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Sample records for animal mediante pcr

  1. Procesamiento de muestras fecales en el estudio de Cryptosporidium sp. mediante PCR

    Directory of Open Access Journals (Sweden)

    Gregorio Pérez-Cordón

    2013-05-01

    Full Text Available El presente trabajo describe un protocolo aplicado a muestras de heces de ganado bovino para el aislamiento y purificación de ooquistes de Cryptosporidium sp. y el eventual uso de los mismos para estudios genéticos mediante PCR.

  2. Detección proviral de HTLV-1 mediante reacción en cadena de la polimerasa (PCR)

    OpenAIRE

    Sevilla, Carlos; D.A. Microbiología Médica, Facultad de Medicina, UNMSM; Alarcón, Jorge; D.A. Microbiología Médica, Facultad de Medicina, UNMSM; Huiza, Alina; D.A. Microbiología Médica, Facultad de Medicina, UNMSM; Gutiérrez, César; D.A. Microbiología Médica, Facultad de Medicina, UNMSM; Ñavincopa, Marcos; D.A. Microbiología Médica, Facultad de Medicina, UNMSM; Cornejo, William; D.A. Microbiología Médica, Facultad de Medicina, UNMSM; Cáceres, Abraham; D. A. Microbiología Médica, Facultad de Medicina, UNMSM; Centro de Biotecnología, Universidad Iberoamericana de Ciencias y Tecnología; Laboratorio Entomología Médica, Sección Parasitología, Instituto de Salud Pública; Laboratorio de Entomología Médica, Laboratorio de Referencia Regional, Gerencia Regional de Salud La Libertad; Dirección Ejecutiva de Salud Ambiental, Sub Región de Salud “Luciano Castillo Colonna”, DIRESA Piura.; Tejada, Abelardo; D.A. Microbiología Médica, Facultad de Medicina, UNMSM; Romaní, Franco; D.A. Microbiología Médica, Facultad de Medicina, UNMSM; Mori, Nicanor; D.A. Microbiología Médica, Facultad de Medicina, UNMSM; Wong, Paolo; D.A. Microbiología Médica, Facultad de Medicina, UNMSM; Huamán, Víctor; D.A. Microbiología Médica, Facultad de Medicina, UNMSM; Vásquez, Rubén; D.A. Microbiología Médica, Facultad de Medicina, UNMSM; Tejada, Romina; D.A. Microbiología Médica, Facultad de Medicina, UNMSM; Flores, Alitza; D.A. Microbiología Médica, Facultad de Medicina, UNMSM

    2013-01-01

    Objetivos: Detectar el genoma proviral de HTLV-1 mediante el desarrollo de reacción en cadena de la polimerasa (PCR). Diseño: Descriptivo. Institución: D.A. Microbiología Médica, Facultad de Medicina, UNMSM. Participantes: Personas con y sin sospecha de HTLV-I. Principales medidas de resultados: detección de HTLV-1 mediante PCR. Resultados: El 71,4% de los pacientes con sospecha clínica de HTLV-I fue reactivo por métodos Inmunológico. Elisa HTLV I-II Biokit detectó 5 casos reactivos (X=2,359 ...

  3. Animal Species Identification by PCR – RFLP of Cytochrome b

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    Tomáš Minarovič

    2010-05-01

    Full Text Available An alternative DNA detection system is based on the polymerase chain reaction (PCR amplification of a segment of the mitochondrial cytochrome b gene. Subsequent cleavage by a restriction enzymes gives rise to a specie-specific pattern on an agarose gel. We used five animal species (Mustela vison, Mustela putorius furo, Sus scrofa domesticus, Oryctolagus cuninculus, Anser anser. Length of PCR product was 359 bp and we used universal primers. Restriction fragment length polymorphism was analyzed by using the restriction endonuclease AluI. Results of cleavage were visualized by using electrophoresis and UV transiluminator. Every animal specie has a unique combination of restriction fragments i.e. Mustela vison 81 bp, 109 bp and 169 bp, Mustela putorius furo 169 bp and 190 bp, Sus scrofa domesticus 115 bp and 244 bp, Oryctolagus cunninculus is not cleaved by AluI so it has whole 359 bp fragment on agarose gel, Anser anser 130 bp and 229 bp. The results suggest that the method of PCR - RFLP is rapid and simple method for identification of species. PCR – RFLP can reliably identify chosen species. Application of genetic methods is very useful for breeding of livestock and protection of biodiversity.

