Diagnosis of canine brucellosis: Comparison between serological and microbiological tests and a PCR based on primers to 16S-23S rDNA interspacer

A pair of primers directed to 16S-23S rDNA interspacer (ITS) was designed directed to Brucella genetic sequences in order to develop a polymerase chain reaction (PCR) putatively capable of amplifying DNA from any Brucella species. Nucleic acid extracts from whole-blood from naive dogs were spiked with decreasing amounts of Brucella canis RM6/66 DNA and the resulting solutions were tested by PCR. In addition, the ability of PCR to amplify Brucella spp. genetic sequences from naturally infected dogs was evaluated using 210 whole-blood samples of dogs from 19 kennels. The whole-blood samples collected were subjected to blood culture and PCR. Serodiagnosis was performed using the rapid slide agglutination test with and without 2-mercaptoethanol. The DNA from whole blood was extracted using proteinase-K, sodium dodecyl sulphate and cetyl trimethyl ammonium bromide followed by phenol-chloroform purification. The PCR was capable of detecting as little as 3.8 fg of Brucella DNA mixed with 450 ng of host DNA. Theoretically, 3.8 fg of Brucella DNA represents the total genomic mass of fewer than two bacterial cells. The PCR diagnostic sensitivity and specificity were 100%. From the results observed in the present study, we conclude that PCR could be used as confirmatory test for diagnosis of B. canis infection.

The Brazilian pediatric myelodysplastic cooperative group strategies: are they relevant to improve educational approach and correct diagnosis?

Brazil is a wide country with huge contrasts. Its peculiarities can highlight environmental factors that could influence the frequencies of different cancers. The standard treatment and results achieved from several different areas of the country may not be found in others. The establishment of a national cooperative group has the potential to improve outcomes. The The Brazilian Cooperative Group on Pediatric Patients with Myelodysplastic Syndrome (BCG-MDS-PED) was first organized in January 1997 as a working group of hematologists, pediatric oncologists, pediatric-hematologists, molecular biologists and other professionals in order to study pediatric (age < 18 years) MDS. Six distinct subcommittees constituted with members from several universities: cytology, histopathology, clinical, cytogenetics, molecular biology and epidemiology. The goals of the BCG-MDS-PED were: (i) to offer support for diagnosis and orientation for treatment; (ii) educational Support for the colleagues all over the country and (iii) research on pathogenesis and new approaches for pediatric MDS patients. There are socio-economical differences among the five regions of the country. The BCG-MDS-PED believes that it is absolutely necessary to Study the clinical, cellular, molecular and epidemiological aspects of MDS, taking in account these peculiar differences among populations and regions. Since 1997, 114 pediatric cases were referred to the BCG-MDS-PED from 21 centres. Seven Brazilian states have sent cases to the group, 31 patients were referred from universities, 73 patients from pediatric oncology units (foundations) and 10 patients came from private clinics. Some of these patients have been followed up and/or treated by the physician who referred them to the BCG-MDS-PED for confirmation of the initial diagnosis. The majority of these physicians have required orientation on diagnostic and treatment issues, as well as to complete cytogenetic and molecular studies. From these 114 patients, 64 patients were confirmed as MDS. We believe that, the more numerous the MDS-studied cases, the more experienced will be the referee group on clinical and laboratory features on childhood MDS in Brazil. (C) 2002 Published by Elsevier B.V. Ltd.

Effect of doses of fungicides and plant resistance activators on the control of Rhizoctonia foliar blight of soybean, and on Rhizoctonia solani AG1-IA in vitro development

Rhizoctonia foliar blight (RFB) of soybean [Glycine max (L.) Merrill] occurs in many tropical and subtropical regions, causing yield reductions of up to 70% and in Brazil, up to 60%. The disease is caused by Rhizoetonia solani AG1-IA and AG1-IB, and by AG2-3 in Japan. RFB occurs in the North, Northeast and Mid-west regions of Brazil. Chemical control remains the only effective method of controlling RFB, but its efficiency depends upon environmental conditions. In this study, 18 fungicides, salicylic acid (SA) and acibenzolar-s-methyl (ASM) were evaluated on R. solani AG1-IA in vitro, by mycelial growth rating and estimating effective concentration for 50% (EC 50) and 90% (EC 90) inhibition of mycelial growth, and in vivo by reduction of disease severity on soybean plants in greenhouse conditions. Mycelial growth was strongly inhibited by the fungicides pyraclostrobin + boscalid and fludioxonil. Preventive fungicide applications were the most effective. Strobilurins were more efficient both in preventive and curative applications. Best results with plant resistance activators were obtained with SA (2.5 mM) sprayed at 20 d before inoculation and with ASM (12.5 mg a.i. l(-1)) 10 d before inoculation. (c) 2005 Elsevier Ltd. All rights reserved.

