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In addition to Staphylococcus aureus nowadays other coagulase-positive staphylococci (CoPS) and coagulase-negative staphylococci (CoNS), earlier considered of minor importance, are now accepted as relevant pathogens for humans and animals. The involvement of these microorganisms in bovine mastitis etiology and the possibility their transmission through milk to humans justify the requirement of developing reliable methods for identification of the most frequent species among them. The purpose of this study was to compare the phenotypic techniques with the genotypic method carried out by sequencing of the rpoB gene in identification of several species of the genus Staphylococcus isolated from bovine mastitis. A total of 300 staphylococci isolates of bovine mastitis cases from several Brazilian dairy herds were studied by phenotypic and genotypic techniques, respectively: 150 CoPS and 150 CoNS strains. A total of 18 CoNS different species and 4 CoPS species were identified. Among the CoNS the following species were recognized: 48 (32%) Staphylococcus warneri, 22(15%) S. epidermidis, 20(13%) S. hyicus, 10(7%) S. xylosus, 7(5%) S. haemolyticus, 6(4%) S. simulans, 6(4%) S. schleiferi subsp schleiferi, 6(4%) S. hominis, 5(3%) S. pasteuri, 4(2.7%) S. cohnii, 3(2%) S. saprophyticus subsp. saprophyticus 3(2%) S. chromogenes 3(2%) S. sciuri, 2(1%) S. saccharolyticus, 2(1%) S. lugdunensi, 1(0,7%) S. auricularis, 1(70%) S. saprophyticus subsp. bovis, 1(0.7%) S. capitis. And among the 150 CoPS were identified respectively: 105 (70%) S. aureus, 21(14%), S. hyicus, 19(13%) S. intermedius e 5(3%) S. schleiferi subsp coagulans. Considering the 150 CoNS isolates, the identifications performed by phenotypic and genotypic tests presented 96.7% of concordance, kappa coefficient of agreement = 0.933, SE (standard error) of kappa=0.021 (95% confidence interval: 0.893 to 0.974), Pearson’s correlation coefficient (r) = 0.9977, (confidence interval 95%: 0.9938 a 0.9992) and in relation to 150 CPS isolates it was detected an agreement of 98.7%, kappa = 0.960, SE of kappa = 0.016, (95% confidence interval: 0.929 to 0.992) Pearson’s correlation coefficient (r) = 0.9994 (95% confidence interval: 0.9681 to 1.0000). The verified agreement strength between the identification methods can be considered as excellent. These results assure that according to laboratory resources any of them will be suitable to perform the staphylococci identification. RESUMO: Além de Staphylococcus aureus atualmente outros estafilococos coagulase positiva (SCP) e estafilococos coagulase-negativos (SCN), anteriormente considerados de menor relevância, são reconhecidos como importantes patógenos para humanos e animais. O envolvimento desses micro-organismos na etiologia da mastite bovina e a possibilidade da sua transmissão através do leite aos humanos justifica a utilização de métodos confiáveis para a identificação das espécies mais frequentes. O objetivo deste estudo foi comparar as técnicas fenotípicas com o método genotípico realizada por sequenciamento do gene rpoB na identificação de espécies do gênero Staphylococcus spp. isolados de mastite bovina. Um total de 300 estafilococos isolados de casos de mastite bovina em diferentes rebanhos leiteiros brasileiros foram estudados por técnicas fenotípicas e genotípicas, respectivamente: 150 linhagens de SCP e 150 linhagens de SCN. Foram identificados um total de 18 espécies de SCN e 4 espécies SCP. Entre os SCN as seguintes espécies identificadas: 48 (32%) Staphylococcus warneri, 22 (15%) S. epidermidis, 20 (13%) S. hyicus, 10 (7%) S. xylosus, 7 (5%) S. haemolyticus, 6 (4%) S. simulans, 6 (4%) S. schleiferi subsp schleiferi, 6 (4%) S. hominis, 5 (3%) S. pasteuri, 4 (2,7%) S. cohnii, 3 (2%) S. saprophyticus subsp. saprophyticus, 3 (2%) S. chromogenes, 3 (2%) S. sciuri, 2 (1%) S. saccharolyticus, 2 (1%) S. lugdunensi, 1 (0,7%) S. auricularis, 1 (70 %) S. saprophyticus subsp. bovis, 1 (0,7%) S. capitis. E entre as 150 SCP foram identificados, 105 (70%) S. aureus, 21 (14%), S. hyicus, 19 (13%) S. intermedius e 5 (3%) S. schleiferi subsp coagulans. Considerando-se os 150 SCN isolados, as identificações realizadas por testes fenotípicos e genotípicos apresentaram 96,7% de concordância, coeficiente de concordância kappa = 0,933, SE (erro padrão) de kappa = 0,021 (95% intervalo de confiança: 0,893-0,974), coeficiente de correlação de Pearson (r) = 0,9977, (intervalo de confiança de 95%: 0,9938 a 0,9992) e em relação a 150 SCP isolados foi observado uma concordância de 98,7%, kappa = 0,960, sE de kappa = 0,016, (95% de intervalo de confiança: 0,929 a 0,992) coeficiente de correlação de Pearson (r) = 0,9994 (95% intervalo de confiança: 0,9681-1,0000). A correlação entre os métodos de identificação pode ser considerada como excelente. Esses resultados demonstraram que de acordo com os recursos disponíveis no laboratório, poderia ser utilizada qualquer uma das metodologias.

A dual-chambered microbial fuel cell with aqueous cathode was operated with domestic wastewater to investigate the electrogenic ability of anaerobic bacteria from a municipal wastewater treatment plant. Curves of cell potential vs. current density, power density vs. current density and current at a fixed load of 100 Ω, were obtained daily to monitor the electrochemical evolution of the system as a function of substrate use in several batch cycles. A maximum power density of 1.11 μWcm-2 was obtained after 65 days of continuous operation and a coulombic efficiency of 7% and a chemical oxygen demand removal of 76% were found in the last batch cycle. Anaerobic culture of the bacteria from the anode biofilm resulted in the isolation of two Gram-positive and two Gram-negative bacteria with divergent sugar fermentation capabilities, while analysis of 16S rRNA gene fragments showed three clones from the phyla Firmicutes, δ-Proteobacteria and α- Proteobacteria. Scanning electron imaging analysis revealed an increase in cell diversity and proliferation of methanogenic archaea when changing from synthetic to real wastewater. These results reveal the influence of substrate concentration and presence of methanogenic microorganisms on the production of power in microbial fuel cells, suggesting that future developments could be a contribution for the use of this technology to decontaminate domestic wastewater in small communities. Resumen Una celda de combustible microbiana de doble cámara con cátodo inmerso en el agua fue operada con aguas residuales domésticas para investigar la capacidad electrogénica de las bacterias anaeróbicas de una planta de tratamiento de aguas residuales municipales. Se obtuvieron diariamente curvas de potencial de celda vs. densidad de corriente, densidad de potencia vs. densidad de corriente y corriente a una resistencia fija de 100 Ω para monitorear la evolución electroquímica del sistema en función del uso del sustrato en varios ciclos de lotes. Se obtuvo una densidad de potencia máxima de 1.11 µWcm-2 después de 65 días de operación continua, una eficiencia coulombic del 7% y una disminución de la Demanda de Oxígeno Químico del 76% en el último ciclo del lote. El cultivo anaeróbico de las bacterias de la biopelícula anódica dio como resultado el aislamiento de dos bacterias Gram-positivas y dos Gram-negativas con capacidades de fermentación de azúcar divergentes, mientras que el análisis de los fragmentos del gen 16S rRNA mostró tres clones de los filos Firmicutes, Prote-Proteobacteria y α -Proteobacterias. El análisis de las imágenes de microscopia electronica de barrido reveló un aumento en la diversidad celular y la proliferación de arqueas metanogénicas al cambiar de aguas residuales sintéticas a reales. Estos resultados revelan la influencia de la concentración de sustrato y la presencia de microorganismos metanogénicos en la producción de energía en la celda de combustible microbiana, lo que sugiere que los desarrollos futuros podrían ser una contribución para el uso de esta tecnología para descontaminar las aguas residuales domésticas en las comunidades rurales.

La Organización Mundial de la Salud (OMS) define salmonelosis como una de las enfermedades transmitida por alimentos (ETA) de mayor casuística, ampliamente extendida en todo el mundo. La enfermedad es producida por Salmonella spp. y causa una de las zoonosis más frecuentes y de mayor impacto económico. El hombre adquiere la infección después de la ingestión de alimen-tos contaminados, aunque también puede transmitirse de persona a persona o por vía fecal-oral. Actualmente, las técnicas micro-biológicas de aislamiento convencional para detección de Salmonella spp. son establecidas por el Código Alimentario Argentino para verificar la aptitud de un producto para consumo, pero éstas requieren de 4 a 5 días para la obtención de un resultado, tiem-po que juega en contra para el productor y la conservación de dichos alimentos. Por este motivo en este trabajo se analizan los métodos de diagnóstico tradicional según Normas ISO 6579:2002 con algunas modificaciones, los métodos de inmunoensayo comerciales y la Reacción en Cadena de la Polimerasa técnica (PCR) de detección del gen invA implicado en el proceso de inva-sión de cepas patógenas. Se analizaron 60 muestras procedentes de canales porcinas destinadas a comercialización. Se detectó un 10% de Salmonella spp. Se pudo determinar que el diagnóstico molecular por PCR posee alta sensibilidad, pero no es alenta-dor el resultado que reflejan los test comerciales inmunocromatográficos ya que queda en evidencia la necesidad de alta carga microbiana para un diagnóstico certero. The World Health Organization (WHO) defines salmonellosis as one of the most important foodborne diseases, widely spread worldwide. The disease is produced by Salmonella spp. and causes one of the most frequent zoonoses and of greater economic impact. The infection is acquired after ingestion of contaminated food, although it can also be transmitted from person to person or by fecal-oral route. Currently, conventional isolation microbiological techniques for detection of Salmonella spp. are established by the Argentine Food Code to verify the suitability of a product for consumption. But microbiological techniques require 4 to 5 days to obtain a result, time that plays against the producer and the conservation of such foods. For this reason in this work we analyze the traditional diagnostic methods according to ISO standards 6579: 2002 with some modifications, the commercial immunoassay methods and the Polymerase Chain Reaction technique (PCR) detection of the invA gene involved in the process of invasion of pathogenic strains. Sixty samples from pigs destined for commercialization were analyzed. 10% of Salmonella spp. was detected. It was possible to determine that the molecular diagnosis by PCR has high sensitivity, but it is not encouraging the result that reflect the commercial immunochromatographic tests since it is evident the need of high microbial load for a correct diagnosis. Fil: Ruiz, María Julia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. Departamento de Sanidad Animal y Medicina Preventiva. Laboratorio de Inmunoquímica y Biotecnología; Argentina Fil: Ramallo, Grisel. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. Departamento de Sanidad Animal y Medicina Preventiva. Laboratorio de Inmunoquímica y Biotecnología; Argentina Fil: Colello, Rocío. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. Departamento de Sanidad Animal y Medicina Preventiva. Laboratorio de Inmunoquímica y Biotecnología; Argentina Fil: Villalobo, Maria Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. Departamento de Sanidad Animal y Medicina Preventiva. Laboratorio de Inmunoquímica y Biotecnología; Argentina Fil: Monteavaro, Cristina Esther. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. Departamento de Sanidad Animal y Medicina Preventiva. Laboratorio de Inmunoquímica y Biotecnología; Argentina Fil: Etcheverría, Analía Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. Departamento de Sanidad Animal y Medicina Preventiva. Laboratorio de Inmunoquímica y Biotecnología; Argentina Fil: Padola, Nora Lía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. Departamento de Sanidad Animal y Medicina Preventiva. Laboratorio de Inmunoquímica y Biotecnología; Argentina

