Simultaneous osteosarcoma in two limbs of a Maltese dog

Osteosarcoma (OSA) is rarely diagnosed in small breed dogs and it is usually restricted to a single bone, despite its aggressive biological behavior. This paper aims to report a rare case of OSA in an eight-year-old female spayed Maltese dog with simultaneous presentation in thoracic and pelvic limbs, associated with lung and kidney micrometastasis. Physical exam revealed an increase volume in the distal region of the left femur and in the proximal region of the left humerus with intense pain sensibility at palpation. Pelvic limb´s radiographs revealed an extensive area of bone lysis in the distal femur, lamellated periosteal reaction and formation of the Codman triangle, while in the thoracic limb, it was evident a cortical fracture of the proximal humerus and mixed periosteal reaction, suggestive of neoplastic or infectious disease. Cytological analysis showed the presence of a large number of malignant star-shaped cells, with prominent anisocytosis and anisokaryosis, moderate to prominent cellular and nuclear pleomorphism, rare mitotic figures, besides abundant extracellular matrix compatible with osteoid, suggestive of osteosarcoma. Histopathological findings were compatible with OSA. The dog died of a cardiorespiratory arrest and necropsy revealed a grade II osteoblastic osteosarcoma in the humerus and femur, along with renal metastasis and neoplastic emboli in the kidney and lungs.

Simultaneous osteosarcoma in two limbs of a Maltese dog

Osteosarcoma (OSA) is rarely diagnosed in small breed dogs and it is usually restricted to a single bone, despite its aggressive biological behavior. This paper aims to report a rare case of OSA in an eight-year-old female spayed Maltese dog with simultaneous presentation in thoracic and pelvic limbs, associated with lung and kidney micrometastasis. Physical exam revealed an increase volume in the distal region of the left femur and in the proximal region of the left humerus with intense pain sensibility at palpation. Pelvic limb´s radiographs revealed an extensive area of bone lysis in the distal femur, lamellated periosteal reaction and formation of the Codman triangle, while in the thoracic limb, it was evident a cortical fracture of the proximal humerus and mixed periosteal reaction, suggestive of neoplastic or infectious disease. Cytological analysis showed the presence of a large number of malignant star-shaped cells, with prominent anisocytosis and anisokaryosis, moderate to prominent cellular and nuclear pleomorphism, rare mitotic figures, besides abundant extracellular matrix compatible with osteoid, suggestive of osteosarcoma. Histopathological findings were compatible with OSA. The dog died of a cardiorespiratory arrest and necropsy revealed a grade II osteoblastic osteosarcoma in the humerus and femur, along with renal metastasis and neoplastic emboli in the kidney and lungs.(AU)

Soil pretreatment and fast cell lysis for direct polymerase chain reaction from forest soils for terminal restriction fragment length polymorphism analysis of fungal communities

Humic substances in soil DNA samples can influence the assessment of microbial diversity and community composition. Using multiple steps during or after cell lysis adds expenses, is time-consuming, and causes DNA loss. A pretreatment of soil samples and a single step DNA extraction may improve experimental results. In order to optimize a protocol for obtaining high purity DNA from soil microbiota, five prewashing agents were compared in terms of their efficiency and effectiveness in removing soil contaminants. Residual contaminants were precipitated by adding 0.6 mL of 0.5 M CaCl2. Four cell lysis methods were applied to test their compatibility with the pretreatment (prewashing + Ca2+ flocculation) and to ultimately identify the optimal cell lysis method for analyzing fungal communities in forest soils. The results showed that pretreatment with TNP + Triton X-100 + skim milk (100 mM Tris, 100 mM Na4P2O7, 1% polyvinylpyrrolidone, 100 mM NaCl, 0.05% Triton X-100, 4% skim milk, pH 10.0) removed most soil humic contaminants. When the pretreatment was combined with Ca2+ flocculation, the purity of all soil DNA samples was further improved. DNA samples obtained by the fast glass bead-beating method (MethodFGB) had the highest purity. The resulting DNA was successfully used, without further purification steps, as a template for polymerase chain reaction targeting fungal internal transcribed spacer regions. The results obtained by terminal restriction fragment length polymorphism analysis indicated that the MethodFGB revealed greater fungal diversity and more distinctive community structure compared with the other methods tested. Our study provides a protocol for fungal cell lysis in soil, which is fast, convenient, and effective for analyzing fungal communities in forest soils.(AU)

