Biological Institute of Sao Paulo

São Paulo, Brazil

Biological Institute of Sao Paulo

São Paulo, Brazil

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Gotardo A.T.,University of Sao Paulo | Haraguchi M.,Biological Institute of Sao Paulo | Raspantini P.C.F.,University of Sao Paulo | Dagli M.L.Z.,University of Sao Paulo | Gorniak S.L.,University of Sao Paulo
Avian Pathology | Year: 2017

Senna occidentalis is a toxic leguminous plant found in many tropical and subtropical regions of the world and causes poisoning mainly in confined animals. The seeds are the most toxic part of the plant and may be present in animal rations. The main toxic component of the S. occidentalis seed is a dianthrone, an anthraquinone-derived compound that affects mitochondrial function. This study evaluated the effects on egg production of low-level contamination of the S. occidentalis in the laying hens’ diet. Forty-eight one-day-old pullets were randomly allocated into two treatment groups: control, birds that received no experimental treatment; and external and internal tegument (ET/IT), birds that received a diet containing 0.2% of ET/IT of S. occidentalis seeds throughout their life cycle (42 weeks). The birds were monitored for clinical signs of poisoning, and the production and quality of eggs were recorded. Necropsies were conducted at the end of the experimental period. None of the layers showed any clinical signs of poisoning, decreases in feed intake or alterations of the body weight gain. A marked reduction in egg production and, consequently, a lower feed efficiency in ET/IT group were measured. Ovaries were the most affected organ in birds from the ET/IT group, and yolk leaking and dysplasia of the inner layer of the vitelline membrane were observed. S. occidentalis was shown to be toxic for laying hens. Considering these results, it is feasible to assume that the constant presence of low concentrations of S. occidentalis seeds in rations represents a threat to the poultry industry. © 2017 Houghton Trust Ltd


PubMed | Biological Institute of Sao Paulo and University of Sao Paulo
Type: | Journal: Evidence-based complementary and alternative medicine : eCAM | Year: 2015

Hebanthe paniculata roots (formerly Pfaffia paniculata and popularly known as Brazilian ginseng) show antineoplastic, chemopreventive, and antiproliferative properties. Functional properties of these roots and their extracts are usually attributed to the pfaffosidic fraction, which is composed mainly by pfaffosides A-F. However, the therapeutic potential of this fraction in cancer cells is not yet entirely understood. This study aimed to analyze the antitumoral effects of the purified pfaffosidic fraction or saponinic fraction on the human hepatocellular carcinoma HepG2 cell line. Cellular viability, proliferation, and apoptosis were evaluated, respectively, by MTT assay, BrdU incorporation, activated caspase-3 immunocytochemistry, and DNA fragmentation assay. Cell cycle was analyzed by flow cytometry and the cell cycle-related proteins were analyzed by quantitative PCR and Western blot. The cells exposed to pfaffosidic fraction had reduced viability and cellular growth, induced G2/M at 48h or S at 72h arrest, and increased sub-G1 cell population via cyclin E downregulation, p27(KIP1) overexpression, and caspase-3-induced apoptosis, without affecting the DNA integrity. Antitumoral effects of pfaffosidic fraction from H. paniculata in HepG2 cells originated by multimechanisms of action might be associated with cell cycle arrest in the S phase, by CDK2 and cyclin E downregulation and p27(KIP1) overexpression, besides induction of apoptosis through caspase-3 activation.