Simbiozni odnos kvržičnih bakterija i mahunarki ima važnu ulogu u poljoprivrednoj proizvodnji. Veliku važnost ima odabir sojeva koji su otporni na stresne uvjete u tlu. Danas se sve više susrećemo s ekstremnim vremenskim uvjetima te je od velike važnosti selekcija visoko učinkovitih sojeva kvržičnih bakterija, kako bi predsjetvena bakterizacija sjemena mahunarki bila uspješna. Glavni cilj rada je utvrditi fenotipske karakteristike sojeva kvržičnih bakterija izoliranih iz kvržica soje u uvjetima koji su izvan optimalnih vrijednosti za njihov rast. Sposobnost rasta sojeva istražen je na različitim pH vrijednostima, temperaturama, koncentracijama soli, izvorima ugljikohidrata i testirana je osjetljivost na određene antibiotike. Provedena su oksidaza, ureaza, i katalaza testovi, ispitana je mobilnost fosfata, sposobnost korištenja citrata te hidroliza škroba. Istražene su morfološke karakteristike izolata, te otpornost na fitopatogene gljive iz roda Phomopsis. Uzorci tla za izolaciju sojeva su prikupljeni s područja Koprivničko – križevačke, Bjelovarsko – bilogorske i Vukovarsko – srijemske županije. Istraživanjem je obuhvaćeno 18 izolata simbioznih fiksatora dušika soje, te pet tipskih i jedan referenti soj. Utvrđeno je da su svi ispitani izolati gram negativne, štapićaste bakterije koje ne sadrže kapsule. Rezultati su pokazali da većina ispitanih izolata tolerira rast u lužnatim i kiselim sredinama te mogu rasti na temperaturi od 37 °C. Svi izolati pokazali su dobar rast na različitim izvorima ugljikohidrata (galaktoza, laktoza, dekstroza, saharoza, maltoza, rafinoza i fruktoza), dok su referentni sojevi većinom pokazali znatno slabiji rast. Izolati sa područja Koprivničko – križevačke županije jedini su pokazali mogućnost rasta na podlogama koje sadrže značajno povećani sadržaj soli (1 %, 2 % i 3 % NaCl-a). Oksidaza testom dokazano je da 40 % izolata posjeduje enzim citokromoksidazu. Sve bakterije koje posjeduju enzim citokromoksidazu koriste kisik kao terminalni akceptor elektrona u disanju. Sposobnost hidrolize škroba pokazalo je 60 % ispitanih izolata. Jedan od osnovnih problema koji donosi povećanje površina i intenzivna proizvodnje soje svakako je problem bolesti i to prije svega bolesti uzrokovanih fitopatogenim gljivama na koje otpada oko 80 % svih bolesti soje. U istraživanju antagonističke aktivnosti izolata prema pojedinim gljivama 6 od 18 izolata pokazalo otpornost na fitopatogene gljive iz roda Phomopsis. Dobiveni rezultati ukazuju na značaj selekcije autohtonih sojeva. The symbiosis of nodule bacteria and legumes plays a very important role in agricultural production. It is essential that the highly effective strains that are resistant to various unfavorable conditions in the soil are chosen. Today we are increasingly faced with extreme weather, so the selection of highly efficient strains of rhizobia is of great importance to the success of legumes seed inoculation. The aim of this study is to determine the phenotypic characteristics of nodule bacteria isolated from soybean nodules under conditions that are beyond the optimum value for their growth. The growth of the strains was studied at different pH values, temperatures, concentrations of salts, carbohydrate sources and resistance to antibiotics. The oxide, the urease, and catalase of test and of phosphate mobility were made. Furthermore, the ability to use citrate and starch hydrolysis was investigated. The resistance to phytopathogenic fungus isolates from genus Phomopsis and the morphologic characteristics were performed as well. The soil samples were collected from the counties Koprivničko – križevačka, Bjelovarsko – bilogorska and Vukovarsko – srijemska. Eighteen isolates of symbiotic soybean fixators, five types and one referent strain were included in the study. It was found that all the tested isolates were gram negative, stinging bacteria that did not contain capsules. Most of the tested isolates tolerated alkaline and acidic environment and they can be grown at the temperature of 37 °C. Furthermore, all the isolates showed good growth on different sources of carbohydrates (galactose, lactose, dextrose, sucrose, maltose, raffinose and fructose), and on the other hand, the reference strains showed a much weaker growth. The isolates from the area of Koprivnica - Križevci County were the only ones that showed the possibility of growth on substrates containing significantly increased salt content (1 %, 2 % and 3 % NaCl). Forty percent of the tested isolates possess the cytochrome oxidase enzyme by examining the test oxidase. All bacteria that possess the cytochrome oxide enzyme use oxygen as the terminal electron acceptor in breathing. The ability of starch hydrolysis showed 60 % of the tested isolates. One of the basic problems which occurs more intensive soybean production is definitely a disease problem. First of all, diseases were caused by phytopathogen funguses, causing about 80 % of all soybean disease. In the study of the antagonistic activity of isolates, according to individual funguses, 6 of all 18 isolates showed the resistance to the phytopathogen funguses Phomopsis. The obtained results indicate the importance of selection of autochthonous strains.

76 p, The purpose of the present study was to contribute to the recognition of the diversity of microorganisms present in the residue lignocellulosic from the process of benefit of the sisal leaf Furcraea andina, the samples were taken at the experimental farm Jose Exelino Moreno, located in the municipality of Mogotes, Santander. The isolation of the culturable microorganisms was given through the use of specific culture media, after performing serial dilutions up to 10-6, later the microbial groups associated whit this residue were established by means of morphological keys and biochemical tests and finally the abundance of microorganisms were determined for each of the samples collected. We obtained a total of 55 isolates correpondian to 36 bacteria, 6 fungi and 13 yeast. In the macroscopic and microscopic characterization, 20 bacteria were selected 70% of the bacteria were Gram negatives and 30% Gram positive. Subsequently whit biochemical tests it could be deduced that there may be the presence of the family Enterobacteriaceae and of the genera Pseudomonas sp. and Bacillus sp. In the same way for the fungi, 7 isolates were selected, of which according to the technique of micro-culture and the taxonomic keys, one finds in greater proportion the genus Penicillium sp.. Finally, we selected 8 isolates of yeast, which according to the results obtained in the API 20 AUX, is the presence of two microorganisms of industrial interest Candida famata y Rhodotorula glutinis, the other yeasts according to the literature are reported as pathogenic for humans. Finally, the isoletes characterized were retained for the purpose of maintaining this biological material in a viable and stable state by method tube whit angle agar in mineral oil., El propósito del presente estudio fue contribuir al reconocimiento de la diversidad de microorganismos presentes en el residuo lignocelulósico proveniente del proceso de beneficio de la hoja de fique Furcraea andina; las muestras fueron tomadas en la finca experimental José Exelino Forero, ubicada en el municipio de Mogotes, Santander. El aislamiento de los microorganismos cultivables se dio miente el uso de medios de cultivo específicos después de realizar diluciones seriadas hasta 10-6, posteriormente se establecieron los grupos microbianos asociados a este residuo mediante claves morfológicas realizando tinción de Gram y pruebas bioquímicas como oxidación y fermentación de azucares y pruebas enzimáticas, por último se determinó la abundancia de microorganismos por cada una de las muestras recolectadas mediante el uso de recuento de unidades formadoras de colonia por mililitro. Se logró obtener un total de 55 aislados correspondientes a 36 bacterias, 6 hongos y 13 levaduras. En la caracterización macroscópica y microscópica, se seleccionaron 20 bacterias 70% de las bacterias fueron Gram negativa y 30% Gram positiva, con las pruebas bioquímicas se pudo deducir que presuntivamente puede existir la presencia de bacterias de la familia Enterobacteriaceae y de los géneros Pseudomonas sp. y Bacillus sp. De igual manera para los hongos se seleccionaron 6 aislados, de las se encuentra en mayor proporción el género Penicillium sp.. Por último, se seleccionaron 8 cepas de levaduras, las cuales según resultados obtenidos en el API 20 AUX, se encuentra la presencia de dos microorganismos de interés industrial, Candida famata y Rhodotorula glutinis, las demás levaduras según literatura son reportadas como patógenas para los humanos. Finalmente, los aislados caracterizadas se conservaron con el propósito de mantener este material biológico en un estado viable y estable, mediante la conservación por método tubo con agar inclinado en aceite mineral., Pregrado, Microbiólogo Industrial, 1. INTRODUCCIÓN ............................................................................................... 11 2. PLANTEAMIENTO DEL PROBLEMA ............................................................. 13 3. PREGUNTA DE INVESTIGACIÓN ................................................................. 14 4. JUSTIFICACIÓN ................................................................................................ 15 5. MARCO TEÓRICO Y BIBLIOGRÁFICO ........................................................ 16 5.1 CARACTERÍSTICAS DEL FIQUE ........................................................... 16 5.2 PROCESO DE BENEFICIO DEL FIQUE .................................................... 17 5.3 PRODUCCIÓN A NIVEL NACIONAL. .......................................................... 20 5.4 COMPOSICIÓN DEL FIQUE Y SU IMPACTO AMBIENTAL. .................. 21 5.5 DESARROLLO TECNOLÓGICO E INVESTIGACIONES ......................... 23 5.6 AGENDA DE INVESTIGACIÓN Y DESARROLLO TECNOLÓGICO DE LA CADENA PRODUCTIVA DE FIQUE 2009-2018 COLOMBIA .................. 24 5.7 COMUNIDADES MICROBINAS Y LA IMPORTANCIA DE ESTUDIARLAS ....................................................................................................... 25 5.8 DIVERSIDAD MICROBIANA EN SUELOS Y PLANTAS .......................... 27 La biodiversidad es la riqueza de organismos vivos de un ecosistema, así como los complejos ecológicos de los que forman parte; comprende la variación dentro de cada especie, entre las especies y los ecosistemas (de la Cruz-Leyva, Zamudio Maya, Corona Cruz, Gonzales de la Cruz , & Rojas Herrera, 2014). El conocimiento de la biodiversidad requiere considerar los diferentes niveles jerárquicos de organización de la vida (genes, especies, poblaciones, comunidades y ecosistemas), junto con sus atributos de composición, estructura y funcionalidad............................................................. 27 6. OBJETIVOS ........................................................................................................ 30 6.1 OBJETIVO GENERAL .................................................................................... 30 6.2 OBJETIVOS ESPECÍFICOS ......................................................................... 30 7. METODOLOGÍA ................................................................................................. 31 7.1 AISLAMIENTO DE MICROORGANISMOS PRESENTES EN EL RESIDUO LIGNOCELULÓSICO Y CARACTERIZACIÓN MORFOLÓGICA 32 SIEMBRA Y RECUPERACIÓN DE MICROORGANISMOS ........................... 32 7.2 CARACTERIZACIÓN BIOQUÍMICA PARA BACTERIAS Y LEVADURAS. ......................................................................................................... 35 7.3 MÉTODO DE CONSERVACIÓN .................................................................. 35 8. RESULTADOS Y DISCUSIÓN ........................................................................ 37 8.1 AISLAMIENTO DE MIRCROOGANISMOS PRESENTES EN EL RESIDUO LIGNOCELULOSICO ......................................................................... 37 8.2 CARACTERIZACIÓN MACROSCÓPICA Y MICROSCÓPICA ................ 38 8.2.1 CARACTERIZACIÓN MACROSCÓPICA Y MICROSCÓPICA PARA BACTERIAS.................... 38 8.2.2 CARACTERÍSTICAS MACROSCÓPICAS Y MICROSCÓPICAS DE LOS HONGOS................... 42 8.3 BIOQUÍMICA .................................................................................................... 49 8.3.1 CARACTERIZACIÓN BIOQUÍMICA PARA LAS BACTERIAS ......... 49 8.4 CARACTERIZACIÓN PARA LEVADURAS ................................................ 54 8.5 CONSERVACIÓN DE AISLADOS ................................................................ 55 9. CONCLUSIONES ............................................................................................... 56 10. RECOMENDACIONES ................................................................................. 56 11. BIBLIOGRAFÍA ............................................................................................... 58 12. ANEXOS .......................................................................................................... 64, Ej. 1