ISOLAMENTO DE BACTERIÓFAGOS LÍTICOS EM EFLUENTES HOSPITALARES E SUA ATIVIDADE CONTRA PSEUDOMONAS AERUGINOSA E ACINETOBACTER BAUMANNII MULTIRRESISTENTES

Um dos maiores desafios de saúde pública dos dias atuais é o tratamento de infecções severas por bactérias oportunistas multi ou panresistentes, como Pseudomonas aeruginosa e Acinetobacter baumannii. Antibióticos de última linha amplamente utilizados, como os carbapenens, não são mais tão eficazes devido ao desenvolvimento de mecanismos de resistência bacteriana. Outras classes com maior poder de debelar os patógenos, como as glicilciclinas e as polimixinas, também enfrentam o aumento exponencial de resistência, gerando um contingente de pacientes cujas infecções são simplesmente intratáveis. Conhecendo os mecanismos de resistência que essas duas espécies têm desenvolvido ao longo do tempo, é urgente a busca de alternativas ao uso dos antibióticos para o tratamento de infecções por isolados multirresistentes. Dentro dessa perspectiva, o emprego dos bacteriófagos tem se mostrado promissor, pois o desenvolvimento científico-tecnológico atual possibilita o isolamento de fagos líticos vislumbrando seu emprego na terapêutica adequada. O objetivo do presente trabalho foi isolar e caracterizar bacteriófagos líticos a partir de esgoto hospitalar, avaliando sua atividade diante de isolados clínicos de P. aeruginosa e A. baumannii multirresistentes (MDR). O esgoto foi coletado de um hospital localizado no litoral norte do Estado (RS). Os 22 isolados bacterianos oriundos de dois laboratórios tiveram o seu perfil de sensibilidade aos antibióticos determinado segundo Brazilian Committee on Antimicrobial Susceptibility Testing (BrCAST) para checar a condição de MDR. A pesquisa de profago foi realizada para excluir as bactérias com fagos lisogênicos. Os demais isolados de P. aeruginosa e A. baumannii (12) foram expostos ao filtrado do esgoto no ensaio em sobrecamada de ágar, para isolamento de fagos líticos. Laboratorialmente, foram encontrados cinco fagos líticos específicos para P. aeruginosa, oriundos de três diferentes amostras, mas nenhum fago para A. baumannii. Pode-se observar que o esgoto hospitalar é uma fonte interessante para o isolamento de fagos líticos ativos sobre bactérias de interesse clínico, como P. aeruginosa MDR. A caracterização molecular de fagos apropriados para biocontrole de patógenos é essencial para qualquer tratamento à base de bacteriófagos. A perspectiva é que tais fagos possam ter aplicabilidade como agentes para o controle de infecções não-complicadas.

