A study of "white spotted kidneys" in cattle

A study was performed at an abattoir in Australia, in an attempt to correlate focal chronic interstitial nephritis (FCIN) producing the so-called white spotted kidney, with Leptospira spp. and other pathogens in cattle. Samples of kidneys, urine and blood were collected immediately after slaughter from 46 two-year-old heifers, and 72 cows and bulls with gross lesions consistent with FCIN. The same samples were also collected from nine heifers and 12 cows with no gross kidney lesions. Aqueous humour was also collected from the eye of 17 of the adult animals. The sera were processed by a microscopic agglutination test for leptospira antibodies, while all the other samples were cultured for Leptospira spp. and also processed for routine aerobic and anaerobic culture for other pathogens. Sub-samples from all the kidneys were fixed in 10% buffered formalin and processed histologically. Antibody titers of 1:400 or higher for Lepstospira borgpeterseni serovar hardjo were found in six adult animals with FCIN and in one adult animal with no gross kidney changes, while antibody titers of 1:400 to L borgpeterseni serovar tarassovi were found in only one animal with FCIN. L. borgpeterseni serovar hardjo was isolated from the urine and kidney of one adult animal and from the urine of another adult animal, both with FCIN. No pathogens were isolated from any of the other samples. The histological lesions were consistent in most cases with FCIN. The results suggest that neither Leptospira spp. nor active infection by other bacteria are associated with c the so-called white spotted kidneys. (C) 2002 Elsevier Science B.V. All rights reserved.

Alternative technologies for producing sterile low acid food products

Four emerging high-energy non-thermal technologies may replace or augment heating for producing sterile low-acid food products. High pressure, high-voltage pulsed electric field, high-energy ultrasound and high-intensity pulsed light are all capable of reducing bacterial spore counts under certain conditions. However, only non-continuous high pressure treatments, at temperatures higher than ambient, are currently capable of completely inactivating spores and producing sterile food products. The first three technologies also reduce the resistance of spores to inactivation by heat.

A survey of bacterial diversity in ticks, lice and fleas from Australia

We isolated bacteria from ticks, lice and fleas. Partial small subunit rRNA sequences were obtained for each isolate and the closest matches in the FastA database were determined. These bacteria were mostly Gram-positive (Firmicutes), although representatives from the Proteobacteria (alpha, beta, gamma subdivisions) and CFB group were also isolated. Most of the isolates we found were from genera that were present in most of the ectoparasites studied, but a few genera were restricted to one species of ectoparasite. The most commonly isolated genera were Stenotrophomonas, Staphylococcus, Pseudomonas, Acinetobacter and Bacillus. Species of Bacillus and Proteus, which have biopesticide potential, were found in some of these ectoparasites. Overall, the communities of bacteria were similar to those found in other studies of parasitic arthropods.

Amylases in enzyme detergents and their effects on a simulated long-life dairy dessert

Enzyme detergents used in the food industry contain proteinase as the major enzyme but amylase may be present, either by design or inadvertently. Three commercial enzyme detergents and 3 enzyme preparations used in detergents were assayed for alpha-amylase activity by the Ceralpha method using the Megazyme kits. The amylase activities of the detergents varied from 3.2x 10(-6) to 32x 10(-6) mumoles ml(-1) h(-1) while the enzyme preparations had much higher activities ranging from 0.05 to 8.06 mumoles ml(-1) h(-1). When added aseptically to a simulated dairy dessert (2% starch solution) and stored for 42 days, the enzyme detergents caused an increase in viscosity; enzyme preparations at low concentrations caused an initial increase in viscosity followed by a decrease; and enzyme preparations at high concentrations caused an immediate decrease in viscosity. The increase in viscosity corresponded to formation of a distinct network of starch granules while the decrease in viscosity was characterised by a marked decrease in size of the granules and little or no network of granules. Decreases in viscosity corresponded to increases in reducing sugars but samples which increased in viscosity showed no measurable reducing sugars. The amylase activity in all sources was destroyed by heating at 75degreesC for 15 min at pH 1.8.