  4. Detección rápida de resistencia a drogas en Mycobacterium tuberculosis mediante PCR-SSCP y PCR- Heteroduplex

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    Róger Calderón E

    2003-04-01

    Full Text Available Objetivo: Detectar tempranamente la susceptibilidad a las drogas antituberculosas rifampicina e isoniacida mediante PCR y electroforesis conformacional. Materiales y métodos: Se implementaron dos ensayos de amplificación de los genes rpoB y katG y mediante Heteroduplex y SSCP se determinó la susceptibilidad antituberculosa de 31 muestras clínicas procedentes de pacientes con diagnóstico de tuberculosis pulmonar baciloscopía positiva. La caracterización fenotípica de la susceptibilidad, se realizó empleando el método de las proporciones. Resultados: Los ensayos de PCR detectaron hasta 2,5 pg de ADN genómico de M. tuberculosis; no amplificando ADN de otras micobacterias y bacterias comunes de la flora bucal. Se encontró una concordancia general entre la detección molecular y convencional de la susceptibilidad a rifampicina e isoniacida de 96,7% y 83,9% (p<0,05, respectivamente. Sin embargo, sólo en pacientes con antecedente de tratamiento se presentó una concordancia del 100% y 90,9% (p<0,05 para rifampicina e isoniacida, respectivamente. Además, este sistema de detección de resistencia puede emitir resultados 48 horas después de la recepción de la muestra clínica. Conclusiones: Estos sistemas se presentan como una excelente alternativa para la identificación temprana de pacientes infectados con bacilos de M. tuberculosis drogoresistentes. Potencialmente, se podrán dirigir óptimos y oportunos esquemas terapéuticos que contribuirán con el control y prevención de la transmisión de cepas multidrogo-resistentes que afectan en gran medida a la salud pública de nuestro país.

  5. Detección del virus de la leucosis bovina en ganado criollo colombiano mediante PCR-anidado

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    Darwin Yovanny Hernández-Herrera

    2011-12-01

    Full Text Available Se evaluó la presencia del virus de la leucosis bovina (VLB en 360 muestras de ADN de ocho razas bovinas criollas: Blanco Orejinegro (BON, Casanareño (CAS, Costeño con Cuernos (CCC, Chino Santandereano (ChS, Caqueteño (CQT, Hartón del Valle (HV, Romosinuano (RS y San Martinero (SM, dos Razas Sintéticas Colombianas: Lucerna (LUC y Velásquez (VEL y dos razas foráneas: Brahmán (B y Holstein (H. Para la detección del pro-virus se amplificó una región del gen env viral, mediante PCR anidada. La presencia del VLB fue mayor en la raza HV seguido por ChS (83.3% y 60% respectivamente, VEL y LUC tuvieron el mismo porcentaje (50%, en CAS, CCC y CQT la presencia del virus fue de 26.7%, 23.3% y 16.7% respectivamente; no se encontró el virus en BON, SM y RS. En las razas foráneas la presencia fue de 83.3% para H y 6.7% para B. Se encontró dependencia altamente significativa entre la presencia del VLB y la raza, el sexo y región de origen de la muestra. El promedio de presencia en las razas criollas fue menor que en las foráneas, menor en los machos que en las hembras y en la región norte que en el suroccidente y el centro del país.

  6. Clostridium difficile PCR Ribotypes from Different Animal Hosts and Different Geographic Regions

    DEFF Research Database (Denmark)

    Zidaric, V.; Janezic, S.; Indra, A.

    Clostridium difficile is an anaerobic sporogenic bacterium traditionally associated with human nosocomial infections, and animals have been recognized as an important potential reservoir for human infections (Rodriguez-Palacios et al., 2013). Ribotype 078 is often reported in animals but accordin...... was to establish an international C. difficile animal collection with one PCR ribotype per species per country/laboratory and to compare PCR ribotypes across animal hosts and countries....... to recent studies the overlap between PCR ribotypes found in humans and animals seems to be increasing (Bakker et al., 2010; Gould and Limbago, 2010; Janezic et al., 2012; Keel et al., 2007; Koene et al., 2011). However, genetic diversity among animal strains remains poorly understood. The aim of our work...

  7. Quantitative polymerase chain reaction (PCR) for detection of aquatic animal pathogens in a diagnostic laboratory setting

    Science.gov (United States)

    Purcell, Maureen K.; Getchell, Rodman G.; McClure, Carol A.; Weber, S.E.; Garver, Kyle A.