Catroxo M.H.B.,Biological Institute of Sao Paulo | Martins A.M.C.R.P.F.,Biological Institute of Sao Paulo | Petrella S.,Adolfo Lutz Institute | Curi N.A.,Biological Institute of Sao Paulo | Melo N.A.,Biological Institute of Sao Paulo
International Journal of Morphology | Year: 2011

The pigeon feces are vehicle of diseases both for humans and other animal species. In these birds, the most important viral diseases of the digestive tract are transmitted by the paramyxovirus, adenovirus and coronavirus. Avian paramyxoviruses have been isolated from a variety of species of free living and domestic birds worldwide, with several symptoms and clinical signs and economic losses. Paramyxoviruses belong to the Paramyxoviridae family and Avulovirus genus that includes nine serotypes (APMV 1 to 9). Avian adenoviruses belong to the Adenoviridae family and Aviadenovirus genus. In pigeons, cause classical adenovirosis and necrotizing hepatitis. The respiratory and enteric tracts are common targets of coronavirus. They belong to the Coronaviridae family and to 3a and 3c groups. In this study, we described the presence of viral agents in free-living pigeon feces (Columba livia) from the city of São Paulo, Brazil. The feces were processed by negative staining technique (rapid preparation) for transmission electron microscopy. In this technique paramyxoviruses particles, pleomorphic, roughly spherical or filamentous, measuring 100 to 500 nm of diameter containing an envelope covered by spikes, with characteristic helical herring-bone-like nucleocapsid, measuring 15 to 20 nm in diameter, were visualized in 45 (79%) out of 57 feces samples. In 2 (3.5%) samples, paramyxovirus and adenovirus particles were simultaneously visualized. Adenovirus particles were isometric, spherical, characterized as "complete" or "empty", measuring between 70 and 90 nm in diameter. Paramyxovirus and coronavirus particles were detected in 3 (5.2%) samples. Coronaviruses were pleomorphic with a diameter of 75-160 nm containing a solar corona-shaped envelope, with projections of approximately 20 nm of diameter. Seven (12.3%) samples were negative for viral particles.


Catroxo M.H.B.,Biological Institute of Sao Paulo | Martins A.M.C.R.P.F.,Biological Institute of Sao Paulo | Petrella S.,Adolfo Lutz Institute | Souza F.,Biological Institute of Sao Paulo | Nastari B.D.B.,Biological Institute of Sao Paulo
International Journal of Morphology | Year: 2013

Papillomaviruses, belonging to the Papillomaviridae family, are small oncogenic viruses, causing papillomas and fibropapillomas in the mucosal and cutaneous epithelia of several animals. In bovine species, thirteen types (BPV 1-13) were characterized to date. In this study, the occurrence of papillomatosis in four outbreaks in cattle herds, coming from Brazilian states were registered. The papillomatous lesions were found located in the teats, udders, head and neck. Under the transmission electron microscope, by the negative staining technique, it was possible to visualized rounded-format papillomavirus, with icosahedral symmetry, characterized as "full" and "empty" particles, measuring on average 60 nm in diameter, in all the 40 samples observed of skin lesion fragments. The ultrathin sections revealed the presence of groups of viral, intranuclear, rounded particles measuring 35 nm in diameter and tubular particles with a diameter of 35-39 nm. At immunoelectron microscopy technique, positivity obtained was marked by the presence of aggregates of viral particles formed by the antigen-antibody interaction. In the immunocytochemistry technique, the antigen-antibody reaction showed colloidal gold particles evenly distributed over the surface of the virus. These results showed the importance of the transmission electron microscopy techniques in the diagnosis of bovine papillomatosis that can be used in routine procedures to identify viral agent of this important disease.


Latorre A.O.,University of Sao Paulo | Caniceiro B.D.,University of Sao Paulo | Wysocki H.L.,Biological Institute of Sao Paulo | Haraguchi M.,Biological Institute of Sao Paulo | And 2 more authors.
Food and Chemical Toxicology | Year: 2011