Combining nonthermal technologies to control foodborne microorganisms

Novel nonthermal processes, such as high hydrostatic pressure (HHP), pulsed electric fields (PEFs), ionizing radiation and ultrasonication, are able to inactivate microorganisms at ambient or sublethal temperatures. Many of these processes require very high treatment intensities, however, to achieve adequate microbial destruction in low-acid foods. Combining nonthermal processes with conventional preservation methods enhances their antimicrobial effect so that lower process intensities can be used. Combining two or more nonthermal processes can also enhance microbial inactivation and allow the use of lower individual treatment intensities. For conventional preservation treatments, optimal microbial control is achieved through the hurdle concept, with synergistic effects resulting from different components of the microbial cell being targeted simultaneously. The mechanisms of inactivation by nonthermal processes are still unclear; thus, the bases of synergistic combinations remain speculative. This paper reviews literature on the antimicrobial efficiencies of nonthermal processes combined with conventional and novel nonthermal technologies. Where possible, the proposed mechanisms of synergy is mentioned. (C) 2003 Elsevier Science B.V. All rights reserved.

OxyR acts as a repressor of catalase expression in Neisseria gonorrhoeae

It has been reported that Neisseria gonorrhoeae possesses a very high level of catalase activity, but the regulation of catalase expression has not been investigated extensively. In Escherichia coli and Salmonella enterica serovar Typhimurium, it has been demonstrated that OxyR is a positive regulator of hydrogen peroxide-inducible genes, including the gene encoding catalase. The oxyR gene from N. gonorrhoeae was cloned and used to complement an E. coli oxyR mutant, confirming its identity and function. The gene was inactivated by inserting a kanamycin resistance cassette and used to make a knockout allele on the chromosome of N. gonorrhoeae strain 1291. In contrast to E. coli, the N. gonorrhoeae oxyR::kan mutant expressed ninefold-more catalase activity and was more resistant to hydrogen peroxide killing than the wild type. These data are consistent with OxyR in N. gonorrhoeae acting as a repressor of catalase expression.

Sanidade de sementes de arroz, biocontrole, caracterização e transmissão de Curvularia lunata em semente-plântula de arroz

Este trabalho objetivou avaliar a qualidade sanitária de sementes de arroz, quantificar a transmissão de Curvularia lunata associada à semente e à plântula, e avaliar o efeito dos isolados de Bacillus spp., no controle da mancha foliar, do cultivar Bonança. Para avaliação da sanidade de sementes, as amostras foram desinfestadas e plaqueadas, sendo avaliadas após sete dias de incubação. Na confirmação da patogenicidade, as plântulas foram inoculadas por meio de pulverização e a avaliação da doença ocorreu sete dias após. Na caracterização do isolado de C. lunata, foi realizada medição da largura, comprimento e número de septos de 100 conídios. Para a taxa de transmissão de C. lunata, foram preparadas 12 bandejas, com 100 sementes cada, de modo a proceder-se às avaliações aos 7, 14, 21 dias após a semeadura (d.a.s.). De cada uma das 100 plântulas, foram separadas as raízes, após sete dias. A inoculação dos isolados de Bacillus foi realizada na forma de suspensão, pulverizada sobre a parte aérea das plântulas. Após uma semana, foi inoculado a C. lunata. A avaliação foi efetuada aos 15 dias após inoculação dos isolados patogênicos, medindo-se o tamanho da lesão. No teste de sanidade foi detectada maior incidência de C. lunata (70%), o qual se apresentou patogênico à variedade de arroz Bonança, sendo detectado em todos os órgãos da plântula. Dentre os isolados de Bacillus, os que mais se destacaram no controle foram B41 (B. Cereus) e B35 (Bacillus sp.), confirmando seu potencial biocontrolador.

