Molecular and serological detection of Babesia spp. in neotropical and exotic carnivores in Brazilian zoos

Large and small piroplasms have been observed in the blood smears of various wild carnivores, but few studies utilizing molecular characterization have been done. The goal of this present study was to investigate the presence of Babesia sp. by molecular and serologic techniques in exotic and neotropical carnivores maintained in captivity at Brazilian zoos. Blood and sera samples were collected from 146 Brazilian wild felids, 21 exotic felids, 1 genet (Genetta tigrina), 3 European wolves (Canis lupus), and 94 Brazilian wild canids in Brazilian zoos in the S?o Paulo and Mato Grosso states and in the Federal District. A total of 53 wild felids (31.74%) and 10 wild canids (10.31%) were seropositive for Babesia canis by Indirect Fluorescent Antibody Test (IFAT). Antibodies were detected in ocelots, little-spotted cats, margays, pampas cats, jaguars, pumas, jaguarundis, crab-eating foxes, and bush dogs. Babesia sp. DNA, with high similarity to B. leo, was detected in one pampas cat and one genet.     

Phosphate Fertilizers for Sugarcane Used at Pre-Planting (Phosphorus Fertilizer Application)

Phosphate applications are still rare in sugarcane cultivation and can be done with phosphorus fertilizers of residual over. This study aims to evaluate the agronomic efficiency of phosphate fertilization before sugarcane planting and its effect on sugarcane yield. The study was carried out over three crop cycles at the Jalles Machado sugar-mill in Goianésia, GO, Brazil. The treatments consisted of five different phosphorus sources (soluble and insoluble) applied at a rate of 300 kg ha^?1 phosphorus pentoxide (P₂O₅). The phosphorus fertilizers used were: triple superphosphate, mono-ammonium phosphate (MAP), Arad rock phosphate, Itafós rock phosphate, and magnesium term phosphate. The effects of these fertilizers were evaluated by plant height (ground to Top Visible Dewlap; TVD), leaf phosphorus content, agronomic efficiency, and sugarcane yield from cane-plants, first ratoon, and second ratoon. Magnesium term phosphate and triple superphosphate resulted in greater plant-cane height. Leaf phosphorus concentrations were highest with applications of MAP, magnesium term phosphate, and triple superphosphate. Phosphate applications did not affect sugarcane plant yield, but did increase ratoon yield. In general, the residual effect of insoluble phosphate increased over the years. The agronomic efficiency (AE) of phosphate was (in descending order): triple superphosphate (100%), term phosphate (89%), MAP (80%), Itafós rock phosphate (67%), and Arad rock phosphate (60%).     

A simplified spectrophotometric method for routine analysis of saccharin in commercial noncaloric sweeteners

A simple, rapid, and sensitive spectrophotometric method for routine analysis of saccharin in commercial noncaloric sweeteners is proposed. This method is based on the reaction of saccharin with tetrachloro-p-benzoquinone (p-chloranil) accelerated by hydrogen peroxide and conducted in an ethanol:acetone (4:1) medium, producing a violet-red compound (lambda(max) = 550 nm). Beer's law is obeyed in a concentration range of 2.05 x 10(-4) to 3.00 x 10(-3) M with an excellent correlation coefficient (r = 0.9998). The detection limit was 1.55 x 10(-5) M, and the effect of interferences on the spectrophotometric measurements was evaluated. The proposed procedure was applied successfully to the determination of saccharin in noncaloric sweeteners. Recoveries were within 99.2-104.3% with standard deviations ranging from to 0.5-1.6%. Results of the proposed method compare very favorably with those given by the high-performance liquid chromatography method recommended by the Food and Drug Administration.     

Methoxylated xanthones in the quality control of small centaury (Centaurium erythraea) flowering tops.

In the course of a phytochemical study of the bitter tonic plant, small centaury (Centaurium erythraea), six methoxylated xanthones (1,5-hydroxy-3-methoxyxanthone, 1-hydroxy-3,5,6-trimethoxyxanthone, 1-hydroxy-3,5,6,7-tetramethoxyxanthone, 1-hydroxy-3,5,6,7,8-pentamethoxyxanthone, 1-hydroxy-3,7,8-trimethoxyxanthone and 1,8-dihydroxy-3,5,6,7-tetramethoxyxanthone) were isolated and identified by spectroscopic means (nuclear magnetic resonance, mass spectroscopy, and UV). Subsequently, a high-performance liquid chromatography/diode array detection method was developed for the determination of these and other methoxylated xanthones occurring in the chloroform extract of small centaury aerial parts. The methodology developed was applied to twelve samples, and in all of them, nine xanthones were identified and quantified. This methodology can be considered complimentary to the one proposed by the European Pharmacopoeia.     

Podzolization as a deferralitization process: a study of an Acrisol–Podzol sequence derived from Palaeozoic sandstones in the northern upper Amazon Basin