Adubação foliar. I. Épocas de aplicação de fósforo na cultura da soja

The search for new alternatives in order to increase soybeans productivity has been constant objective of researchers and farmers. The crop responses to phosphorus application in the soil are well defined, being this nutrient very important on its development and yield. The leaf fertilization on this crop appears as a new rationale option, mainly when the plant nutrient levels are low. So, this work aimed to study the effect of phosphorus leaf fertilization, applied at different plant stage, including: V5, R1, R4, V5 + R1, V5 + R4, R1 + R4, V5 + R1 + R4, V5 + R1 + R4 + R6 and test plot. The experiment was installed in a soybeans crop, Monarca cultivar, at Palmital Farm, Ijaci county, Minas Gerais state, Brazil, using a totally randomized design, with 9 treatments and 3 replications. The chelate Quimifol P30 in liquid form with 30% of the nutrient soluble in CNA + water in the, with doses of 21. ha(-1), was utilized as phosphorus source, using the applications performed with a constant pressure CO(2)-nebulizer. The different epochs of phosphorous application significantly altered the grains yield, proportioning significant increases, up to 16% for the V5, V5 + R1, V5 + R4, V5 + R1 + R4, V5 + R1 + R4 + R6 epochs, when compared to the test plot, clearly expressing the positive effect of these applications at V5 stage. The plant height, first legume insertion, and lodging index characteristics were not significantly altered by the different epochs evaluated. It was observed significant response for the nutrient leaf amounts only in the case of K and Zn indices, exclusively in the V5 + R4, and in the V5, V5 + R1 and V5 + R1 + R4 + R6 treatments, respectively.

Aplicação de nitrogênio no solo e pulverização foliar com ácido giberélico e uréia em Setaria anceps Stapf cv. Kazungula

O objetivo do experimento foi verificar o efeito do nitrogénio no solo, parcelado de diferentes formas e aplicado durante o período de "verão", combinado com pulverização foliar de ureia e/ou ácido giberélico, sobre a produção e qualidade de Setaria anceps Stapf cv. Kazungula. O nitrogênio (60 kg/ha/ano) foi aplicado segundo quatro formas de parcelamento: Testemunha (sem nitrogênio), N20M40 (20 kg/ha de N em novembro + 40 kg/ha de N em março), N30M30 (30 kg/ha de N em novembro + 30 kg/ha de N em março), F20M40 (20 kg/ha de N em fevereiro + 40 kg/ha de N em março). No início do período de "inverno" foram aplicados os quatro tipos de pulverizações foliares: Testemunha (sem ácido giberélico ou uréia), uréia a 2%, ácido giberélico a 50 ppm e uréia a 2% + ácido giberélico a 50 ppm. Os resultados revelaram que o parcelamento do nitrogénio no solo não influiu no aumento da produção de matéria seca do capim setária no período de "verão", apesar de proporcionar um incremento da mesma no corte de "inverno". Observou-se, ainda, uma boa distribuição da produção de matéria seca entre "verão" e "inverno", apresentando essa última estação 34,52% da produção total anual, em média. Os diferentes tipos de pulverização não influíram significativamente sobre a produção de matéria seca e a composição química bromatológica da forrageira no período de "inverno".

Application of synthetic peptides to the diagnosis of neurocysticercosis

We tested the possible diagnostic utility of five Taenia saginata oncosphere-derived synthetic peptides in T. solium neurocysticercosis (NC). The five peptides correspond to protein sequences with high antigenic indexes that were cloned from a T. saginata oncosphere cDNA library. The test samples consisted of cerebrospinal fluid (CSF) samples randomly collected from patients referred from Mexican and Brazilian neurological institutes. Indirect enzyme-linked immunosorbent assays (ELISA) were carried out with the peptides either unconjugated or coupled to carrier proteins, and were compared with results obtained using T. solium cyst fluid as a positive control. For active inflammatory NC, the higher sensibility (93%) and specificity (85%) was obtained with peptides HP6-2 and Ts45W-1, respectively, coupled to ovalbumin, in both Mexican and Brazilian patients. Examining the results of the individual peptide assays in combination, in some instances, improved the sensitivity to 100%.

Thermography and ultrasonography in back pain diagnosis of equine athletes

The object of the current study was to evaluate the efficacy of thermography and ultrasonography in the diagnosis of thoracolumbar lesions in Quarter Horse athletes and associate the different types of lesions found with the athletic modality practiced. Twenty-four horses were admitted to the Surgery Service for Large Animals of the Veterinary and Animal Science Faculty, UNESP, Botucatu, Brazil, with complaints of back problems. All the horses were submitted for physical examinations to confirm the existence of thoracolumbar alterations and then for thermography and ultrasonography Thermography was used to map the lesioned areas of this region and ultrasonography for lesion characterization. The lesions found were supraspinous desmitis, interspinous desmitis, dorsal intervertebral osteoarthritis, and impingement of the spinous processes or kissing spines. The existence of a relation between the type of event practiced by the horse and the type of lesion found was determined. In horses that competed in the barrel race, a predominance of lesions in the thoracic caudal, thoracolumbar, and cranial lumbar regions occurred, with intervertebral osteoarthritis and interspinous desmitis being the most common. In cutting horses, most of the lesions were observed in the caudal lumbar region, whereas horses competing in reining showed a preferential location for lesions in the middle lumbar, with a predominance of supraspinous desmitis and myositis. Thermography associated with ultrasonography was shown to be efficient in the diagnosis of the thoracolumbar lesions of these horses.

Clinical applications of non-culture based methods for the diagnosis and management of opportunistic and endemic mycoses

Nonculture based methods for the detection of infections caused by fungal pathogens are becoming more important tools in the management of infected patients. Detection of fungal antigens and DNA appear to be the most promising in this respect for both opportunistic and endemic mycoses. In this article we present an overview of the most recent developments in nonculture based methods and examine their value in clinical practice.

The instantaneous-space-phasor a powerful diagnosis tool

This paper presents the instantaneous-space-phasor (ISP) definition and describes its properties and possible applications for three-phase systems. It is shown that the ISP provides the mathematical base for a new approach to the measurement of active, reactive, and apparent power. Moreover, the ISP helps separate the positive and negative-sequence components and fits perfectly into the Buchholz-Goodhue apparent power definition.