Pesquisa de alguns microrganismos em saladas com maionese adquiridas em restaurantes, lanchonetes e "rotisseries"

Em vinte amostras de saladas com maionese foram efetuadas as contagens de bactérias mesófilas e psicrófilas, Staphylococcus aureus, Bacillus cereus, bolores e leveduras, a determinação do Número Mais Provável (NMP) de bactérias coliforme totais de Escherichia coli e de estreptococos fecais, bem como a pesquisa de salmonelas. A contagem de bactérias mesófilas variou de 2,64 x 10(4) a ³3 x 10(7)/g do produto. Quanto às bactérias psicrófilas, as contagens variaram de < 10 a ³ 3 x 10(7)/g. Para S. aureus, as contagens oscilaram de < 10² a 4 x 10(5)/g do alimento, enquanto que para B. cereus os números mínimo e máximo foram < 10² e ³ 3 x 10(4)/g, respectivamente. Para bolores e leveduras, as contagens variaram de 7,1 x 10² a 3,7 x 10(6)/g. Com relação ao NMP de coliformes totais e estreptococos fecais, os resultados obtidos mostraram-se compreendidos entre < 0,03 e ³ 4,3 x 10(5)/g. Quanto ao NMP de E. coli os números mínimo e máximo obtidos foram respectivamente de < 0,03 e ³ 2,4 x 10(4)/g de salada com maionese. Tais constatações indicam a ocorrência de contaminação inclusive por microrganismos de origem fecal. Todas as amostras revelaram-se negativas para bactérias do gênero Salmonella.

Esterilização por óxido de etileno: II. Influência de corpos de prova no desempenho de monitores biológicos e sua avaliação

Pela importância da garantia de esterilidade em produto médico-hospitalares e pela predominância da forma tubular dentre estes produtos, conduziu-se o estudo de monitores biológicos em que os corpos de prova apresentavam diferentes comprimentos e diâmetros. O sensor biológico foi constituído de esporos de Bacillus subtilis var. niger adquirido do mercado nacional sob tiras de papel. Após esterilização por processo industrial, seguiu-se a recuperação dos esporos sobreviventes através de inoculação dos suportes em caldo tioglicolato, caldo caseína-soja e este adicionado de azul de bromotimol. A capacidade promotora de crescimento destes 3 meios de cultura não apresentou diferenças. A eficácia esterilizante foi dependente do tamanho de corpos de prova. A determinação periódica do teor residual de gás nos corpos de prova, bem como a monitoração ambiental industrial indicaram a importância da legislação em vigor, devendo ser obedecida a fim de garantir inocuidade aos pacientes e operadores.

Cellular uptake of BCG-loaded chitosan microparticles and in vivo evaluation of immune response following intranasal immunisation

Attenuated Mycobacterium bovis bacillus Calmette-Guérin (BCG) is the only currently available vaccine against tuberculosis. It is highly effective in pre-exposure immunisation against TB in children when administered by subcutaneous route to newborns. However, it does not provide permanent protection in adults. In this work, polymeric chitosan-alginate microparticles have been evaluated as potential nasal delivery systems and mucosal adjuvants for live attenuated BCG. Chitosan (CS) has been employed as adjuvant and mucosal permeation-enhancer, and, together with alginate (ALG), as additive to enhance BCG-loaded microparticles (MPs) cellular uptake in a human monocyte cell line, by particle surface modification. The most suitable particles were used for vaccine formulation and evaluation of immune response following intranasal immunisation of BALB/c mice.

A new natural spiro heterocyclic compound and the cytotoxic activity of the secondary metabolites from Juniperus brevifolia leaves

Copyright © 2010 Elsevier B.V. All rights reserved.

Protein-ligand docking study: diterpenes from Juniperus brevifolia as anticancer and antimicrobial agentes

REDCAT: Natural Products and related Redox Catalysts: Basic Research and Applications in Medicine and Agriculture, Aveiro, 25-27 Novembro de 2012.

Potencial da diversidade microbiana dos Açores na saúde : screening, purificação e caracterização de uma enzima trombolítica

Dissertação de Mestrado, Mestrado em Ciências Biomédicas, 8 de Maio de 2015, Universidade dos Açores.