BACKGROUND : Tuberculosis is considered to be a major cause of death among the infectious diseases prevalent in India. Differentiation of M. tuberculosis from Nontuberculous Mycobacteria (NTM) is very important in the management of the disease. Recently, Standard Diagnostics (SD Yongin Korea) developed a simple and rapid assay using a mouse monoclonal anti - MPT64 antibody to discriminate between M. tuberculosis and NTM by Immu no - chromatography. These tests were found to have good sensitivity and specificity. METHOD : 200 sputum samples were processed and inoculated on conventional Lowenstein Jensen culture medium. The Mycobacterial isolates were subjected to biochemical tests (Ni acin test and Nitrate reduction test) and Immunochromatographic assay. The isolates were also tested by Polymerase chain reaction. H37 RV strain was used as a positive control. RESULT : The culture isolates showed positive in 47 (23.5%) cases. 44 isolates w ere positive and 3 were negative by both biochemical tests and immunochromatographic assay. Similar results were obtained by PCR. The sensitivity and specificity of Immunochromatography test when compared to biochemical tests and PCR was 100%. Positive pre dictive value and Negative predictive values were 100%. CONCLUSION : Immunochromatographic assay showed comparable results with biochemical tests and this assay can be used as an alternative to biochemical tests for the confirmation of Mycobacterium tubercu losis complex, which avoids use of hazardous chemicals (Cyanogen bromide).

Different types of anticoagulants are used in the routine of a veterinary clinic, which are chosen based on the exam to be performed. Agitated, aggressive, of hard contention and small size animals can be a limiting factor when it is wished to separate samples into tubes with different anticoagulants. Therefore, this study had as the objective evaluate the effects of anticoagulants: EDTA, sodium citrate and sodium fluorite associated with EDTA, in the results of routine plasma biochemistry performed in dogs and compare it with serum. Blood samples of 10 clinically healthy dogs were collected and placed in different tubes with each anticoagulant and one without to obtain serum. Albumin, alanine aminotransferase (ALT), calcium, total cholesterol, creatine, phosphorus, glucose, total protein (PT) and urea concentrations were measured by spectophotometric. It was found that plasma obtained with EDTA anticoagulant resulted in dosage alterations for glucose, cholesterol and calcium. Samples obtained with sodium citrate presented results lower than serum in all analyzed parameters, except for ALT. The anticoagulant sodium 34 Colloquium Agrariae, v. 11, n.1, Jan-Jun. 2015, p.33-41. DOI: 10.5747/ca.2015.v11.n1.a116 fluorite with EDTA only didn’t result in statistical differences when compared to serum in the parameters of urea and glucose. Thus, serum is the most trustworthy sample to determine the biochemical profile of dogs, however some biochemical’s can be performed with plasma with EDTA such as creatine, ALT, urea, albumin, PT and phosphorus.

O sorgo é um cereal de grande importância comercial. Todavia, essa gramínea não se adapta bem a solos de baixa fertilidade tornado a cultura dependente do uso de fertilizantes industriais. Na busca para solucionar esse problema, a os benefícios que podem ser proporcionados interação entre bactérias associadas a plantas têm sido objeto de estudo de vários trabalhos de pesquisa. No entanto, atualmente existem poucos estudos sobre bactérias isoladas do sorgo. O objetivo deste trabalho foi realizar a caracterização polifásica (morfológica, bioquímica e genética) de bactérias endofíticas isoladas de raízes de sorgo. Foram selecionadas um total de 24 isolados usando o meio semisseletivo de NFb-lactato, LGI-P e YMA. A caracterização foi baseada nos aspectos de morfológicos; bioquímicos com ênfase no potencial para produção de enzimas e genético pela análise da região intergênica 16S-23S rRNA. Todos os isolados foram classificados como gram-negativo e apresentaram grande resistência a antibióticos e ampla flexibilidade metabólica quanto ao uso de diferentes fontes de carbono. A maioria dos isolados (95,8%) mostrou atividade enzimática para de catalase, urease, redutase de nitrato e nitrogenase. Entre as enzimas hidrolíticas, as bactérias isoladas de sorgo apresentaram maior potencial para produção de lipases (70,8%) e celulases (50%). Mais de 90% dos isolados foram capazes de solubilizar o fosfato. A análise de similaridade permitiu identificar uma grande diversidade morfofisiológica e genética entre as bactérias isoladas das raízes de sorgo. Foi possível ainda constatar que vários isolados apresentaram mecanismo relacionado à promoção do crescimento da planta e podem ser considerados promissores como inoculantes ou como novas fontes de enzimas para uso biotecnológico. Sorghum is a cereal of great commercial importance. However, this grass does not adapt well to soils of low fertility making the crop dependent on the use of industrial fertilizers. In the quest to address this problem, the benefits that can be provided for interaction between bacteria associated with plants have been the subject of study of various research papers. However, there are currently few studies on bacteria isolated from sorghum. The objective of this work was to characterize polyphase (morphological, biochemical and genetic) of endophytic bacteria isolated from sorghum roots. A total of 24 isolates were selected using the NFb-lactate, LGI-P and YMA semi-insulin medium. The characterization was based on morphological aspects; biochemicals, directed to enzymatic potential and genetic by PCR of the 16S-23S rRNA intergenic region. All isolates were classified as gram-negative and showed high resistance to antibiotics and wide metabolic flexibility regarding the use of different carbon sources. Most of the isolates (95.8%) showed enzymatic activity for catalase, urease, nitrate reductase and nitrogenase. Among the hydrolytic enzymes, the bacteria isolated from sorghum presented higher potential to produce lipases (70.8%) and cellulases (50%). More than 90% of the isolates solubilize phosphate. The analysis of similarity allowed identify morpho-physiological and genetic diversity between the bacteria isolated from the roots of sorghum. It was also possible to verify isolates with mechanisms related to the promotion of plant growth, which can be considered as promising as inoculants or as new sources of enzymes for biotechnological use. Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES

RESUMO: Este trabalho avaliou a influência da infecção bacteriana da glândula mamária (GM) sobre a transferência de imunidade passiva (TIP) em bezerros recém-nascidos. Vacas holandesas (n=13) foram observadas no momento da parição e ordenhadas para a obtenção de forma asséptica das amostras de colostro (n=52) para os testes microbiológicos. Os recém-nascidos receberam 6 litros de colostro de uma ordenha nas primeiras 12 horas de vida, proveniente de suas respectivas mães. Amostras de sangue foram colhidas antes (D0) e após (D2) o manejo do colostro. A TIP foi avaliada por meio de testes bioquímicos, eletroforese e leucograma. Os bezerros foram distribuídos conforme a ausência (IB-) ou presença (IB+) de infecção mamária em pelo menos uma GM de suas respectivas mães. Todas as amostras de colostro (n=52) foram negativas ao cultivo fúngico. Das 13 fêmeas, 8 (61%) apresentaram crescimento bacteriano em ≥1 quartos mamários. Considerando-se os quartos mamários, foi obtido isolamento bacteriano em 21,15% (11/52), observando-se predomínio de espécies bacterianas do grupo Staphylococcus coagulase negativa. Não foram encontradas diferenças entre os parâmetros de acordo com os grupos experimentais. Em relação aos momentos, foi possível verificar aumento nos valores de proteína total, globulinas, atividade sérica da gama glutamiltransferase e frações eletroforéticas beta e gamaglobulina após a ingestão do colostro materno. A mastite subclínica não influencia a transferência de imunidade passiva em bezerros recém-nascidos da raça holandesa, avaliados por teste bioquímicos, eletroforese e leucograma. ABSTRACT: This study evaluated the influence of intramammary infection under the passive immunity transfer in newborn calves. Holstein cows (n=13) were monitored at delivery, and milked to obtain sterile colostrum samples (n=52) for microbiological examination. The newborn calves received 6 liters of colostrum from their dams up to 12 hours after birth. Blood samples were harvested before (D0) and after (D2) colostrum feeding. The passive immune transfer was evaluated by biochemical tests, electrophoresis and leukogram. Calves were distributed according to absence (IB-) or presence (IB+) of mammary gland infection in one or more quarters. All colostrum samples (n=52) were negative for fungal culture. Eight dams (8; 61%) had bacterial growth in one or more mammary quarter. In relation to mammary gland unit, bacterial growth was observed in 21.15% (11/52) of the colostrum samples, with predominance of coagulase negative staphylococcal infections. Statistical differences were not found among variables according to the groups. According to the experimental moments, the evaluated parameters (total protein, albumin, globulins, GGT and beta and alpha electrophoretic) increased after colostrum intake. Subclinical mastitis did not influence the passive immune transfer in Holstein calves evaluated by biochemical tests, electrophoresis or leukogram.

