotion of ulcer-healing effects was partly due to attenuation of the elevated levels of the inflammatory cytokines TNF-α, interferon-γ, and interleukins-4 and -6. These findings suggest that ellagic acid exerts its antiulcer activity by strengthening the defensive factors and attenuating the offensive factors.     

Carbon,nitrogen and organic C fractions in topsoil affected by conversion from silvopastoral to different land use systems

The conversion of silvopasture to different land use systems cause effective changes in soil carbon distribution, due to disturbances in soil aggregation promoted by soil management and changes in crop residues inputs and decomposability. We evaluate the C and N stocks, and organic C fractions in soils under continuous arable land (AR) and silvopasture with apple trees and grass (SP); and after 4 years of conversion from silvopasture to arable land (SP-AR) and grassland (SP-GL). Total N (TN) and organic C (TOC), as well as microbial biomass carbon (C_MB), light fraction (C_LF) and heavy fraction (C_HF) were evaluated at two different depths (0–10 and 10–20 cm). After 4 years of conversion, SP-AR and SP-GL presented C and N stocks similar to the observed for SP when the 0–20 cm depth was considered. However, AR presented TOC and TN stocks around 21 and 10% lower than SP, respectively. SP-AR tended to present the lowest C_MB stocks and was positively correlated with salt extractable organic C (r ² = 0.60, P < 0.001). C_LF values declined by 62% from 0–10 to the 10–20 cm at SP and SP-GL, however there was no variation with increasing depth for AR and SP-AR. C_HF represented the highest C fraction in soil, corresponding to 82% of TOC. Except for AR, δ¹³C values of the light fraction increased with increasing depth. In general, heavy fraction tended to be more enriched in δ¹³C than light fraction. In a long-term, conventional tillage can significantly contribute to reduce TOC and TN stocks when compared to the silvopastoral system.     

Short-term effects of agroforestry systems on soil health in Southeastern Brazil

Agroforestry Systems - The Soil Management Assessment Framework (SMAF) has successfully been tested as an objective tool to quantify land use and management effects on soil health, including under...     

MUC1 expression in canine malignant mammary tumours and relationship to clinicopathological features

Mucin-1 (MUC1) is over-expressed in human breast carcinomas and is linked to a poorer prognosis. In this study, MUC1 expression in 32 spontaneous canine malignant mammary tumours was characterised in relation to histological type, mode of growth, grade, lymph node metastases and distant metastases. All tumours exhibited immunostaining for MUC1. In the normal canine mammary gland, MUC1 was expressed mainly in the apical cellular membrane, while in carcinomas MUC1 was detected in the cytoplasm only (56.3%) or in the cytoplasm with membrane accentuation (43.7%). There was a significant association between development of distant metastases and MUC1 over-expression (P=0.03), but no association with histological type, histological grade, mode of growth or lymph node metastasis.     

Chinese Box turtle (Cuora flavomarginata) with lymphoid leukemia characterized by immunohistochemical and cytochemical phenotyping

Lymphoid leukemia of T‐cell origin was diagnosed in a male Chinese Box turtle, Cuora flavomarginata, of approximately 25 years of age. The turtle presented with a history of anorexia, open‐mouth breathing, and lethargy for one week. The CBC findings included a mildly increased PCV, and severe leukocytosis due to high numbers of atypical cells interpreted to be blasts. The blasts were medium‐sized cells with round to pleomorphic nuclei, slightly clumped chromatin, indistinct nucleoli, and scant moderate‐to‐dark blue cytoplasm with occasional red‐to‐purple cytoplasmic granulation. Cytochemical and immunohistochemical staining indicated that the neoplastic cells were positive for CD3 and α‐naphthyl butyrate esterase (ANBE), leading to the diagnosis of T‐cell lymphoid leukemia. Histology of tissues collected at necropsy showed multifocal infiltrations of neoplastic round cells in the liver, spleen, kidneys, testicles, pancreas, thyroid, duodenum, bone marrow, epicardium, and myocardium. Transmission electron microscopy failed to identify viral particles within the neoplastic cells. This article describes the hematologic, histologic, and ultrastructural abnormalities associated with lymphoid leukemia in this turtle, and advanced diagnostic methods used for phenotyping the T‐cell origin.     

Bovine vaccinia,a systemic infection: Evidence of fecal shedding,viremia and detection in lymphoid organs

Bovine vaccinia (BV) is a zoonosis caused by Vaccinia virus (VACV) that affects dairy cattle and milkers, causing economic losses and impacting animal and human health. Based on the clinical presentation, BV appears to be a localized disease, with lesions restricted to the skin of affected individuals. However, there are no studies on the pathogenesis of the disease in cows to determine if there is a systemic spread of the virus and if there are different ways of VACV shedding. The objective of this work was to study if there is a systemic spread of VACV in experimentally infected cows and to study the kinetics of VACV circulation in the blood and shedding in the feces of these animals. To this end, eight crossbred lactating cows were used. Three teats of each cow were inoculated with the GP2V strain of VACV. All animals were monitored daily, and blood and fecal samples were collected for 67 days post-infection (dpi). After this period, four of these previously infected cows were immunosuppressed using dexamethasone. Viral DNA was continuously detected and quantified in the blood and feces of these animals in an intermittent way, even after the resolution of the lesions. At slaughter, tissues were collected, and viral DNA was detected and quantified in the mesenteric and retromammary lymph nodes, ileum, spleen and liver. The detection of VACV DNA in the feces for a longer period (67 dpi) and in the lymphatic organs provides new evidence about VACV elimination and suggests that BV could be a systemic infection with a chronic course and viral shedding through the feces.