Macrophages as biomarkers in BCG treatment response in bladder cancer

Bladder cancer is a common urologic cancer and the majority has origin in the urothelium. Patients with intermediate and high risk of recurrence/progression bladder cancer are treated with intravesical instillation with Bacillus Calmette-Guérin, however, approximately 30% of patients do not respond to treatment. At the moment, there are no accepted biomarkers do predict treatment outcome and an early identification of patients better served by alternative therapeutics. The treatment initiates a cascade of cytokines responsible by recruiting macrophages to the tumor site that have been shown to influence treatment outcome. Effective BCG therapy needs precise activation of the Th1 immune pathway associated with M1 polarized macrophages. However, tumor-associated macrophages (TAMs) often assume an immunoregulatory M2 phenotype, either immunosuppressive or angiogenic, that interfere in different ways with the BCG induced antitumor immune response. The M2 macrophage is influenced by different microenvironments in the stroma and the tumor. In particular, the degree of hypoxia in the tumors is responsible by the recruitment and differentiation of macrophages into the M2 angiogenic phenotype, suggested to be associated with the response to treatment. Nevertheless, neither the macrophage phenotypes present nor the influence of localization and hypoxia have been addressed in previous studies. Therefore, this work devoted to study the influence of TAMs, in particular of the M2 phenotype taking into account their localization (stroma or tumor) and the degree of hypoxia in the tumor (low or high) in BCG treatment outcome. The study included 99 bladder cancer patients treated with BCG. Tumors resected prior to treatment were evaluated using immunohistochemistry for CD68 and CD163 antigens, which identify a lineage macrophage marker and a M2-polarized specific cell surface receptor, respectively. Tumor hypoxia was evaluated based on HIF-1α expression. As a main finding it was observed that a high predominance of CD163+ macrophage counts in the stroma of tumors under low hypoxia was associated with BCG immunotherapy failure, possibly due to its immunosuppressive phenotype. This study further reinforces the importance the tumor microenvironment in the modulation of BCG responses.

Response of high-risk of recurrence/progression bladder tumours expressing sialyl-Tn and sialyl-6-T to BCG immunotherapy

High risk of recurrence/progression bladder tumours is treated with Bacillus Calmette-Guérin (BCG) immunotherapy after complete resection of the tumour. Approximately 75% of these tumours express the uncommon carbohydrate antigen sialyl-Tn (Tn), a surrogate biomarker of tumour aggressiveness. Such changes in the glycosylation of cell-surface proteins influence tumour microenvironment and immune responses that may modulate treatment outcome and the course of disease. The aim of this work is to determine the efficiency of BCG immunotherapy against tumours expressing sTn and sTn-related antigen sialyl-6-T (s6T). METHODS: In a retrospective design, 94 tumours from patients treated with BCG were screened for sTn and s6T expression. In vitro studies were conducted to determine the interaction of BCG with high-grade bladder cancer cell line overexpressing sTn. RESULTS: From the 94 cases evaluated, 36 had recurrence after BCG treatment (38.3%). Treatment outcome was influenced by age over 65 years (HR=2.668; (1.344-5.254); P=0.005), maintenance schedule (HR=0.480; (0.246-0.936); P=0.031) and multifocality (HR=2.065; (1.033-4.126); P=0.040). sTn or s6T expression was associated with BCG response (P=0.024; P<0.0001) and with increased recurrence-free survival (P=0.001). Multivariate analyses showed that sTn and/or s6T were independent predictive markers of recurrence after BCG immunotherapy (HR=0.296; (0.148-0.594); P=0.001). In vitro studies demonstrated higher adhesion and internalisation of the bacillus to cells expressing sTn, promoting cell death. CONCLUSION: s6T is described for the first time in bladder tumours. Our data strongly suggest that BCG immunotherapy is efficient against sTn- and s6T-positive tumours. Furthermore, sTn and s6T expression are independent predictive markers of BCG treatment response and may be useful in the identification of patients who could benefit more from this immunotherapy.