Aeromonas hydrophila is an opportunistic microorganism. It is a secondary biological agent that contributes to the occurrence of fish diseases and its deterioration. This research was undertaken to determine the prevalence of A. hydrophila in some freshwater fishes collected from three different fish markets of Dhaka City and to test their antibiotic susceptibility. Total bacterial count and total aeromonas on different aeromonas selective media were enumerated using serial dilution technique. Bacterial isolates were characterized to identify A. hydrophila using biochemical tests and with comparison to reference strain (ATCC 7966). The lowest Aeromonas count was detected to be 2.83±0.40×10 2 cfu/g in Anabas testudineus and the highest was 1.03±0.153×10 3 cfu/g in Oreochromis mossambicus . On market basis highest aeromonas count was found in Anando Bazar (8.10±1.09×10 2 cfu/g) and lowest in Hatirpool Bazar (5.63±0.90×10 2 cfu/g) with no significant difference. Maximum susceptibility to amikacin and gentamicin was observed whereas all of the isolates were found resistant to a commonly used antibiotic amoxycillin. The obtained results point that antimicrobial susceptibility was more or less similar regardless of the origin of the samples collected. All the fishes investigated in this study contained A. hydrophila in their different organs.

La caña de azúcar es un cultivo importante en Colombia, intensivamente cultivado en el valle del río Cauca en cerca de 237.070 hectáreas. Para alcanzar altas productividades, se aplican altas y algunos casos excesivas dosis de fertilizantes nitrogenados, incrementando los costos de producción y agudizando los efectos ambientales. En la búsqueda de estrategias para reducir las aplicaciones de N dentro de un manejo integrado del cultivo, el uso de bacterias fijadoras de N está siendo explorado. Así entonces, el objetivo de este estudio fue aislar y caracterizar bacterias nativas fijadoras de N y promotoras de crecimiento pertenecientes a los géneros Azospirillum, Azotobacter y Gluconoacetobater en cuatro variedades comerciales sembradas en el valle del río Cauca (CC85-92, CC84-75, CC93-4418 y CC01-1940), en 108 lotes o suertes seleccionados para este estudio. Las muestras fueron compuestas a partir de 20 plantas (tallos, hojas, raíces y suelo rizosférico) por sitio, seleccionadas aleatoriamente, para un total de 2.160 plantas, las cuales se transportaron y procesaron en el Laboratorio de Fitopatología de CENICAÑA. De los extractos vegetales se inocularon los medios de cultivo selectivos LGI-P, Ashby y NFB para realizar el aislamiento y caracterización para cada género. Se obtuvieron 111 aislamientos caracterizados morfológica y bioquímicamente agrupados con base en pruebas por triplicado para α and β-proteobacterias, incluyendo aerobiosis/microaerofílicos, motilidad, tinción de Gram, metabolismo oxidativofermentativo y metabolismo de diferentes carbohidratos y sales orgánicas. De los 111 aislamientos, se identificaron molecularmente 15 aislamientos bacterianos del género Azospirillum spp., 32 aislamientos del género Gluconacetobacter spp. y 20 aislamientos del género Azotobacter spp. mediante secuenciación del gen 16S del ADN ribosomal. La cuantificación de ácido 3-indol acético y reducción de acetileno mostró diferencias significativas en producción de fitohormonas y en la eficiencia de la fijación de nitrógeno, obteniendo cinco cepas bacterianas (Números 2, 14, 16, 21 y 24) que presentaron una alta capacidad fijadora de nitrógeno y producción de compuestos indólicos con respecto a las cepas de referencia de Azotobacter chroococcum NCBIM 8002, Azospirillum brasilense NCBIM 11860, Gluconacetobacter diazotrophicus NCBIM 12985. La secuenciación y alineamiento del gen nifH para muestras contrastantes de algunos de estos aislamientos no presentó variantes genéticas entre los aislamientos con mayor fijación biológica de nitrógeno, indicando una alta conservación de este gen. Se identificaron 14 aislamientos de los tres géneros como candidatos promisorios para trabajos y estudios futuros. //Abstract: Sugarcane is an important crop in Colombia, intensively planted in nearly 237,070 hectares along the Cauca’s river valley. To reach profitable yields, high and in some cases excessive nitrogen (N) fertilizers are applied, increasing production costs and risks for environmental and water contamination. Looking for strategies to reduce N applications within a sustainable and integrated crop management, the use of N-fixing bacteria is being explored. The objective of this study is to isolate and characterize native cultures of three recognized N-fixing bacteria and plant growth promoter genera such as Azospirillum, Azotobacter, and Gluconoacetobater, associated to CC 85-92, CC 84-75, CC 93-4418, and CC 01-1940 varieties, which are widely planted in this region. To cover most environments along the valley, 108 fields were selected to take composed samples from roots, stems, leaves, and rhizospheric soil from 20 plants randomly selected per field, for a total of 2,160 plants in this study. Samples were processed at CENICAÑA’s Plant Pathology Lab, plating on selective culture media LGI-P, Ashby, and NFB, among others for each bacteria genus. 111 isolates were morphologically and biochemically characterized based on triplicated tests for α and β-proteobacteria, including aerobic/microaerophilic, motility- helical-shaped rods and cocci, Gram stain, and oxidative-fermentative metabolism of different carbohydrates and organic salts. Out of 111 isolates, were identified 15 isolates for Azospirillum spp., 32 for Gluconacetobacter spp. y 20 isolates for Azotobacter spp. By sequencing of 16S ribosomal DNA. Quantitation of Indole-3 Acetic Acid as growth promoting substance and acetylene reduction as nitrogen fixation indicator, showed significant differences between genera and isolates, obtaining five isolates (Numbers. 2, 14, 16, 21 and 24) with high N- fixing and indolic compounds producing capability and efficiency in comparison to reference published isolates type cultures of Azotobacter chroococcum NCBIM 8002, Azospirillum brasilense NCBIM 11860, Gluconacetobacter diazotrophicus NCBIM 12985, among other bacteria. Sequencing and alignment of nifH gene region for contrasting samples of several of these isolates did not show genetic differences between isolates with higher nitrogen fixation efficiency, indicated high conservation of this gene. Fourteen isolates from the three genera were identified as promising candidates for further studies. Maestría

The paper deals with the issue of the influence of soldiers' nutrition in foreign missions on the contents of total cholesterol (TCH), triglyceride (TAG), glucose, and alanine aminotranferase (ALT). Combined rations consisting of the preparation of hot meals on a military base and combat rations of foodstuffs that provide soldier's nutrition, in case of impossibility to deliver the hot meals, are an optimal solution. Adverse effect on consumption had psychosocial factors, i.e. eating in a hurry and stress, about the culture of eating it cannot be spoken at all. Air pollutants of chemical contamination and air contamination of the diet had a significant negative impact. The biochemical tests showed that the prolonged duration of the mission led to deterioration of health, especially in dislipidemic disorders. The increase in levels of TCH is particularly evident with the troops in the KFOR mission. The obtained values are characterized as an increased risk of acute cholesterolemia and are also enhanced by the low average age of evaluated group. The growing trend of increase in cholesterolemia is presented as a general phenomenon by foreign and domestic authors.

Gelam honey exerts anti-inflammatory and antioxidant activities and is thought to have potent effects in reducing infections and healing wounds. The aim of this study was to investigate the effects of intravenously-injected Gelam honey in protecting organs from lethal doses of lipopolysaccharide (LPS). Six groups of rabbits (N = 6) were used in this study. Two groups acted as controls and received only saline and no LPS injections. For the test groups, 1 mL honey (500 mg/kg in saline) was intravenously injected into two groups (treated), while saline (1 mL) was injected into the other two groups (untreated); after 1 h, all four test groups were intravenously-injected with LPS (0.5 mg/kg). Eight hours after the LPS injection, blood and organs were collected from three groups (one from each treatment stream) and blood parameters were measured and biochemical tests, histopathology, and myeloperoxidase assessment were performed. For survival rate tests, rabbits from the remaining three groups were monitored over a 2-week period. Treatment with honey showed protective effects on organs through the improvement of organ blood parameters, reduced infiltration of neutrophils, and decreased myeloperoxidase activity. Honey-treated rabbits also showed reduced mortality after LPS injection compared with untreated rabbits. Honey may have a therapeutic effect in protecting organs during inflammatory diseases.

Background and Objectives: Isolation of Xanthomonas campestris from soil has a wide range of applications from monitoring of phytopathogenic populations in soil to screening of improved xanthan-producing strains. Identification of Xanthomonas campestris and its pathovars requires pathogenicity tests in addition to phenotypic and molecular characterization. Materials and Methods: Thirty phenotypic tests were carried out on 57 yellow-pigmented bacterial isolates obtained from soil of cabbage farms after screening on Selective Xanthomonas (SX) agar and transferring on Yeast Malt agar. Absorption spectra of pigments and capability of biopolymer production were determined for the isolates. Some characteristics of the biopolymer produced and presence of a X. campestris-specific gene marker were investigated for nine putative X. campestris isolates. Results: The present study introduces a set of simple phenotypic tests including urease, acid production from sucrose, mucoid growth on 5% sucrose, starch hydrolysis, growth in 4% NaCl, motility and utilization of asparagine as sole carbon and nitrogen source for quick and inexpensive tentative identification of Xanthomonas campestris. Validation of these tests was confirmed in 100% of the cases by characterization of bacterial exopolysaccharide as xanthan and production of genus-specific xanthomonadin pigment. Moreover, tracking of hrc gene among putative X. campestris isolates gave positive results in 80% of cases. Conclusion: The Minimal simple phenotypic tests facilitate the screening and differentiation of putative X. campestris isolates from other false bacterial strains isolated from soil on semiselective SX agar.