One of the biggest public health challenges of today is the treatment of severe infections by multi or pan-resistant opportunistic bacteria, such as Pseudomonas aeruginosa and Acinetobacter baumannii. Broad spectrum antibiotics, such as carbapenens, are no longer effective due to the development of mechanisms of bacterial resistance. Other classes with greater power to quell pathogens, such as glycylcyclines and polymyxins, also face an exponential increase in resistance, generating a contingent of patients whose infections are simply intractable. Knowing the resistance mechanisms that these two species have developed over time, the search for alternatives to the use of antibiotics for the treatment of infections by multidrug-resistant isolates is urgent. Within this perspective, the use of bacteriophages has been defined as promising, as the current scientific and technological development allows the isolation of lytic phages, envisioning their use in the specialty. The aim of the present study was to isolate and characterize lytic bacteriophages from hospital sewage, evaluating its activity in the presence of multidrug-resistant (MDR) P. aeruginosa and A. baumannii isolates. The sewage collected from a hospital located on the north coast of RS. The 22 bacterial isolates from two laboratories had their antibiotic sensitivity profile determined according to the Brazilian Committee on Antimicrobial Susceptibility Testing (BrCAST) to check the condition of MDR. The prophage search was performed to exclude bacteria with lysogenic phages. The 5 isolates of P. aeruginosa and 7 isolates of A. baumannii were exposed to the sewage filtrate in the agar overlay assay, for isolation of lytic phages. In the laboratory, five specific lytic phages were found for P. aeruginosa, from three different samples, but no phage for A. baumannii. I has been demonstrated that hospital sewage is an interesting source of the isolation of active lytic phages on bacterial of clinical interest, such as P. aeruginosa MDR. Molecular characterization of phages suitable for biocontrol of pathogens is essential for any treatment based on bacteriophages. The perspective is that such phages may have applicability in the treatment of uncomplicated infections.

Production, purification and application of extracellular chitinase from Cellulosimicrobium cellulans 191

This study concerned the production, purification and application of extracellular chitinase from Cellulosimicrobium cellulans strain 191. In shaken flasks the maximum yield of chitinase was 6.9 U/mL after 72 h of cultivation at 25ºC and 200 rpm. In a 5 L fermenter with 1.5 vvm aeration, the highest yield obtained was 4.19 U/mL after 168 h of fermentation at 25ºC and 200 rpm, and using 3 vvm, it was 4.38 U/mL after 144 h of fermentation. The chitinase (61 KDa) was purified about 6.65 times by Sepharose CL 4B 200 gel filtration with a yield of 46.61%. The purified enzyme was able to lyse the cell walls of some fungi and to form protoplasts.

O presente estudo visou a produção, purificação e aplicação da quitinase extracelular da linhagem Cellulosimicrobium cellulans 191. A maior produção de quitinase em frascos agitados foi 6,9 U/mL após 72 h de fermentação a 25ºC e 200 rpm. Em fermentador de 5 L utilizando aeração de 1,5 vvm, a maior atividade da enzima foi 4,19 U/mL após 168 h de fermentação a 25ºC e 200 rpm; e com 3 vvm, foi obtido 4,38 U/mL após 144 h de fermentação. A quitinase (61 KDa) foi purificada cerca de 6,65 vezes em coluna de filtração em gel Sepharose CL 4B 200 com um rendimento de 46,61%. A enzima purificada foi capaz de lisar a parede celular de alguns fungos e formar protoplastos.

Organic carbon determination in histosols and soil horizons with high organic matter content from Brazil

Soil taxonomy systems distinguish mineral soils from organic soils based on the amount of soil organic carbon. Procedures adopted in soil surveys for organic carbon measurement are therefore of major importance to classify the soils, and to correlate their properties with data from other studies. To evaluate different methods for measuring organic carbon and organic matter content in Histosols and soils with histic horizons, from different regions of Brazil, 53 soil samples were comparatively analyzed by the methods of Walkley & Black (modified), Embrapa, Yeomans & Bremner, modified Yeomans & Bremner, muffle furnace, and CHN. The modified Walkley & Black (C-W & B md) and the combustion of organic matter in the muffle furnace (OM-Muffle) were the most suitable for the samples with high organic carbon content. Based on regression analysis data, the OM-muffle may be estimated from C-W & B md by applying a factor that ranges from 2.00 to 2.19 with 95% of probability. The factor 2.10, the average value, is suggested to convert results obtained by these methods.