    2011-01-01

    Real-time, or quantitative, polymerase chain reaction (qPCR) is quickly supplanting other molecular methods for detecting the nucleic acids of human and other animal pathogens owing to the speed and robustness of the technology. As the aquatic animal health community moves toward implementing national diagnostic testing schemes, it will need to evaluate how qPCR technology should be employed. This review outlines the basic principles of qPCR technology, considerations for assay development, standards and controls, assay performance, diagnostic validation, implementation in the diagnostic laboratory, and quality assurance and control measures. These factors are fundamental for ensuring the validity of qPCR assay results obtained in the diagnostic laboratory setting.

  8. Identificazione rapida di mutazioni associate a farmaco-resistenza in ceppi di citomegalovirus umano mediante nPCR-RFLP

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    Maria Cristina Medici

    2007-03-01

    Full Text Available We developed a nested-PCR followed by restriction fragment length polymorphism (RFLP for the detection of human cytomegalovirus (HCMV UL97 M460V/I, H520Q, C592Q,A594V, L595S/F and C603W mutations associated to ganciclovir (GCV resistance.The method uses five primer pairs and seven enzymes already published and newly combined.The detection limit of nPCR was assessed in a single serial dilution assay to be about 0.13 PFU/reaction. Expected restriction fragment patterns were obtained by nPCR-RFLP on either wild-type reference strains or strains and sequences of HCMV containing mutations. Then the nPCR-RFLP was used on 24 sera/plasma belonging to 22 transplant recipients (kidney, bone marrow, or kidney-pancreas: 13 subjects never treated with GCV (control group and 9 subjects treated with GCV oral profilaxis (study group. All codons detected from the control group (six in 8 cases and four in 1 case were identified as wild-type. All codons detected from the study group (six in 6 cases, three in 2 cases, and four in the second sample of 1 case whose first sample was negative by nPCR were wild-type except one, which showed a restriction pattern referring to M460V and/or M460I ATA-codified, definitively proved to be M460V by sequence analysis.This was the case of a renal transplant recipient at the end of profilaxis. In conclusion, the procedure seems to be quite sensitive and specific as well as able to detect mixed population of mutants or mutants and wild-type. It could represent a good tool in monitoring the emergence of HCMV mutants in renal transplant recipients treated with GCV.

  9. Detection of wild animals as carriers of Leptospira by PCR in the Pantanal biome, Brazil.

    Science.gov (United States)

    Vieira, Anahi S; Narduche, Lorena; Martins, Gabriel; Schabib Péres, Igor A H F; Zimmermann, Namor P; Juliano, Raquel S; Pellegrin, Aiesca O; Lilenbaum, Walter

    2016-11-01

    Leptospiral infection is widespread in wildlife. In this context, wild ecosystems in tropical countries hold a vast biodiversity, including several species that may act as potential reservoirs of leptospires. The Pantanal biome presents highly favorable environmental conditions for the occurrence of leptospirosis, such as high temperatures, constant flooding, and high biodiversity. The purpose of this study was to detect wild animals as carriers of Leptospira sp. using direct methods (PCR and culture) in the Pantanal biome, Brazil. A total of 35 animals were studied, namely Cerdocyon thous, Nasua nasua, Ozotoceros bezoarticus, and Sus scrofa species. Blood for serology (MAT) and urine for bacteriological culturing and PCR was sampled. The most prevalent serogroups were Javanica and Djasiman. Additionally, 40.6% of these animals presented PCR positive reactions. Seroreactivity associated with the high frequency of leptospiral carriers among the different studied species suggests a high level of exposure of the studied animals to pathogenic Leptospira strains. Our results are still limited and the actual role of the studied animals in the epidemiology of leptospirosis in the Pantanal region remains to be elucidated.

  10. IDENTIFICACIÓN MEDIANTE PCR DEL SEXO DE LA PAPAYA (Carica papaya L., HÍBRIDO "POCOCÍ"

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    Ericka Saalau-Rojas

    2009-01-01

    con dos metodologías de extracción, CTAB y lisis alcalina (NaOH. La amplificación por PCR del ADN extraído de muestras foliares de papaya híbrido "Pococí", con ambos métodos de extracción, produjo los fragmentos del tamaño esperado. La determinación del sexo de 1.500 plántulas en almácigo mostró un 46 % de plántulas hermafroditas y un 54 % de plantas femeninas. La proporción observada de plantas femeninas: hemafroditas no varió de la esperada (1:1 según la prueba de chi-cuadrado (p= 0,4237. Las plantas hermafroditas fueron llevadas al campo y al momento de la floración se determinó su sexo. La correspondencia entre el sexado por PCR y la expresión sexual en campo fue de un 98 %.