Evaluation of a simplified key for the identification of coagulase-positive Staphylococcus isolated from bovine mastitis. Three hundred fourty four strains of coagulase-positive Staphylococcus (CPS), isolated from mastitis cases, underwent phenotypic and genotypic tests to evaluate the efficiency of a simplified key, based on phenotypic tests for the discrimination of these microorganisms. The tests consisted of amplification of the femA gene and hemolysis in blood agar, production of acetoin and fermentation of maltose, mannitol and trehalose. Strains that showed negative results in the amplification test of the femA gene or that were not identified as Staphylococcus aureus (S. aureus) by phenotypic tests were tested with the APISTAPH kit (Biomériux-France), for precise identification of species. Phenotypic tests revealed 338 strains (98.25%) as S. aureus, three strains (0.86%) as Staphylococcus hyicus, and three microorganisms (0.86%) as Staphylococcus intermedius. PCR demonstrated that 338 (98.25%) strains belonged to the S. aureus species, confirming the results for 336 strains from 338 identified, through a simplified phenotypic key. A high rate of correlation (98.83%) was verifeid between the results of genotypic and phenotypic tests for the identification of S. aureus, demonstrating the applicability of the proposed key, for the discrimination of this microorganism in CPS isolated from bovine mastitis.Evaluation of a simplified key for the identification of coagulase-positive Staphylococcus isolated from bovine mastitis. Three hundred fourty four strains of coagulase-positive Staphylococcus (CPS), isolated from mastitis cases, underwent phenotypic and genotypic tests to evaluate the efficiency of a simplified key, based on phenotypic tests for the discrimination of these microorganisms. The tests consisted of amplification of the femA gene and hemolysis in blood agar, production of acetoin and fermentation of maltose, mannitol and trehalose. Strains that showed negative results in the amplification test of the femA gene or that were not identified as Staphylococcus aureus (S. aureus) by phenotypic tests were tested with the APISTAPH kit (Biomériux-France), for precise identification of species. Phenotypic tests revealed 338 strains (98.25%) as S. aureus, three strains (0.86%) as Staphylococcus hyicus, and three microorganisms (0.86%) as Staphylococcus intermedius. PCR demonstrated that 338 (98.25%) strains belonged to the S. aureus species, confirming the results for 336 strains from 338 identified, through a simplified phenotypic key. A high rate of correlation (98.83%) was verifeid between the results of genotypic and phenotypic tests for the identification of S. aureus, demonstrating the applicability of the proposed key, for the discrimination of this microorganism in CPS isolated from bovine mastitis.

Background: Diabetes mellitus is a metabolic disease leading to abnormal fat, carbohydrate and protein metabolism. Reduced salivary flow rate caused by hyperglycemia is characteristic mainly for periods of poor metabolic control of diabetes, thereby facilitating the growth of aciduric bacteria and caries-lesion development. The objective of our study was to evaluate the effects of diabetes mellitus on dental caries micro-organisms responsible for caries. Materials and Methods: This study was carried out on 60 subjects consisting of 2 groups. The Group A (study group) consisted of 30 subjects with diabetes mellitus and dental caries, and the Group B (control group) consisted of 30 subjects with dental caries but no systemic disease. DFS/dfs index in all subjects was evaluated and compared. Unstimulated salivary flow was collected and levels of Streptococcus mutans were analyzed. Results: It was found that the fasting blood sugar in Group A subjects because of which there was increased streptococcus mutans count and hence high caries index as compared to that of Group B. Conclusion: From our study, we could conclude that with increased age, blood sugar levels, DMFT values, dental caries increases in diabetics than in normal (control) subjects and therefore relationship does exist between diabetis mellitus, oral microbiota and dental caries.

Clostridium perfringens is an important pathogen that provokes numerous different diseases. This bacterium is classified into five different types, each of which capable of causing a different disease. There are various methods for the bacterial identification, many are labor-intensive, time-consuming, expensive and also present low sensitivity and specificity. The aim of this research was to identify the different types of C. perfringens using PCR molecular method. In this study, 130 sheep-dung samples were randomly collected from areas around the city of Kerman, southeastern Iran. After processing and culturing of samples, the produced colonies were morphologically studied, gram stain test was also carried out and the genera of these bacteria were identified through biochemical tests. DNA extracted from isolated bacteria for genotyping was tested by multiplex PCR with specific primers. Based on length of synthesized fragments by PCR, toxin types and bacterial strains were detected. C. perfringens isolated types were divided as follows: 17.39% type A, 21.74% type B, 34.78% type C and 26.09% type D. It should be emphasized that, up to the present moment, C. perfringens type A has not been reported in Iran.

Neurological dysfunction is an important manifestation of inherited metabolic disorders. Although these are more common in childhood, adult onset forms with a different clinical presentation are often encountered. Recent advances in the diagnosis and treatment of these conditions have substantially improved the outcome in many of these conditions. This makes it essential that the practicing physician be familiar with the clinical presentation and diagnosis of these disorders. For the evaluation of a patient with a possible inborn error of metabolism, simple screening tests may aid in the diagnosis and provide direction for more comprehensive laboratory analysis. In this review, we present a practical approach to diagnosis of neurometabolic disorders. Establishing a specific diagnosis in these disorders will enable the clinician in offering a definitive long-term treatment, prognosis and genetic counselling.

Laboratory aids are extensively used in the diagnosis of diseases, in preventive medicine, and as management tools. Reference values of clinically healthy people serve as a guide to the clinician in evaluating biochemical parameters. Determination of 21 biochemical parameters of healthy persons using standard methods of analysis. Cluster analysis and principal components analysis were applied on the above 21 biochemical parameters data. The application of a typical classification approach as cluster analysis proved that four major groups of similarity between all 21 clinical parameters are formed, which correspond to the authors assumption of the existence of several summarizing pattern of clinical parameters such as “enzyme,” “major component excretion”, “general health state,” and “blood specific” pattern. These patterns appear also in the subsets obtained by separation of the general dataset into “male”, “female”, “young”, and “adult” healthy groups. The results obtained from principal components analysis have additionally proved the validity of a similar assumption. The intelligent data analysis on the clinical parameter dataset has shown that when a complex system is considered as a multivariate one, the information about the system substantially increases. All these results support an idea that probably a general health indicator could be constructed taking into account the existing classification groups in the list of clinical parameters.

The accuracy of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for identifying Streptococcus suis isolates obtained from pigs, wild animals, and humans was evaluated using a PCR-based identification assay as the gold standard. In addition, MALDI-TOF MS was compared with the commercial multi-tests Rapid ID 32 STREP system. From the 129 S. suis isolates included in the study and identified by the molecular method, only 31 isolates (24.03%) had score values ≥2.300 and 79 isolates (61.24%) gave score values between 2.299 and 2.000. After updating the currently available S. suis MALDI Biotyper database with the spectra of three additional clinical isolates of serotypes 2, 7, and 9, most isolates had statistically significant higher score values (mean score: 2.65) than those obtained using the original database (mean score: 2.182). Considering the results of the present study, we suggest using a less restrictive threshold score of ≥2.000 for reliable species identification of S. suis. According to this cut-off value, a total of 125 S. suis isolates (96.9%) were correctly identified using the updated database. These data indicate an excellent performance of MALDI-TOF MS for the identification of S. suis.

Benefits of physical activity are undeniable. The aim of the present research was to determine the effects of physical activity and age on cholesterol and glucose levels in the blood, as well as changes in the functional parameters of the cardiovascular system, during stepwise increases in physical load for men employed in the same place, but with different levels of physical activity. The subjects were 95 military officers who were divided into groups according to the level of physical activity of their occupation, with veloergometry used as physical load. Cholesterol and glucose levels in the blood were taken as biochemical indices. The results showed that occupational physical activity had a positive effect on biochemical and cardiovascular functional parameters before, during, and after the physical load. Only the cardiovascular rate (systolic blood pressure) in older subjects was significantly higher than that of the younger persons; for all other parameters, age had no effect at all.

Carbapenem resistance are intrinsic in a few clinical bacteria, such as Stenotrophomonas maltophilia, Aeromonas sp., and Acinetobacter baumannii due to the production of carbapenemases. In addition, most non-fermenters are naturally resistant only to ertapenem and Serratia sp. and Proteeae have intrinsically low-level resistance to imipenem. Indeed, not all carbapenem-resistant isolates produce a carbapenemase (resistance can be mediated by other mechanisms, such as the combination of ESBL plus AmpC, plus impermeability) and not all carbapenemase producers are resistant to carbapenems (enzyme expression at low-level). Concerns about carbapenemase-mediated carbapenem resistance, imply that clinically-significant Gram-negative bacteria should be screened routinely for carbapenem susceptibility. However, detection of carbapenemase production can became difficult, because some carbapenemase-producing isolates demonstrate susceptible carbapenem MICs, even closely to intermediate breakpoint [1]. The ECOFF values, defined by EUCAST, can be a good strategy to detect those isolates [2]. Meropenem offers the best compromise between sensitivity and specificity in terms of detecting carbapenemase-producers, while ertapenem has high sensitivity but low specificity, and therefore not recommended for routine use [3]. Hence, following detection of reduced susceptibility to carbapenems in routine susceptibility tests, specific phenotypic methods for carbapenemases should be applied. The combination disk test, using boronic acid, dipicolinic acid and cloxacillin as β-lactamase inhibitors, is a well-validated choice. In addition, other methods may also be considered for detecting carbapenemase producers, such as Modified Hodge Test (MHT), MALDI-ToF or the new biochemical tests Carba NP and Blue-Carba. Further characterization of the carbapenemase-encoding genes, should include molecular biology methods. info:eu-repo/semantics/publishedVersion

The Liver Unit at the "Dr. José E. González" University Hospital and School of Medicine of the Autonomous University of Nuevo León in Monterrey was founded in 1983. Over the years, it has become a referral center for the northeast of Mexico. The frequency of diagnosis has changed: in 1983, the most common liver disease seen was alcoholic liver disease, today it is chronic hepatitis C. Amebic liver abscess, which used to be common, was hardly seen in 2001. Non-alcoholic fatty liver disease was unidentified 18 years ago, whereas in 2001 it was seen in 10% of patients. The development of five laboratories within the unit has allowed us to implement basic and clinical research trials, and to offer a high quality diagnostic service. The experimental liver transplant program started in 1987 and a clinical program in humans in 1991: four patients received an orthotopic liver transplantation in its first phase. In the second phase, 20 patients received allografts from September 1999 to March 2002. Technical complications have been encountered in only one patient, with a biliary leak, and there have been three perioperative deaths. Infections occurred in eight patients; all resolved. Acute postoperative rejection occurred in two patients, and in the first seven months in another five; all of them resolved. The two-year survival rate is 80%. This unit offers a highly specialized diagnosis, standardized specialized laboratory services and a transplant program that guarantees a higher quality of medical attention to patients with liver diseases.