Sistemas taxonômicos distinguem horizontes e/ou camadas minerais das orgânicas baseando-se na quantidade de carbono orgânico. Assim, o procedimento adotado em pesquisas para a quantificação do conteúdo de carbono orgânico é de grande importância para a classificação das terras e correlacionar as suas propriedades com dados de outros estudos. Com o objetivo de avaliar os diferentes métodos para medir o conteúdo de carbono orgânico e de matéria orgânica em Organossolo e solos com elevados teores de matéria orgânica, de diferentes regiões do Brasil, cinqüenta e três amostras de terra foram comparativamente analisadas pelos métodos de Walkley & Black (modificado), Embrapa, Yeomans & Bremner, Yeomans & Bremner modificado, forno mufla, e CHN. O método Walkley & Black modificado (C-W & B md) e a combustão de matéria orgânica no forno mufla (MO-Mufla) revelaram-se mais satisfatórios para as amostras com elevado conteúdo de carbono orgânico. Com base em dados de análise de regressão, os teores de matéria orgânica determinados pela mufla podem ser calculados a partir dos dados de C-W & B md aplicando-se um fator que varia de 2,00 a 2,19 com 95% de probabilidade. O fator 2,10, valor médio, é sugerido para a conversão dos resultados obtidos por estes métodos.

Organic carbon determination in histosols and soil horizons with high organic matter content from Brazil

Soil taxonomy systems distinguish mineral soils from organic soils based on the amount of soil organic carbon. Procedures adopted in soil surveys for organic carbon measurement are therefore of major importance to classify the soils, and to correlate their properties with data from other studies. To evaluate different methods for measuring organic carbon and organic matter content in Histosols and soils with histic horizons, from different regions of Brazil, 53 soil samples were comparatively analyzed by the methods of Walkley & Black (modified), Embrapa, Yeomans & Bremner, modified Yeomans & Bremner, muffle furnace, and CHN. The modified Walkley & Black (C-W & B md) and the combustion of organic matter in the muffle furnace (OM-Muffle) were the most suitable for the samples with high organic carbon content. Based on regression analysis data, the OM-muffle may be estimated from C-W & B md by applying a factor that ranges from 2.00 to 2.19 with 95% of probability. The factor 2.10, the average value, is suggested to convert results obtained by these methods.

Sistemas taxonômicos distinguem horizontes e/ou camadas minerais das orgânicas baseando-se na quantidade de carbono orgânico. Assim, o procedimento adotado em pesquisas para a quantificação do conteúdo de carbono orgânico é de grande importância para a classificação das terras e correlacionar as suas propriedades com dados de outros estudos. Com o objetivo de avaliar os diferentes métodos para medir o conteúdo de carbono orgânico e de matéria orgânica em Organossolo e solos com elevados teores de matéria orgânica, de diferentes regiões do Brasil, cinqüenta e três amostras de terra foram comparativamente analisadas pelos métodos de Walkley & Black (modificado), Embrapa, Yeomans & Bremner, Yeomans & Bremner modificado, forno mufla, e CHN. O método Walkley & Black modificado (C-W & B md) e a combustão de matéria orgânica no forno mufla (MO-Mufla) revelaram-se mais satisfatórios para as amostras com elevado conteúdo de carbono orgânico. Com base em dados de análise de regressão, os teores de matéria orgânica determinados pela mufla podem ser calculados a partir dos dados de C-W & B md aplicando-se um fator que varia de 2,00 a 2,19 com 95% de probabilidade. O fator 2,10, valor médio, é sugerido para a conversão dos resultados obtidos por estes métodos.

Physico-chemical composition of Apis mellifera L. honey samples of the state of Tocantins, Brazil / Composição físico-química de amostra de méis de Apis mellifera L. do Estado de Tocantins, Brasil

This paper was carried out to determine how can be grouped the Apis mellifera honey samples of the State of Tocantins, Brazil, with base to physico-chemical composition of honey. The samples were collected in different municipalities. Among the 21 honey samples analyzed it was observed that the ones which have Mimosa scabrella, Caesalpinia sp and Arecaceae type ones as dominant or accessory pollen, form different groups or sub-groups based on the physicochemical parameters. The results confirm that the honey bee composition is influenced its origin nectar. The grouping of the honey samples was mostly influenced by viscosity and pH in the X axle, and acidity and reducing sugars, in the Y axle.