  11. REAL-TIME PCR DETECTION OF LISTERIA MONOCYTOGENES IN FOOD SAMPLES OF ANIMAL ORIGIN

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    Jaroslav Pochop

    2013-02-01

    Full Text Available The aim of this study was to follow the contamination of food with Listeria monocytogenes by using Step One real time polymerase chain reaction (PCR. We used the PrepSEQ Rapid Spin Sample Preparation Kit for isolation of DNA and SensiFAST SYBR Hi-ROX Kit for the real-time PCR performance. In 24 samples of food of animal origin without incubation were detected strains of Listeria monocytogenes in 15 samples (swabs. Nine samples were negative. Our results indicated that the real-time PCR assay developed in this study could sensitively detect Listeria monocytogenes in food of animal origin without incubation. This could prevent infection caused by Listeria monocytogenes, and also could benefit food manufacturing companies by extending their product’s shelf-life as well as saving the cost of warehousing their food products while awaiting pathogen testing results. The rapid real-time PCR-based method performed very well compared to the conventional method. It is a fast, simple, specific and sensitive way to detect nucleic acids, which could be used in clinical diagnostic tests in the future.

  12. Validation of qPCR Methods for the Detection of Mycobacterium in New World Animal Reservoirs.

    OpenAIRE

    Genevieve Housman; Joanna Malukiewicz; Vanner Boere; Grativol, Adriana D.; Pereira, Luiz Cezar M.; Ita de Oliveira Silva; Carlos R. Ruiz-Miranda; Richard Truman; Anne C Stone

    2015-01-01

    Zoonotic pathogens that cause leprosy (Mycobacterium leprae) and tuberculosis (Mycobacterium tuberculosis complex, MTBC) continue to impact modern human populations. Therefore, methods able to survey mycobacterial infection in potential animal hosts are necessary for proper evaluation of human exposure threats. Here we tested for mycobacterial-specific single- and multi-copy loci using qPCR. In a trial study in which armadillos were artificially infected with M. leprae, these techniques were ...

  13. Detección y cuantificación del Potato mop-top virus (PMTV en Colombia mediante qRT-PCR

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    Nevar García Bastidas

    2013-04-01

    Full Text Available El Potato mop-top virus (PMTV es uno de los virus re-emergentes en cultivos de papa en Colombia. Es transmitido por Spongospora subterranea, el agente causal de la sarna polvosa. La detección del PMTV presenta dificultades debido a su distribución irregular en las plantas, bajo título y movimiento sistémico como ARN desnudo. Con el fin de ampliar el rango de herramientas disponibles para detectar el PMTV en los programas de certificación de tubérculo-semilla, en este estudio se evaluó la prueba de RT-PCR en tiempo real (qRT-PCR en dos pasos: con los cebadores PMTV-1948F/PMTV-2017R y la sonda Taqman® PMTV-1970, dirigidos al gen CP-RT del ARN2 viral. Se construyó una curva estándar a partir de la transcripción in vitro de un fragmento de 1513 pb de este gen. Posteriormente, se evaluó la utilidad de la técnica a partir de tres tipos de muestras: plantas señuelo de Nicotiana benthamiana y Solanum phureja inoculadas con quistosoros de Sss, raíces de papa con síntomas de sarna polvosa del municipio de La Unión (Antioquia y tubérculos-semilla. Mediante qRT-PCR fue posible detectar el virus en 11 de las 20 muestras de raíz de plantas señuelo, mientras que 14 de las 15 muestras de raíces de papa resultaron positivas, estimándose una concentración entre 4.72 x 10(11 y 7.60 x 10(13 partículas virales/µl. Adicionalmente, en el ensayo de tubérculo-semilla se determinó la presencia del PMTV en una de las 16 muestras. Estos resultados indican la viabilidad de utilizar rutinariamente la técnica de qRT-PCR para la detección de PMTV en Colombia.