Objectives: Purulent pericarditis is a collection of purulent effusion in the pericardial space. It has become a rare entity with the increased availability and use of antibiotics. In contrast to pleural empyema, there are few data regarding the biochemical parameters of purulent pericardial effusion to aid diagnosis. Therefore, in this study, we have evaluated the diagnostic utility of biochemical tests in patients with purulent pericarditis. Methods: Between September 2004 and September 2012, we treated fifteen children with purulent pericarditis and tamponade. There were 8 boys and 7 girls, ranging in age from 8 months to 14 years, with a mean age of 5.3 ± 3.2 years. Echocardiographic diagnosis of cardiac tamponade was made in all patients. All patients underwent immediate surgical drainage due to cardiac tamponade. The diagnosis of purulent pericarditis was supported by biochemical tests. Anterior mini-thoracotomy or subxiphoid approach was performed for surgical drainage. Results: The most common clinical findings were tamponade, hepatomegaly, tachycardia, fever refractory antibiotic therapy, dyspnea, tachypnea, cough, and increased jugular venous pressure. Central venous pressure decreased and arterial tension increased immediately after the evacuation of purulent effusion during operation in all patients. The pericardial effusion had high lactic dehydrogenase, and low glucose concentration, confirming purulent pericarditis. Also, pH (mean± SD) was 7.01 ± 0.06. The culture of pericardial effusions and blood samples were negative. Conclusion: Biochemical tests are useful guideline when assessing the pericardial effusions. However, these tests should be interpreted with the clinical and operative findings.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior The use of iron perchlorate as a marker of injury in potato tubers may help preventloss of lots during storage and transport. The objectives of this study were to determine the concentration of iron perchlorate solution and the immersion time for the marking of pectolytic bacteria in potato tubers; the evaluation of potato batches for the presence of these bacteria; the biochemical, physiological and molecular characterization of Pectobacterium spp. isolates; and the indication of pectobacteria by iron perchlorate in inoculated and commercial batches of potato tubers. The experiments were done at the Universidade Federal de Uberlândia, from 2009 to 2011. Marking of pectolytic bacteria in potato tubers was done using iron perchlorate concentrations of 0, 8, 21 and 44%, in which a sample of 10 tubers were immersed for 10, 20 or 30 minutes, and evaluated after 10, 20 and 30 minutes at room temperature, evaluating the appearance dark spots on the tubers. The isolates were characterized culturally, biochemically and physiologically by the tests of: Gram, glucose fermentation, catalase, oxidase, growth at 37ºC, reduction of substances from sucrose, phosphatase activity, indol production and acid production from sorbitol, melibiose, citrate or lactose, hypersensibility reaction in tobacco and pathogenicity in potato baits. Molecular characterization was done using the primers: EXPCCR/EXPCCF; ECA1f/ECA2r, 1491f, L1RA/L1RG Br1f. The indication of pectobacteria by the product studied in inoculated seed potato tubers was done with the cultivars Ágata and Atlantic. The tubers were perforated, or not, with a needle and immersed in distilled water or in bacterial suspension (isolate UFU A14 of Pectobacterium spp.) for 5 minutes and incubated at different temperatures (22, 28 or 35°C) and times (0, 24, 48, 72 or 96 hours), and subsequently immersed in the de perchlorate solution for the evaluation of lesion diameter. The experimental design was randomized blocks with four repetitions, as split plots in space 5 x 4. The indication of pectobacteria by the product in commercial potato batches was done with cultivars Ágata and Asterix. The samples with six tubers were evaluated 12 and 24 hours after harvest and incubated in five temperatures (8, 18, 28, 38 or 48°C), and immersed in 8% iron perchlorate solution for 10 minutes, removed and left at room temperature for 20 minutes before evaluation with a rating scale, varying from 0 to 4. Immersion for 10 minutes in iron perchlorate at 21% and evaluation after 20 minutes at room temperature was adequate to indicate the presence of pectobacteria. The isolates could not be characterized biochemistry and molecular. The inoculation test was able to confirm the presence of pectobacteria in the tubers by perchlorate. Storage and transport of commercial potatoes should be done at lower temperatures; the cultivar Ágata was more susceptible than Asterix to the pectobacteria. O uso de percloreto de ferro como sinalizador de injúrias em tubérculos de batata poderá ajudar evitando perdas de lotes durante o armazenamento e transporte. Os objetivos do trabalho foram determinar a concentração da solução de percloreto de ferro e tempo de imersão para a sinalização de bactérias pectolíticas em tubérculos de batata; a qualificação de lotes de batata quanto à presença destas bactérias; a caracterização bioquímica, fisiológica e molecular de isolados de Pectobacterium spp. e indicação de pectobactérias por percloreto de ferro em tubérculos de batata inoculados e lotes comerciais. Os experimentos foram conduzidos na Universidade Federal de Uberlândia, entre os anos de 2009 e 2011. Para a sinalização de bactérias pectolíticas em tubérculos de batata, foram utilizadas as concentrações 0,0; 8,4; 21,0 e 44,0% de percloreto de ferro, em que amostras de 10 tubérculos ficaram imersas por 10, 20 e 30 minutos. Em seguida, avaliou-se, aos 10, 20 e 30 minutos e em temperatura ambiente, o surgimento de manchas escuras nos tubérculos. Os isolados foram caracterizados cultural, bioquímica e fisiologicamente pelos testes: Gram, fermentação de glicose, catalase, oxidase, crescimento à 37ºC, redução de substâncias a partir de sacarose, atividade da fosfatase, produção de indol e produção de ácido a partir de sorbitol, melibiose, citrato e lactose, reação de hipersensibilidade em fumo e patogenicidade em isca de batata. Para a caracterização molecular, foram utilizados os primers: EXPCCR/EXPCCF; ECA1f/ECA2r, 1491f, L1RA/L1RG Br1f. Para a indicação de pectobactérias pelo produto estudado em tubérculos de batata-semente inoculados, utilizou-se as cvs. Ágata e Atlantic. Os tubérculos foram perfurados e não perfurados com estilete e imersos em água destilada ou na suspensão bacteriana (isolado UFU A14 de Pectobacterium spp.) por 5 minutos e incubados em diferentes temperaturas (22°C, 28°C e 35°C) e tempos (0, 24, 48, 72 e 96 horas); em seguida, imersos em solução de percloreto de ferro com avaliação do diâmetro da lesão formada. O delineamento foi em blocos casualizados com quatro repetições, em esquema de parcelas subdivididas no espaço 5 x 4. Para a indicação de pectobactérias pelo produto em lotes comerciais de batata, foram utilizadas as cultivares Ágata e Asterix. As amostras com seis tubérculos foram avaliadas as 12 e 24 horas após a coleta e incubadas em cinco temperaturas (8°C, 18°C, 28°C, 38°C e 48°C), em seguida, foram imersas por 10 minutos na solução de percloreto de ferro a 8%, retiradas e deixadas em temperatura ambiente por 20 minutos para avaliação através de uma escala de notas variando de 0 a 4. Os tubérculos de batatas foram imersos por 10 minutos na solução de percloreto de ferro 21% e avaliados após 20 minutos no ambiente o que foi suficiente para indicar a presença de fitobactérias. Os isolados não puderam ser caracterizados bioquimica e molecularmente. O teste de inoculação conseguiu comprovar a presença de pectobactérias nos tubérculos pelo percloreto de ferro. Para os lotes comerciais, as menores temperaturas são mais indicadas para o armazenamento e transporte, e a cv. Ágata foi mais suscetível que Asterix. Doutor em Agronomia

In my Bachelor´s thesis I dealt with the issue of investigation of biochemical markers, which are used for diagnostics of metabolic syndrome along with measurement of waist circumference and level of blood pressure. In the theoretical part I defined the term metabolic syndrome. Chapters about glucose and blood lipids tests as well as methods used for their determination are described in detail. Great attention was also paid to preanalytical factors, which significantly share of the accuracy of laboratory test. Practical part of the Bachelor´s thesis is focused on acquisition of practical skills in work with biological material in the laboratory of clinical biochemistry and acquisition of principals of laboratory methods. Under professional supervision in the biochemical laboratory Laboma s.r.o. in České Budějovice, I tested 100 samples of venous blood, where I determined concentration of fasting glucose, triglycerides and HDL cholesterol. The measurement was carried out on the biochemical analyzer Cobas Integra 800 by the method of absorption spectrophotometry. Measured laboratory values were statistically processed and they were interpreted after consultation with the supervisor of the Bachelor´s thesis. The laboratory tests showed that the criteria of metabolic syndrome applied for 21% of women and 15% of men.

Bu çalışma 2-5 Ekim 2011 tarihinde Amsterdam’da düzenlenen VI. Uluslararası Moleküler Böcek Bilimi Sempozyumu’nda poster olarak sunulmuş ve özet olarak basılmıştır. Bu çalışmada Stethorus gilvifrons (Muls.) (Col.:Coccinellidae)un bir hassas (SSS) ve bir de daha az duyarlı (RSS) popülasyonunda bazı insektisitlere ve akarisitlere duyarlılıklarının saptanması yanında, bu popülasyonların duyarlılıklarıyla ilişkili karboksilesteraz (CarE), sitokrom P450 monooksigenaz (P450) and glutatyon S-transferaz (GST) aktiviteleri değerlendirilmiştir. Ayrıca, herbir ilacın bir sublethal ayırıcı dozu (~LD20) ile sinerjist maddeler olan S,S,S-tributilfosforotrithioate (DEF), piperonil butoksit (PBO) ve dietil maleyat (DEM) ile karıştırılarak, muhtemel fizyolojik tolerans mekanizmaları test edilmiştir. Bu popülasyonlarda paraoxon ve pirimicarb inhibitörleri kullanılarak, asetilkolinesteraz (AchE)ın duyarsızlığı saptanmıştır. SSS popülasyonunun LC50 değerleri ile karşılaştırıldığında, RSS popülasyonu abamectin, bifenazate, chlorpyrifos, cyhexatin, hexythiazox ve λ-cyhalothrine sırasıyla 2.1, 11.2, 6.7, 1.7, 1.6 ve 2.3-kat daha az duyarlı bulunmuştur. Sonuçta, S. gilvifronsun dirençli bahçe popülasyonu 4 ilaca orta düzeyde bir duyarlılık kaybı göstermiştir. Substrat ortamı olarak α-naftil asetat ve β- naftil asetat kullanıldığında, CarE aktivitesi RSSde SSSye oranla sırasıyla 1.5 ve 4.3 kat fazla olmuştur. Ayrıca, 1-chloro-2,4- dinitrobenzene ve 2,4- dinitro-1-iodobenzene substarları kullanıldığında GSTlerin aktivitesi RSS popülasyonunda önemli olarak sırasıyla 4.3 ve 1.4 kat daha fazla bulunmuştur. İnhibasyon kinetiklerine göre, 1mM primicarb kullanıldığında RSSnin AchEi SSSye göre 2.5 kat daha duyarsız olmuştur. Sonuç olarak, popülasyonların pestisitlere karşı duyarlılık farkları artan CarE, GST ve P-450 enzim aktivitelerini içeren çoklu fizyolojik mekanizmalarından kaynaklanmıştır. The susceptibilities to insecticides and acaricides, as well as carboxylesterase (CarE), cytochrome P450 monooxygenase (P450) and glutathione S-transferases (GSTs) activities were evaluated in a susceptible (SSS) and an unsusceptible strain (RSS) of Stethorus gilvifrons (Muls.) (Col.: Coccinellidae). Synergism studies using S, S, S-tributylphosphorotrithioate (DEF), piperonyl butoxide (PBO) and diethyl maleate (DEM) in combination with at a discrimination dose (similar to LD20) of selected pesticides were also conducted to test for possible physiological mechanisms. In addition, the insensitivity of acetylcholinesterase (AchE) to the inhibitors, paraoxon and pirimicarb, is also determined. Compared with SSS, tolerance ratios of RSS, as indicated by LC50 values, was calculated 2.1, 11.2, 6.7, 1.7, 1.6 and 2.3-fold, respectively, for abamectin, bifenazate, chlorpyrifos, cyhexatin, hexythiazox and lambda-cyhalothrin. Thus, the field collected resistant S. gilvifrons population were tolerant at medium level to four compounds. CarE activity was significantly higher in RSS than in SSS, 1.5-fold for alpha-naphthyl acetate and 4.3-fold for beta- naphthyl acetate. Also significant enhanced GSTs activity were determined in RSS using two substrates, 1-chloro-2,4-dinitrobenzene (4.3-fold) and 2,4-dinitro-1-iodobenzene (1.4-fold), compared with SSS. Inhibition kinetics revealed that AchE from RSS was 2.5-fold more insensitive to inhibition by 1mM primicarb than AchE from SSS. The disparity in susceptibility to these pesticides in the predator populations were due to the responsibility of multiple physiological mechanisms, including elevated CarE, GSTs and P-450 enzymes activities.