Morphological, geochemical and mineralogical studies were carried out in a representative soil catena of the low‐elevation plateaux of the upper Amazon Basin to interpret the steps and mechanisms involved in the podzolization of low‐activity clay soils. The soils are derived from Palaeozoic sandstones. They consist of Hydromorphic Podzols under tree savannah in the depressions of the plateaux and predominantly of Acrisols covered by evergreen forest elsewhere. Incipient podzolization in the uppermost Acrisols is related to the formation of organic‐rich A and Bhs horizons slightly depleted in fine‐size particles by both mechanical particle transfer and weathering. Weathering of secondary minerals by organic acids and formation of organo‐metallic complexes act simultaneously over short distances. Their vertical transfer is limited. Selective dissolution of aluminous goethite, then gibbsite and finally kaolinite favour the preferential cheluviation of first Fe and secondly Al. The relatively small amount of organo‐metallic complexes produced is related to the quartzitic parent materials, and the predominance of Al over Fe in the spodic horizons is due to the importance of gibbsite in these low‐activity clay soils. Morphologically well‐expressed podzols occur in strongly iron‐depleted topsoils of the depression. Mechanical transfer and weathering of gibbsite and kaolinite by organic acids is enhanced and leads to residual accumulation of sands. Organo‐metallic complexes are translocated in strongly permeable sandy horizons and impregnate at depth the macro‐voids of embedded soil and saprolite materials to form the spodic Bs and 2BCs horizons. Mechanical transfer of black particulate organic compounds devoid of metals has occurred later within the sandy horizons of the podzols. Their vertical transfer has formed well‐differentiated A and Bh horizons. Their lateral removal by groundwater favours the development of an albic E horizon. In an open and waterlogged environment, the general trend is therefore towards the removal of all the metals that have initially accumulated as a response to the ferralitization process and have temporarily been sequestrated in organic complexes in previous stages of soil podzolization.     

Flow cytometric assay in peripheral blood of dogs--reference values for leukocytes from Brazilian beagles

Use of domestic reference values in the flow cytometry analysis is known to improve its accuracy by integrating local variations as gender, race and age. Up to date application of flow cytometry in veterinary medicine has been limited to describe the percentual values just for peripheral lymphocytes subsets of blood. We now report establishment of reference values for a wide range of proportional and absolute numbers of peripheral blood leukocytes, including T cells subsets, B cells, monocytes and eosinophils, applicable to the healthy population of Beagles in Brazil and other regions with similar demographic characteristics. Normal reference values were also established to estimate the gender-related differences. This information will provide clinical aid in the evaluation of immunologic status as well as standard values for experimental animals of dogs from Brazil and other similar regions.     

Gene discovery and expression profiling in porcine Peyer's patch

Peyer's patches of the intestinal mucosa are essential for host defense and immune regulation in the enteric system. To better understand molecular mechanisms of Peyer's patch function, we have screened for differentially expressed genes specific to Peyer's patch. cDNA libraries were created from normal Peyer's patch, immune stimulated Peyer's patch, and pooled cDNA subtracted with fibroblast RNA. From the subtracted library, 3687 expressed sequence tags (ESTs), representing 2414 unique nucleotide sequences, were isolated, identified by BLAST searches against public databases, and spotted onto a microarray for gene expression profiling. Approximately 30% of these ESTs BLAST to genes of unknown function and 20% have no known homology in the public databases (novel genes). Of the novel genes, 70% are expressed in normal immune tissues by microarray analysis, suggesting that at least 371 of the unidentified EST sequences from the subtracted library are novel porcine genes and can now be further characterized to determine their function in the porcine Peyer's patch. We surmise that the products of these genes participate in biochemical and cellular functions related to the unique immunological and gastroenterological functions of the small intestine. The BLAST and gene ontology information for each of the subtracted library EST sequences, the normal and immune stimulated libraries, and the microarray are all valuable resources that will facilitate further examination of the biological function of porcine Peyer's patch tissue.     

Mitochondrial damage as an early event of monensin-induced cell injury in cultured fibroblasts L929

The present study was designed to identify, submicroscopically, the primary organelle or target structure for monensin in cultured murine fibroblasts L929. In addition, the effect of the drug on cell size and surface membranes of the cells were analysed; cellular proliferation, collagen secretion, and necrosis and apoptosis were re-evaluated. At the lowest concentration of monensin the foremost ultrastructural alteration occurred in the mitochondria, characterized by increased matrix density with disorganized and less distinct crystae. Incubation with monensin at higher concentrations resulted in severe mitochondrial damage and marked dilatation of the Golgi apparatus and rough endoplasmic reticulum cisternae. Fibroblasts exposed to higher concentrations of monensin were enlarged with decreased number of filopodia and hollows in the surface membrane. Moreover, monensin inhibited the cell proliferation, increased immunohistochemical positiveness for collagen type I in a dose-dependent manner, and, at high concentrations, caused cell necrosis whereas apoptosis was not induced. Taken together, these results show that monensin induces early mitochondrial damage, possibly causing an energy deficit that led to inhibition of fibroblasts proliferation and accumulation of collagen causing dilatation of Golgi apparatus and rough endoplasmic reticulum. Moreover, the mitochondrial damage would also explain the monensin-induced necrosis.     

Influence of murine Toxocara canis infection on plasma and bronchoalveolar lavage fluid eosinophil numbers and its correlation with cytokine levels

Toxocara canis is a nematode of the Ascaridae family that normally parasites the small intestine of canid species. Humans are accidentally infected upon ingestion of embryonated eggs, and can manifest several clinical alterations such as fever, hepatomegaly, splenomegaly, respiratory symptoms, muscle pain and anorexia. In the present work, we investigated the kinetics of tissue distribution of L2 larva in lungs, liver, kidney, brain, skeletal muscle and myocardium. Also, we analyzed the blood and bronchoalveolar lavage fluid (BAL) for levels of IL-6, IFN-gamma, eotaxin and Regulated on Activation Normal T Cell Expressed and Secreted (RANTES) in experimental murine T. canis infection. We observed liver, lung and kidney lesions correlated to larva migration as early as the first day of infection. After the seventh post-infection day, larva could also be detected in brain, skeletal muscle and heart, as an indicator of biphasic migration pattern. Increased inflammatory activity was detected in BAL and plasma of infected animals, as was an intense eosinophil migration associated with an increase in the levels of all the cytokines studied. In conclusion, our results establish a tight correlation between tissue lesions caused by larva migration and increased plasma levels of pro-inflammatory and eosinophil chemotactic cytokines. Thus, murine T. canis infection may prove to be useful in understanding the role of cytokines in infection.