  14. Validation of qPCR Methods for the Detection of Mycobacterium in New World Animal Reservoirs.

    Directory of Open Access Journals (Sweden)

    Genevieve Housman

    2015-11-01

    Full Text Available Zoonotic pathogens that cause leprosy (Mycobacterium leprae and tuberculosis (Mycobacterium tuberculosis complex, MTBC continue to impact modern human populations. Therefore, methods able to survey mycobacterial infection in potential animal hosts are necessary for proper evaluation of human exposure threats. Here we tested for mycobacterial-specific single- and multi-copy loci using qPCR. In a trial study in which armadillos were artificially infected with M. leprae, these techniques were specific and sensitive to pathogen detection, while more traditional ELISAs were only specific. These assays were then employed in a case study to detect M. leprae as well as MTBC in wild marmosets. All marmosets were negative for M. leprae DNA, but 14 were positive for the mycobacterial rpoB gene assay. Targeted capture and sequencing of rpoB and other MTBC genes validated the presence of mycobacterial DNA in these samples and revealed that qPCR is useful for identifying mycobacterial-infected animal hosts.

  15. Identification of trypanosomes in wild animals from Southern Cameroon using the polymerase chain reaction (PCR

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    Herder S.

    2002-12-01

    Full Text Available One possible explanation of the maintenance of many historical foci of sleeping sickness in Central Africa could be the existence of a wild animal reservoir. In this study, PCR was used to detect the different trypanosome species present in wild animal captured by hunters in the southern forest belt of Cameroon (Bipindi. Trypanosomes were also detected by a parasitological method (Quantitative buffy coat : QBC. Parasite could not be isolated in culture medium (Kit for in vitro isolation : KIVI. Specific primers of T. brucei s.l., T. congolense forest type, T. congolense savannah type, T. vivax, T. simiae and T. b. gambiense group 1 were used to identify parasites in the blood of 164 animals belonging to 24 different species including ungulates, rodents, pangolins, carnivores, reptiles and primates. Of the 24 studied species, eight were carrying T. b. gambiense group 1. Those parasites pathogenic to man were found in monkeys (Cercocebus torquatus and Cercopithecus nictitans, in ungulates (Cephalophus dorsalis and C. monticola, in carnivores (Nandinia binotata and Genetta servalina and in rodents (Cricetomys gambianus and Atherurus africanus. 13 species (54 % were carrying T. brucei s.l. identified as non-gambiense group 1.

  16. Polimorfismo genético de beta-lactoglobulina y alphalactoalbúmina en el ganado criollo colombiano, mediante PCR-SSCP

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    Jaime A Rosero-Alpala

    2011-12-01

    Full Text Available La población de ganado criollo colombiano ha venido presentando una inquietante disminución al pasar de 23.415 ejemplares en 1999 a 20.102 en 2003. A pesar de los esfuerzos por recuperar las razas criollas el panorama para su conservación es incierto, por tanto la búsqueda de caracteres deseables puede contribuir a su valoración y conservación. Los genes relacionados con el mejoramiento de la calidad de la leche producida por estas razas se consideran de gran importancia en la industria láctea, por tal razón y con el objetivo de caracterizar los genes beta-lactoglobulina y alpha-lactoalbúmina se analizaron 30 muestras de sangre de cada una de las razas criollas (Blanco Orejinegro, Caqueteño, Casanareño, Costeño con cuernos, Chino Santandereano, Hartón del Valle, Romosinuano y Sanmartinero, dos razas sintéticas colombianas (Lucerna y Velásquez y dos razas foráneas (Holstein y Brahman. Se amplificaron fragmentos de 262pb para beta-lactoglobulina (b-LG y de 166 pb para alpha-lactoalbúmina (a-LA que se genotipificaron mediante PCR-SSCP. El promedio de la frecuencia para b-LG A y b-LG B fue de 0.46 ± 0.020 y de 0.53 ± 0.020, respectivamente, y de 0.35 ± 0.019 para a-LA A y 0.64 ± 0.019 para a-LA B. El promedio de diversidad genética (He para b-LG fue 0.498 y de 0.455 para a-LA. Los ganados criollos representan una base genética valiosa, como alternativa para mejorar genéticamente los hatos destinados a la producción de leche con mejores características en calidad para la industria láctea.