In 2002, typical symptoms of bacterial spot disease of stone fruits caused by Xanthomomas arboricola pv. pruni (Xap) were observed for the first time on Japanese plum in Badajoz (south-western Spain). During the following years, the pathogen was found in seven other eastern and northern Spanish provinces (Valencia, Alicante, Zaragoza, Huesca, Navarra, Lérida and Mallorca) affecting different cultivars of Japanese plum, nectarine, peach and almond. There are few previous reports of Xap on almond, the Spanish outbreaks constituting its first detection on this host in the European Union (EU). Identification of the pathogen was performed using biochemical tests, fatty acid methyl esters (FAME) profiles, conventional and real-time PCR, and hypersensitivity reaction on tobacco leaves. Pathogenicity was demonstrated by inoculation of young potted plants of peach, plum or almond and successful re-isolations from plants with symptoms. In areas where infected plants were found, eradication programs were set up since Xap has a quarantine status according to phytosanitary EU legislation. Published

Xanthomonas arboricola pv. pruni (Xap), the causal agent of bacterial spot disease of stone fruits and almond, is regulated as a quarantine pathogen in the European Union and the European and Mediterranean Plant Protection Organization (EPPO). Xap can have an epiphytic phase and/or be latent and, consequently, it can be transmitted by different types of plant material. Effective quarantine measures require specific, sensitive and rapid methods to detect Xap in propagative material or new reservoirs. Laborious and time-consuming methods for the diagnosis of Xap are recommended in the current EPPO standard protocol. However, new several pathogen-specific PCR and quantitative realtime PCR assays have been developed that enable direct detection of Xap in symptomatic and symptomless plant samples. A concise resource of current methods for Xap detection and identification, based on assessment and development activities within the framework of COST 873, is presented.

Introduction: Lipemic specimens are a common and frequent, but yet unresolved problem in clinical chemistry, and may produce significant interferences in the analytical results of different biochemical parameters. Material and methods: The aim of this study was to examine the effect of lipid removal using ultra-centrifugation of lipemic samples, on some routine biochemistry parameters. Among all the samples obtained daily in our laboratory, the ones which were visibly muddy were selected and underwent to a process of ultracentrifugation, being determined a variety of biochemical tests before and after ultra-centrifugation. A total of 110 samples were studied. Results: We found significant differences in all the parameters studied except for total bilirubin, glu-cose, gamma-glutamyl transferase (GGT) and aspartate aminotransferase (AST). The greatest diffe-rences in the parameters analyzed were found in the concentration of alanine aminotransferase (ALT) (7.36%) and the smallest ones in the concentration of glucose (0.014%). Clinically significant interfe-rences were found for phosphorus, creatinine, total protein and calcium. Conclusion: Lipemia causes clinically significant interferences for phosphorus, creatinine, total prote-in and calcium measurement and those interferences could be effectively removed by ultracentrifuga-tion.

A creatina é o suplemento nutricional mais popular utilizado para melhorar o desempenho em atividades que envolvem exercícios de curta duração e alta intensidade. Porém, as possíveis intercorrências advindas do seu uso não estão totalmente elucidadas. O objetivo do estudo foi avaliar os efeitos de duas dosagens de suplementação com creatina nas funções renal e hepática de adultos saudáveis durante oito semanas de treinamento de musculação. Exames bioquímicos foram realizados em 35 praticantes de musculação distribuídos aleatoriamente em três grupos, placebo (PLA, n = 12), creatina (CRE1, n = 12) e creatina 2 (CRE2, n = 11), antes e após oito semanas de treinamento com exercícios resistidos. Em desenho duplo-cego, os voluntários foram suplementados (20g/dia) com creatina (CRE1 e CRE2) ou placebo (PLA) por sete dias e nos 53 dias subsequentes com 0,03g/kg de massa corporal de creatina (CRE1) e placebo (PLA) e com 5g/dia o grupo CRE2. Não houve intervenção na composição de suas dietas, que foram registradas e analisadas. Os resultados dos exames bioquímicos realizados permaneceram dentro das faixas de normalidade. Os valores de creatinina aumentaram 12,2% no grupo CRE1 e 9,0%, no CRE2, enquanto que no grupo PLA diminuiu 4,7%, entretanto, esses valores não ultrapassaram os índices de normalidade. Os valores dos exames da função hepática diminuíram em quase todas as frações, em todos os tratamentos, contudo, sem significância estatística. Conclui-se que a suplementação com creatina nas dosagens utilizadas (0,03g/kg e 5g/dia) para indivíduos saudáveis por oito semanas não altera a função hepática ou renal, sendo assim, nas condições deste estudo, foi considerada segura. Creatine is the most popular nutritional supplement widely used to improve performance in activities that involve exercise of short duration and high intensity. However, the complications arising from its use are not fully elucidated. The aim of this study was to evaluate the effects of two doses of creatine supplementation on renal and hepatic function in healthy adults during eight weeks of resistance exercise training. Biochemical tests were performed on 35 athletes randomly distributed into three groups, placebo (PLA, n = 12), creatine (CRE1, n = 12) and creatine2 (CRE2, n = 11) before and after eight weeks of resistance training. In a double-blind design, the volunteers were supplemented (20 g/day) with creatine (CRE1, CRE2) or placebo (PLA) for seven days and at the 53 subsequent days with 0.03g/kg of body weight of each supplement (CRE1, PLA) and 5g/day for CRE2. There was no intervention in the composition of their diets, which were recorded and analyzed. The results of biochemical tests conducted remained within normal ranges. Creatinine values increased by 12.2% for CRE1 and 9.0% for CRE2, whereas decreased by 4.7% in PLA; however, these values did not exceed normal rates. The values of liver function tests declined in nearly all fractions in all treatments, not being statistically significant, though. It is concluded that creatine supplementation at the dosages used (0.03g/kg and 5g/day) for healthy subjects during eight weeks does not alter hepatic or renal function, hence under the conditions of this study, creatine was considered safe.

La escaldadura foliar (EF) causada por Xanthomonas albilineans (Ashby) Dowson, es una de las enfermedades de mayor importancia en el cultivo de la caña de azúcar, Saccharum sp., ocasionando pérdidas estimadas entre 90-100% de la producción. El uso de la reacción en cadena de la polimerasa (por sus siglas en inglés pcR) como herramienta de diagnóstico de fitopa tógenos se incrementó a nivel mundial dada su rapidez, sensi bilidad y especificidad. Dada la importancia que tiene esta enfermedad en Venezuela, se llevó a cabo el diagnóstico de la bacteria a partir de plantas de caña de azúcar sintomáticas, empleando pruebas microbiológicas, bioquí micas y moleculares. Utilizando diversos métodos de purifi cación se obtuvieron aislados bacterianos, los cuales se some tieron a análisis fenotípicos y bioquímicos, así como a ensayos de pcR con iniciadores espe cíficos y las secuencias consenso repetitivas intragénicas de entero bacterias (siglas en inglés ERIc-pcR). con el método de macerado de hojas se obtuvieron 50 aislados, de los cuales 14 se selec cionaron por pruebas microbio lógicas y bioquí micas, de estos, sólo tres cepas fueron identi ficadas como X. albilineans. Este resultado se corroboró al obtener por pcR la banda esperada de 360 pb con los iniciadores específicos sólo en estos tres aislados. Finalmente, con la técnica de genotipificación ERIC-PCR se obtuvieron patrones de bandas para las bacterias identificadas como X. albilineans que no son similares a los descritos por otros autores en otras regiones del mundo. Este trabajo representa la primera genotipificación de aislados de X. albilineans en Venezuela Leaf scald caused by Xanthomonas albilineans, is one of the most important diseases in sugarcane, Saccharum sp., which causes serious economical losses estimated between 90-100%. The use of pcR as a diagnostic tool for plant pathogens has increased because of its speed, sensitivity and specificity. Given the importance of leaf scald in Venezuela, in this work we carried out the diagnosis of X. albilineans from symptomatic sugarcane plants, using microbiological, biochemical and molecular methods. Bacterial isolates were obtained using different methods of purification. These isolates were then subjected to phenotypic and biochemical analysis, PCR with specific primers and ERIc pcR. With the method of macerated leaves 50 isolates were obtained, out of which 14 were selected for biochemical tests. From these 14 isolates only three strains were identified as X. albilineans by microbiological and biochemical tests. These results were confirmed by PCR by obtaining the expected band of 360 bp using specific primers. Finally, the ERIc-pcR banding patterns obtained for the isolates identified as X. albilineans were found to be different to those described by other authors in other regions. This work represents the first genotyping assay of isolates of X. albilineans in Venezuela