We have previously shown that bracken fern (Pteridium aquilinum) has immunomodulatory effects on mouse natural killer (NK) cells by reducing cytotoxicity. Alternatively, it has been demonstrated that selenium can enhance NK cell activity. Therefore, the aims of the present study were to evaluate if ptaquiloside, the main toxic component found in P. aquilinum, is responsible for the immunotoxic effects observed in mice, and if selenium supplementation could prevent or even reverse these effects. Male C57BL/6 mice were administered the P. aquilinum extract by daily gavage for 30. days, and histological analyses revealed a significant reduction in splenic white pulp area that was fully reversed by selenium treatment. In addition, mice administered ptaquiloside by daily gavage for 14. days demonstrated the same reduction of NK cell activity as the P. aquilinum extract, and this reduction was prevented by selenium co-administration. Lastly, non-adherent splenic cells treated in vitro with an RPMI extract of P. aquilinum also showed diminished NK cell activity that was not only prevented by selenium co-treatment but also fully reversed by selenium post-treatment. The results of this study clearly show that the immunosuppressive effects of P. aquilinum are induced by ptaquiloside and that selenium supplementation can prevent as well as reverse these effects. © 2010 Elsevier Ltd.


Caniceiro B.D.,University of Sao Paulo | Latorre A.O.,University of Sao Paulo | Fukumasu H.,University of Sao Paulo | Sanches D.S.,University of Sao Paulo | And 2 more authors.
Journal of Immunotoxicology | Year: 2015

Pteridium aquilinum (bracken fern), one of the most important toxic plants in the world, contains the toxic norsequiterpene ptaquiloside that induces cancers in humans and farm animals. Previous studies in the laboratory demonstrated immunotoxic effects produced by ptaquiloside, which are characterized by suppression of natural killer (NK) cell activity (i.e. cytotoxicity and interferon [IFN]-γ production). However, it is unknown whether these immunosuppressive effects could contribute to carcinogenesis in situ in general because of the important function of NK cells in innate killing of tumor cells. This study assessed the impact of P. aquilinum-induced immunosuppression on urethane-induced lung cancer in C57BL/6 mice. Adult mice were treated with an extract of P. aquilinum (30 g/kg/day) by gavage once daily for 14 days, followed by gavage (5 days/week) during an 11-week period that was accompanied by treatment with urethane (1 g/kg) via once-weekly intraperitoneal injection; 20 weeks after the end of the treatment period, all lungs were evaluated. The results indicated there was a significant increase in lung nodule number as well as in multiplicity of lesions in mice treated with both P. aquilinum and urethane (PU group) compared to values in mice treated only with the urethane (U group). In addition, histologic evaluation revealed a 76% increase in the rate of lung adenomas and a 41% increase in rate of bronchiolization of alveoli in the mice from the PU group compared to levels seen in mice within the U group. Taken together, the results here show for the first time that immunosuppressive effects of P. aquilinum could increase the risk of cancer formation in exposed hosts. © 2015 Informa Healthcare USA, Inc.


Elias F.,University of Sao Paulo | Latorre A.O.,University of Sao Paulo | Pipole F.,University of Sao Paulo | Haraguchi M.,Biological Institute of Sao Paulo | And 2 more authors.
Food and Chemical Toxicology | Year: 2011

This study is the first in the literature to focus attention on the possible immunotoxic effect of integerrimine N-oxide content in the butanolic residue (BR) of Senecio brasiliensis, a poisonous hepatotoxic plant that contains pyrrolizidine alkaloids (PAs). PAs have been reported as a pasture and food contaminant and as herbal medicine used worldwide and are responsible for poisoning events in livestock and human beings. After the plant extraction, BR extracted from Senecio brasiliensis was found to contain approximately 70% integerrimine N-oxide by elemental and spectral analyses ( 1H and 13C NMR), which was administered to adult male Wistar Hannover rats at doses of 3, 6 and 9mg/kg for 28days. Body weight gain, food consumption, lymphoid organs, neutrophil analysis, humoural immune response, cellular immune response and lymphocyte analysis were evaluated. Our study showed that integerrimine N-oxide could promote an impairment in the body weight gain, interference with blood cell counts and a reducing T cell proliferative activity in rats; however, no differences in the neutrophil activities, lymphocytes phenotyping and humoural and cellular immune responses were observed. It is concluded that doses of integerrimine N-oxide here employed did not produce marked immunotoxic effects. © 2011 Elsevier Ltd.