A presente pesquisa, desenvolvida com amostras de méis de Apis mellifera coletadas diretamente dos produtores de diversos municípios do Estado de Tocantins, Brasil, teve o objetivo de verificar, com base em características físico-químicas, como se agrupam as amostras de méis. Dentre as 21 amostras de méis analisadas as que continham pólen de Mimosa scabrella, Caesalpinia sp e tipo Arecaceae como dominante ou acessório, formam grupos ou subgrupos distintos quanto aos caracteres físico-químicos, o que confirma que a origem floral interfere decisivamente nas características dos méis. Pela análise dos componentes principais, pode-se verificar que os caracteres que mais influenciaram no agrupamento das amostras de méis foram viscosidade e pH no eixo X e acidez e açúcares redutores no eixo Y.

Sheep erythrocyte sensitisation and restriction to lysis by the complement system from equine and sheep serum / Sensibilização de hemácias de carneiro e restrição à lise pelo sistema complemento do soro eqüino e de carneiro

The effect of the degree of sheep red cell sensitisation by antibody in the resistance to lysis by sheep and horse complement was analysed. The results showed that, as the level of sensitisation increased, so did the homologous complement activity, eventually leading to one hundred percent of lysis; the same was observed for horse serum, generally considered as not efficient at lysing these cells. Therefore, the homologous and heterologous restriction can be overridden by this treatment. This may be of interest for the comprehension of the mechanisms involved in the modulation of the restriction phenomenon in health an in diseases involving complement in domestic animals and in human, as well as in laboratory diagnosis where hemolytic assays are used for measurement of complement activity and evaluation of presence of antibodies in serum samples.

Neste trabalho foi analisado o efeito do grau de sensibilização de hemácias de carneiro por anticorpo, na resistência à lise pelo complemento do soro eqüino e de carneiro. Os resultados mostraram que, à medida que houve um aumento do nível de sensibilização, ocorreu também um aumento da atividade do complemento homólogo, levando a 100% de lise; o mesmo foi observado para o complemento do soro eqüino, geralmente considerado ineficiente na lise destas células. A restrição à lise pode, assim, ser sobrepujada por este tratamento. Estes dados podem ser de interesse para a compreensão dos mecanismos envolvidos na modulação do fenômeno de restrição, na saúde e em doenças envolvendo o sistema complemento em animais domésticos e no homem, bem como em diagnóstico laboratorial, onde são utilizados ensaios hemolíticos para medida de atividade do complemento e pesquisa de anticorpos em amostras de soro.

Physico-chemical composition of Apis mellifera L. honey samples of the state of Tocantins, Brazil / Composição físico-química de amostra de méis de Apis mellifera L. do Estado de Tocantins, Brasil

This paper was carried out to determine how can be grouped the Apis mellifera honey samples of the State of Tocantins, Brazil, with base to physico-chemical composition of honey. The samples were collected in different municipalities. Among the 21 honey samples analyzed it was observed that the ones which have Mimosa scabrella, Caesalpinia sp and Arecaceae type ones as dominant or accessory pollen, form different groups or sub-groups based on the physicochemical parameters. The results confirm that the honey bee composition is influenced its origin nectar. The grouping of the honey samples was mostly influenced by viscosity and pH in the X axle, and acidity and reducing sugars, in the Y axle.

A presente pesquisa, desenvolvida com amostras de méis de Apis mellifera coletadas diretamente dos produtores de diversos municípios do Estado de Tocantins, Brasil, teve o objetivo de verificar, com base em características físico-químicas, como se agrupam as amostras de méis. Dentre as 21 amostras de méis analisadas as que continham pólen de Mimosa scabrella, Caesalpinia sp e tipo Arecaceae como dominante ou acessório, formam grupos ou subgrupos distintos quanto aos caracteres físico-químicos, o que confirma que a origem floral interfere decisivamente nas características dos méis. Pela análise dos componentes principais, pode-se verificar que os caracteres que mais influenciaram no agrupamento das amostras de méis foram viscosidade e pH no eixo X e acidez e açúcares redutores no eixo Y.