Evaluation of a simplified key for the identification of coagulase-positive Staphylococcus isolated from bovine mastitis. Three hundred fourty four strains of coagulase-positive Staphylococcus (CPS), isolated from mastitis cases, underwent phenotypic and genotypic tests to evaluate the efficiency of a simplified key, based on phenotypic tests for the discrimination of these microorganisms. The tests consisted of amplification of the femA gene and hemolysis in blood agar, production of acetoin and fermentation of maltose, mannitol and trehalose. Strains that showed negative results in the amplification test of the femA gene or that were not identified as Staphylococcus aureus (S. aureus) by phenotypic tests were tested with the APISTAPH kit (Biomériux-France), for precise identification of species. Phenotypic tests revealed 338 strains (98.25%) as S. aureus, three strains (0.86%) as Staphylococcus hyicus, and three microorganisms (0.86%) as Staphylococcus intermedius. PCR demonstrated that 338 (98.25%) strains belonged to the S. aureus species, confirming the results for 336 strains from 338 identified, through a simplified phenotypic key. A high rate of correlation (98.83%) was verifeid between the results of genotypic and phenotypic tests for the identification of S. aureus, demonstrating the applicability of the proposed key, for the discrimination of this microorganism in CPS isolated from bovine mastitis. Evaluation of a simplified key for the identification of coagulase-positive Staphylococcus isolated from bovine mastitis. Three hundred fourty four strains of coagulase-positive Staphylococcus (CPS), isolated from mastitis cases, underwent phenotypic and genotypic tests to evaluate the efficiency of a simplified key, based on phenotypic tests for the discrimination of these microorganisms. The tests consisted of amplification of the femA gene and hemolysis in blood agar, production of acetoin and fermentation of maltose, mannitol and trehalose. Strains that showed negative results in the amplification test of the femA gene or that were not identified as Staphylococcus aureus (S. aureus) by phenotypic tests were tested with the APISTAPH kit (Biomériux-France), for precise identification of species. Phenotypic tests revealed 338 strains (98.25%) as S. aureus, three strains (0.86%) as Staphylococcus hyicus, and three microorganisms (0.86%) as Staphylococcus intermedius. PCR demonstrated that 338 (98.25%) strains belonged to the S. aureus species, confirming the results for 336 strains from 338 identified, through a simplified phenotypic key. A high rate of correlation (98.83%) was verifeid between the results of genotypic and phenotypic tests for the identification of S. aureus, demonstrating the applicability of the proposed key, for the discrimination of this microorganism in CPS isolated from bovine mastitis.

Las infecciones causadas por micobacterias no tuberculosas (MNT) o atípicas constituyen en la actualidad un grave problema de salud, especialmente en pacientes inmunocomprometidos. Estas micobacterias presentan patrones de susceptibilidad a antibióticos particulares y distintos a M. tuberculosis, por lo que la administración del tratamiento adecuado requiere de un método rápido, sencillo y sensible de identificación. La técnica de PRA (Análisis de Restricción de Productos de PCR), basada en la digestión enzimática del producto de amplificación del gen hsp65, ha mostrado ser un método adecuado de identificación de micobacterias. En el presente trabajo se comparó la técnica de PRA con el estándar de identificación de micobacterias representado por las pruebas bioquímicas en 30 aislados provenientes del Laboratorio de Tuberculosis del Instituto de Biomedicina. La técnica de PRA permitió identificar 96% de las cepas analizadas, en comparación con 92.% de cepas identificadas por las técnicas bioquímicas. Los resultados obtenidos fueron idénticos en 18 de 22 cepas, correspondiendo al 82% de los resultados. Se concluye que el PRA es un método rápido, sencillo y económico que produce resultados concordantes con las técnicas tradicionales, con un menor grado de error. Basados en estos resultados se recomienda el uso del PRA en los laboratorios clínicos como método de identificación de rutina para micobacterias. Infections caused by atypical mycobacteria at present constitute a serious health problem, especially in immunocompromised patients. These mycobacteria present particular susceptibility patterns, different from M. tuberculosis, due to which the administration of an adequate treatment requires a fast, simple and sensitive identification method. The PRA technique (PCR Restriction), based on the enzymatic digestion of the amplification product of the hsp65 gene has shown to be an adequate method for the identification of mycobacteria. In this study we compared the PRA technique with the standard mycobacterial identification method, represented by biochemical tests, in 30 isolates from the Tuberculosis Laboratory of the Instituto de Biomedicina. The PRA technique allowed the identification of 96% of the strains analyzed, as compared with 92% of strains identified through biochemical methods. The results obtained were identical in 18 of 22 strains, corresponding to 82% of the results. It is concluded that the PRA technique is a fast, simple and economical method that produces results in concord with traditional techniques, with a lesser degree of error. Based in these results, the use of PRA as routine identification technique for mycobacteria is recommended for clinical laboratories.

No presente trabalho, avaliou-se o hemograma, diversas proteínas e enzimas séricas ou plasmáticas e a produção de anticorpos específicos em Felis domesticus, experimentalmente infectados por Lagochilascaris minor. Verificou-se nos animais infectados aumento de leucócitos totais, principalmente eosinófilos; queda do número de plaquetas; aumento de aspartato-aminotransferase e alanina-aminotransferase; e principalmente a presença de anticorpos IgG específicos para antígenos do parasita. A reação com extrato bruto de parasitas adultos mostrou-se mais específica, permitindo a discriminação de soros de animais: não infectados, com infecção por outros parasitas, e com lagochilascariose. Esta é a primeira descrição da padronização de uma reação sorológica para diagnóstico da lagochilascariose em Felis domesticus. The present study evaluated the hemogram, different proteins, plasma enzymes, serum enzymes and specific antibody production of Felis domesticus experimentally infected by Lagochilascaris minor. The infected animals were seen to present increased total leukocytes (particularly eosinophils), decreased platelet counts, increased aspartate-aminotransferase and alanine-aminotransferase and, especially, the presence of specific IgG antibodies against antigens of the parasite. The reaction with crude extract of adult parasites was shown to be more specific, thereby enabling serum discrimination between the animals: non-infected, infected with other parasites and infected with lagochilascariosis. This is the first description of the standardization of a serological reaction for diagnosing lagochilascariosis in Felis domesticus.

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T18:59:49Z No. of bitstreams: 1 S0103-84782007000200025.pdf: 123187 bytes, checksum: f2bef7e5ae9e93ac23a421ee5712fd17 (MD5) Made available in DSpace on 2013-08-22T18:59:49Z (GMT). No. of bitstreams: 1 S0103-84782007000200025.pdf: 123187 bytes, checksum: f2bef7e5ae9e93ac23a421ee5712fd17 (MD5) Previous issue date: 2007-04-01 Made available in DSpace on 2013-09-30T19:57:13Z (GMT). No. of bitstreams: 2 S0103-84782007000200025.pdf: 123187 bytes, checksum: f2bef7e5ae9e93ac23a421ee5712fd17 (MD5) S0103-84782007000200025.pdf.txt: 20332 bytes, checksum: 3b641496b9c8d4df5a5a269830274c5b (MD5) Previous issue date: 2007-04-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:15:56Z No. of bitstreams: 2 S0103-84782007000200025.pdf: 123187 bytes, checksum: f2bef7e5ae9e93ac23a421ee5712fd17 (MD5) S0103-84782007000200025.pdf.txt: 20332 bytes, checksum: 3b641496b9c8d4df5a5a269830274c5b (MD5) Made available in DSpace on 2014-05-20T13:15:56Z (GMT). No. of bitstreams: 2 S0103-84782007000200025.pdf: 123187 bytes, checksum: f2bef7e5ae9e93ac23a421ee5712fd17 (MD5) S0103-84782007000200025.pdf.txt: 20332 bytes, checksum: 3b641496b9c8d4df5a5a269830274c5b (MD5) Previous issue date: 2007-04-01 Um estudo sobre infecção experimental foi realizado em oito suínos, com idade média de 90 dias, machos castrados, da raça Wessex, e distribuídos em dois grupos de quatro suínos cada. Durante 36 dias, foram analisadas as alterações bioquímicas nos soros dos suínos dos dois grupos. O Grupo I foi mantido como testemunho e recebeu 5,0mL de solução fisiológica estéril por via intravenosa (veia cava craniana) e, no Grupo II, os suínos foram inoculados pela mesma via com 5,0mL de cultura de Leptospira interrogans sorovar wolffi , amostra L-10 selvagem isolada de tatu (Dasypus novemcinctus), contendo 1,0 x 10(8) leptospiras/mL. A partir do terceiro dia após a inoculação e em intervalos de 72 horas até o décimo oitavo dia, foram feitas coletas de sangue, sem anticoagulante, dos animais inoculados e testemunhas. Os parâmetros bioquímicos analisados foram: bilirrubina total, direta e indireta, ácidos graxos, glicose e proteínas plasmáticas. Foi detectado um aumento da bilirrubina direta no terceiro dia e um aumento no sexto dia da bilirrubina total e indireta após a inoculação. As dosagens de glicose, ácidos graxos e proteínas plasmáticas apresentaram uma diminuição a partir do terceiro dia da inoculação. Com os resultados obtidos, pode-se concluir que o aumento das taxas de bilirrubinas levam a uma definição de um diagnóstico de hemólise aguda, e que a hipoglicemia, a hipolipidemia e a hipoproteinemia podem estar relacionadas com lesões hepáticas e a uma septcemia.Todas as dosagens em todos os animais retornaram aos seus valores normais a partir do décimo quinto dia. Eight, 90 days old pigs, of the Wessex lineage all castrated male were used in experiment, divided into two groups of four animals. Biochemical alterations in the serum of the animals were analyzed in both groups during 36 days. Control (Group I) received 5.0mL of a 0.9% sterile sodium chloride solution by intracranial vein injection; Group II animals were inoculated by the same way with 5.0mL of a cell culture containing 1.0x10(8)cells/mL of Leptospira interrogans sorovar wolffi, wild strain L-10, isolated from a wild especies of brazilian armadillo (Dasypus novemcinctus). Three days after inoculation, blood was collected without anticoagulant; the same process was repeated at 72 hours intervals during eighteen days in both, control and experimental groups. Quantitative biochemical parameters were direct, indirect and total bilirubin, fatty acids and serum proteins. The experimental animals showed an increase of direct bilirubin three days after inoculation. Increases in indirect and total bilirubin were, also, observed after six days. Glucose, fatty acids and serum proteins decreased after the third day of inoculation. In conclusion, the increase in bilirubin levels could be due to acute hemolysis; hypoglycemia, hypolipidemia and hypoproteinemia that could be related to hepatic lesions and septicemia. All parameters returned to normal levels after fifteen days, in all animals tested. Universidade Estadual de São Paulo Faculdade de Ciências Agrárias e Veterinária Universidade de São Paulo Departamento de Morfologia, Estomatologia e Fisiologia Universidade Estadual de São Paulo Faculdade de Ciências Agrárias e Veterinária Departamento de Patologia Veterinária