  17. Polimorfismo genético de beta-lactoglobulina y alphalactoalbúmina en el ganado criollo colombiano, mediante PCR-SSCP

    Directory of Open Access Journals (Sweden)

    Muñoz Florez Jaime Eduardo

    2011-12-01

    Full Text Available La población de ganado criollo colombiano ha venido presentando una inquietante disminución al pasar de 23.415 ejemplares en 1999 a 20.102 en 2003. A pesar de los esfuerzos por recuperar las razas criollas el panorama para su conservación es incierto, por tanto la búsqueda de caracteres deseables puede contribuir a su valoración y conservación. Los genes relacionados con el mejoramiento de la calidad de la leche producida por estas razas se consideran de gran importancia en la industria láctea, por tal razón y con el objetivo de caracterizar los genes beta-lactoglobulina y alpha-lactoalbúmina se analizaron 30 muestras de sangre de cada una de las razas criollas (Blanco Orejinegro, Caqueteño, Casanareño, Costeño con cuernos, Chino Santandereano, Hartón del Valle, Romosinuano y Sanmartinero, dos razas sintéticas colombianas (Lucerna y Velásquez y dos razas foráneas (Holstein y Brahman. Se amplificaron fragmentos de 262pb para beta-lactoglobulina (b-LG y de 166 pb para alpha-lactoalbúmina (a-LA que se genotipificaron mediante PCR-SSCP. El promedio de la frecuencia para b-LG A y b-LG B fue de 0.46 ± 0.020 y de 0.53 ± 0.020, respectivamente, y de 0.35 ± 0.019 para a-LA A y 0.64 ± 0.019 para a-LA B. El promedio de diversidad genética (He para b-LG fue 0.498 y de 0.455 para a-LA. Los ganados criollos representan una base genética valiosa, como alternativa para mejorar genéticamente los hatos destinados a la producción de leche con mejores características en calidad para la industria láctea.

  18. Detection of rendered meat and bone meals by PCR is dependent on animal species of origin and DNA extraction method.

    Science.gov (United States)

    Myers, Michael J; Farrell, Dorothy E; Deaver, Christine M; Mason, Jacquline; Swaim, Heidi L; Yancy, Haile F

    2010-06-01

    The capability of eight commercially available DNA extraction kits to extract bovine DNA originating in meat and bone meal from fortified feed was evaluated. Four different batches of bovine meat and bone meal (BMBM) were used for DNA extraction with the eight commercial DNA extraction kits. Within each kit, there were minimal differences in the batch-to-batch amounts of extracted DNA. There were differences between the kits in the amounts of DNA that could be extracted from the same amount of starting BMBM. These differences did not translate into differences in the amount of amplifiable DNA from BMBM-fortified dairy feed. Using a validated real-time PCR method, the kit yielding the highest amount extractable DNA was completely unable to yield a positive PCR result; one other kit was also unable to produce a positive PCR result from DNA extracted from BMBM-fortified feed. There was a complete lack of a correlation between the amount of bovine DNA isolated from BMBM by a given extraction kit compared with the relative amounts of DNA isolated from fortified animal feed as evidenced by the cycle threshold values generated using the real-time PCR method. These results demonstrate that extraction of DNA from processed animal protein is different for pure ingredients and fortified animal feeds. These results indicate that a method specifically developed using just animal-derived meat and bone meal may not yield a functional assay when used to detect animal tissues in complete animal feed.

  19. Polimorfismo genético de beta-lactoglobulina y alphalactoalbúmina en el ganado criollo colombiano, mediante PCR-SSCP Genetic polymorphism of beta-lactoglobulin and alpha-lactoalbumin in Colombian Creole cattle by PCR-SSCP

    Directory of Open Access Journals (Sweden)

    Jaime A Rosero-Alpala

    2011-12-01

    Full Text Available La población de ganado criollo colombiano ha venido presentando una inquietante disminución al pasar de 23.415 ejemplares en 1999 a 20.102 en 2003. A pesar de los esfuerzos por recuperar las razas criollas el panorama para su conservación es incierto, por tanto la búsqueda de caracteres deseables puede contribuir a su valoración y conservación. Los genes relacionados con el mejoramiento de la calidad de la leche producida por estas razas se consideran de gran importancia en la industria láctea, por tal razón y con el objetivo de caracterizar los genes beta-lactoglobulina y alpha-lactoalbúmina se analizaron 30 muestras de sangre de cada una de las razas criollas (Blanco Orejinegro, Caqueteño, Casanareño, Costeño con cuernos, Chino Santandereano, