Comparação de dois métodos de extração de Xanthomonas campestris pv. phaseoli em sementes de feijoeiro

The objective of this research was to evaluate the effectiveness of the extraction of Xanthomonas campestris pv. phaseoli from naturally infected dry bean seeds. Extraction methods tested included soaking whole seeds in sterilized saline phosphate buffer and crushing seeds after soaking in sterilized saline phosphate buffer. The bacterium was isolated on a semiselective agar medium. The seed crushing method was found to be more effective. The bacterium strains isolated were pathogenic to bean leaves, reacted with X. campestris pv. phaseoli antiserum, and had morphological and physiological/biochemical characteristics typical of the X. campestris pv. phaseoli.

Occurrence, distribution, and relative incidence of five viruses infecting cucurbits in the state of Sao Paulo, Brazil

Cucurbits species grown in 38 of 40 agricultural regions in the state of Sao Paulo, Brazil, were surveyed for the relative incidence of Cucumber mosaic virus (CMV), Papaya ringspot virus-type W (PRSV-W), Watermelon mosaic virus-2 (WMV-2), Zucchini lethal chlorosis virus (ZLCV), and Zucchini yellow mosaic virus (ZYMV) during May 1997 and June 1999. Samples from 621 plants, representing eight cultivated species, six wild species, and one commercial hybrid (Cucurbita moschata x C. maxima), were analyzed by plate trapped antigen enzyme-linked immunosorbent assay (PTA-ELISA). PRSV-W and ZYMV were the most frequently found viruses, accounting for 49.1 and 24.8%, respectively, of 605 samples tested. ZLCV, CMV, and WMV-2 were detected in 7.8, 6.0, and 4.5% of 612, 497, and 423 samples tested, respectively. Double infection was found in 97 samples, and triple infection was found in 10 samples. Quadruple infection was detected in one C. pepo sample. Plants that were symptomatic but negative by PTA-ELISA might be due to abiotic agents, infection by virus for which antiserum was not available, such as Squash mosaic virus, or infection with an as yet uncharacterized virus.

A novel biological activity for galectin-1: Inhibition of leukocyte-endothelial cell interactions in experimental inflammation

Galectin-1 (Gal-1), the prototype of a family of β -galactoside-binding proteins, has been shown to attenuate experimental acute and chronic inflammation. In view of the fact that endothelial cells (ECs), but not human polymorphonuclear leukocytes (PMNs), expressed Gal-1 we tested here the hypothesis that the protein could modulate leukocyte-EC interaction in inflammatory settings. In vitro, human recombinant (hr) Gal-1 inhibited PMN chemotaxis and trans-endothelial migration. These actions were specific as they were absent if Gal-1 was boiled or blocked by neutralizing antiserum. In vivo, hrGal-1 (optimum effect at 0.3 μg equivalent to 20 pmol) inhibited interleukin-1β-induced PMN recruitment into the mouse peritoneal cavity. Intravital microscopy analysis showed that leukocyte flux, but not their rolling velocity, was decreased by an anti-inflammatory dose of hrGal-1. Binding of biotinylated Gal-1 to resting and post-adherent human PMNs occurred at concentrations inhibitory in the chemotaxis and transmigration assays. In addition, the pattern of Gal-1 binding was differentially modulated by PMN or EC activation. In conclusion, these data suggest the existence of a previously unrecognized function of Gal-1, that is inhibition of leukocyte rolling and extravasation in experimental inflammation. It is possible that endogenous Gal-1 may be part of a novel anti-inflammatory loop in which the endothelium is the source of the protein and the migrating PMNs the target for its anti-inflammatory action.

Action of anti-bothropic factor isolated from Didelphis marsupialis on renal effects of Bothrops erythromelas venom

Acute renal failure is the most common complication in the lethal cases caused by snakebites in Brazil. Among the Brazilian venom snakes, Bothrops erythromelas is responsible for the majority of accidents in Northeastern Brazil. Didelphis marsupialis serum could inhibit myonecrotic, hemorrhagic, edematogenic hyperalgesic and lethal effects of envenomation determined by ophidian bites. In the present study, we evaluated the action of the anti-bothropic factor isolated from D. marsupialis on the renal effects promoted by B. erythromelas venom without systemic interference. Isolated kidneys from Wistar rats were perfused with Krebs-Henseleit solution containing 6% bovine serum albumin. We analyzed renal perfusion pressure (PP), renal vascular resistance (RVR), glomerular filtration rate (GFR), urinary flow (UF), and the percentages of sodium and potassium tubular transport (%TNa +, %TK +). The B. erythromelas venom (10 μg mL -1) decreased the PP (ct=108.71±5.09 mmHg; BE=65.21±5.6 mmHg*) and RVR (ct=5.76±0.65 mmHg mL -1 g -1 min -1; BE=3.10±0.45 mmHg mL -1 g -1 min -1*) . On the other hand, the GFR decreased at 60 min (ct 60=0.76±0. 07 mL g -1 min -1; BE 60=0.42±0.12 mL g -1 min -1*) and increased at 120 min (ct 120=0.72±0.01 mL g -1 min -1; BE 120=1.24±0.26 mL g -1 min -1*). The UF increased significantly when compared with the control group (ct=0.14±0.01 mL g -1 min -1; BE=0.47±0.08 mL g -1 min -1*). The venom reduced the %TNa + (ct 90=79.18±0.88%; BE 90=58.35±4.86%*) and %TK + (ct 90=67.20±4.04%; BE 90=57. 32±5.26%*) The anti-bothropic factor from D. marsupialis (10 μg mL -1) incubated with B. erythromelas venom (10 μg mL -1) blocked the effects on PP, RVR, %TNa +, and %TK +, but was not able to reverse the effects in UF and GFR promoted by venom alone. However, the highest concentration of D. marsupialis serum (30 μg mL -1) reversed all the renal effects induced by the venom. In conclusion, B. erythromelas venom altered all the renal functional parameters evaluated and the anti-bothropic factor from D. marsupialis was able to inhibit the effects induced by the venom in isolated kidney. © 2005 Elsevier Ltd. All rights reserved.

Laboratory evaluation of young ovines inoculated with natural or 60Co-irradiated Crotalus durissus terrificus venom during hyperimmunization process

Laboratory profile of young ovines was studied in order to evaluate and compare their antiserum production from natural and Cobalt-60 irradiated Crotalus durissus terrificus (C.d.t.) venoms. The parameters analyzed included complete blood count, and urea, creatinine, aspartate aminotransferase, total proteins, albumin and globulin serum measurements. Three groups of six animals each were used. Group 1 (G1) received natural C.d.t. venom; Group 2 (G2) received irradiated C.d.t. venom; and Group 3 (G3) was used as control and did not receive venom, only adjuvants, using seven venom inoculations. During the experimental period, animals were fortnightly weighed. According to clinical and weight evaluation, sheep in post-weaning phase showed no changes in their physiological profiles but had excellent weight gain. The parameters analyzed were not statistically different (p<5%) among the groups tested. The hyperimmunization process was successfully accomplished with the production of specific antibodies against Crotalus durissus terrificus venom. Results bring a new possibility of utilizing ovines in the commercial production of anticrotalic serum, which may be used to treat human and animal envenomation. Its production cost may be reduced by subsequent use of hyperimmunized sheep for human consumption.

Biochemical and hematological study of goats envenomed with natural and 60Co-irradiated bothropic venom

Venoms from snakes of the Bothrops genus are proteolytic, coagulant, hemorrhagic and nephrotoxic, causing edema, necrosis, hemorrhage and intense pain at the bite site, besides systemic alterations. Many adjuvants have been added to the venom used in the sensitization of antiserum-producer animals to increase antigenic induction and reduce the envenomation pathological effects. Gamma radiation from 60Co has been used as an attenuating agent of the venoms toxic properties. The main objective was to study, comparatively, clinical and laboratory aspects of goats inoculated with bothropic (Bothrops jararaca) venom, natural and irradiated from a 60Co source. Twelve goats were divided into two groups of six animals: GINV, inoculated with 0.5mg/kg of natural venom; and GIIV, inoculated with 0.5mg/kg of irradiated venom. Blood samples were collected immediately before and one, two, seven, and thirty days after venom injection. Local lesions were daily evaluated. The following exams were carried out: blood tests; biochemical tests of urea, creatinine, creatine kinase (CK), aspartate amino-transferase (AST) and alanine amino-transferase (ALT); clotting time; platelets count; and total serum immunoglobulin measurement. In the conditions of the present experiment, irradiated venom was less aggressive and more immunogenic than natural venom.

Resposta imune e capacidade de neutralização de anticorpos produzidos em ovinos jovens imunizados com veneno de Crotalus durissus terrificus nativo e irradiado com Cobalto 60

The ELISA technique was used to evaluate and compare young ovine humoral immune response during crotalic antiserum production. Animals were clinically evaluated throughout this process, and the neutralizing capacity of antisera raised against natural (NV) and Cobalt-60 irradiated (IrV) venoms of Crotalus durissus terrificus (C.d.t.) was verified by means of in vitro challenges. Three groups of six animals each were used: G1 received NV; G2 was inoculated with IrV; and G3 was used as control. Animals received six immunizations during 84 days at 14-day intervals. ELISA of antibody profile showed significant difference (p<5%) between experimental groups (G1antiserum production. The neutralizing capacity of antiserum raised against IrV was fivefold higher than that of antiserum raised against NV. Results showed a new possibility of using ovines to produce commercial crotalic antiserum, which may be employed in the treatment of human and animal envenomation. Production cost might be reduced by the subsequent utilization of hyperimmunized ovines as food.

Caracterização molecular parcial e localização subcelular de um isolado brasileiro do vírus do mosaico do feijoeiro do sul dos E.U.A

Pós-graduação em Genética - IBILCE

Caracterização da variabilidade genética em isolados de Xanthomonas albilineans oriundos de diferentes regiões do Brasil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Proteína capsidial do Rupestris stem pitting-associated vírus: seqüenciamento do gene, expressão em Escherichia coli, purificação e produção de anti-soro policlonal

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Variabilidade de potyvirus infectando Capsicum spp. no estado de São Paulo

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ocorrência de viroses em mandioquinha-salsa (Arracacia xanthorrhiza brancroft) nas principais regiões produtoras do Brasil

Pós-graduação em Agronomia (Proteção de Plantas) - FCA

Identificação e purificação de isolados de Cucumber mosaic virus e triagem de genótipos de pimentão para resistência

Pós-graduação em Agronomia (Proteção de Plantas) - FCA

Caracterização biofísica e produção de antissoro policlonal contra a capa proteica recombinante do Southem bean mosaic virus

Pós-graduação em Genética - IBILCE