dos Santos R.F.,Federal University of Santa Maria | Blume E.,Federal University of Santa Maria | Muniz M.F.B.,Federal University of Santa Maria | Heckler L.I.,Federal University of Santa Maria | And 4 more authors.
Plant Disease | Year: 2014

Cultivated grapevine (Vitis labrusca and V. vinifera) is of considerable economic importance to the Brazilian fruit industry for both fresh market consumption and for the production of wines, sparkling beverages, and juices. Black foot disease is caused by fungi of the genera Ilyonectria P. Chaverri & C. Salgado (anamorph: Cylindrocarpon Wollew.), Campylocarpon Halleen, Schroers & Crous, and Cylindrocladiella Boesew. In 2012, 4- to 40-year-old grapevines (Vitis spp.) showing reduced vigor, vascular lesions, necrotic root lesions, delayed budding, vine decline, and death were collected from seven locations at Rio Grande do Sul state, Brazil. Fungal isolations were made from root fragments and crown lesions (at least 2 cm above the bottom) on potato dextrose agar (PDA) medium added with 0.5 g L-1 streptomycin sulfate. Eight isolates were obtained and identified on the basis of morphological features and multi-gene analysis (rDNA-ITS, β-tubulin, and histone H3) as Ilyonectria macrodidyma (Halleen, Schroers & Crous) P. Chaverri & C. Salgado. One representative isolate (Cy5UFSM) was used for more detailed morphological and molecular characterization, and pathogenicity confirmation. When incubated in the dark at 20°C for 7 to 10 days, colonies of felty straw-colored mycelium (3) 4.79 cm diameter on average were observed. No sporodochia or other fruiting bodies were produced on carnation leaf agar (CLA) medium after 30 days. Microconidia that were produced after 5 weeks on spezieller nährstoffarmer agar (SNA) medium with addition of two pieces of 1 cm2 filter paper showed ovoid and ellipsoid shape (6.4 × 3.6 μm) and one-septate macroconidia (17.3 × 4.1 μm). To confirm the species, primer pairs ITS1 and ITS4 (4); Bt2a and Bt2b; and H3-1a and H3-1b (2) were used to amplify the ITS1-5.8S rRNA-ITS2, part of the β-tubulin and histone H3 genes, respectively. Sequences of these three regions showed 99, 100, and 100% of homology with I. macrodidyma, respectively. To confirm pathogenicity, 4-month-old rooted cuttings of V. labrusca cv. Bordô were inoculated by immersing them in a conidial suspension of the isolate (106 conidia ml-1) for 60 min (1). Thirty days later, inoculation was performed again by drenching the crown with 40 ml of 106 conidia ml-1 suspension to ensure infection of the roots. In the control treatment, plants were inoculated with sterile distilled water. Plants inoculated with I. macrodidyma showed necrosis of the leaf ribs, reduction in root mass, root and crown necrosis, browning of vessels, drying of shoots, and death. I. macrodidyma was re-isolated from the crown necrosis and vascular lesions, confirming Koch's postulates. To our knowledge, this is the first report of I. macrodidyma associated with black foot disease of grapevine in Brazil, which poses considerable threat to the industry unless management options are realized. © The American Phytopathological Society.


Martins A.M.C.R.P.F.,Biological Institute of Sao Paulo | Bersano J.G.,Biological Institute of Sao Paulo | Ogata R.,Biological Institute of Sao Paulo | Amante G.,Biological Institute of Sao Paulo | And 2 more authors.
International Journal of Morphology | Year: 2013

The porcine transmissible gastroenteritis is a highly severe contagious disease, caused by virus of the Coronaviridae family, genus Coronavirus. Its epizootic shape can reach a rate of up to 100% mortality in piglets under two weeks of age as a result of severe dehydration. In this study fragments of small intestine and stool samples were collected from 75 autopsied pigs from properties. The samples of the fragments were frozen and sent to the Laboratory of Electron Microscopy, Instituto Biológico, SP, Brazil, for histological and transmission electron microscopic analyses. According to histological H&E technique, atrophy, villous necrosis and destruction of the enterocytes were observed in 35 (46.6%) out of the 75 fragments of the small intestine samples. On the immunohistochemistry technique 19 (25.3%) fragments were positively stained with DAB in the Ag-Ac reaction (MabTGEV). In 19 (25.3%) positive samples analyzed by in situ hybridization, a brown stain of enterocytes was observed, mainly in the epithelial cells of the villi. By the negative staining technique, we visualized enveloped, pleomorphic coronavirus particles, with typical radial projections resembling solar corona, with 140 nm diameter in 21 samples (28%) of the small intestine fragments and in 16 (21.3%) stool samples. In the ultrathin sections of 21 (28%) samples of small intestine, complete coronavirus particles with 80 nm diameter were seen among the microvilli and in the cytoplasm of epithelial cells. Immature particles with 60 nm diameter, budding from cell membrane and from a rough endoplasmic reticulum and also inside the vacuoles were visualized. In 19 (25.3%) out of 75 analyzed samples of small intestine, the antigenantibody interaction was characterized by aggregation of viral particles in the immunoelectron microscopy technique. In the immunocytochemistry technique, the antigen-antibody interaction was strongly enhanced by the dense colloidal gold particles over the coronavirus in 19 (25.3%) out of the 75 samples of fragments of small intestine examined.


Lazarotto M.,Federal University of Santa Maria | Muniz M.F.B.,Federal University of Santa Maria | Poletto T.,Federal University of Santa Maria | Dutra C.B.,Federal University of Santa Maria | And 3 more authors.
Plant Disease | Year: 2012

Conspicuous leaf spots in combination with fruit spots were observed for the first time in April and May 2010 on a 30-ha pecan [Carya illinoensis (Wangenh.) K. Koch] orchard in the state of Rio Grande do Sul, Brazil. Initially, tiny grey spots were observed on leaves and, over time, the spots expanded to become gray to light brown circles surrounded by a dark brown border, followed by leaves falling. Eventually, fruits were also attacked, with typical symptoms beginning with tiny water soaked spots which then became necrotic. The disease was also observed in pecan nursery and field seedlings. Isolation of the pathogen from symptomatic leaves and morphological identification by conidia characters (number of cells, color, hyaline terminal cells, number of appendages) revealed Pestalotiopsis sp. (2) as the causal agent of the disease. Conidia constituted of transverse septa with four dark intermediate sections and two hyaline terminal sections. One of the terminal sections presented two or three apical appendages. Conidia averaged 6.88 μm wide × 31.00 μm long, not considering the apical appendages. Primers ITS 1 and ITS 4 were used to amplify the internal transcribes spacer ITS 1-5.8S-ITS 2 region. Nucleotide sequences were 99% similar to Pestalotiopsis clavispora (G.F. Atk.) Steyaert. Conidia produced on potato dextrose agar medium were used to inoculate 8 plants with a spore suspension of 2.0 × 106 conidia/ml. Eight additional plants were used as control (non-inoculated). The inoculation was performed by spraying the suspension onto the leaves of Pecan seedlings and the plants were incubated for 72 h in a humid chamber (1). All inoculated plants showed symptoms 25 days after inoculation and the fungus was reisolated. The pathogenicity test was repeated once. Ten more isolates collected from four different cities in the same state were identified as Pestalotiopsis spp. by morphological characterization and pathogenicity was confirmed. Because this disease causes losses on production of nuts indirectly by reducing photosynthetically active area when the pathogen attacks leaves and directly when attacking fruits, it may restrict the production where the pathogen occurs. On some orchards in the state of Rio Grande do Sul, the attack rate reached 80% of the plants. P. clavispora has been reported causing stem end-rot of avocado in Chile (3), but this note constitutes the first report, to our knowledge, of P. clavispora causing leaf spot on C. illinoensis in Brazil. © The American Phytopathological Society.

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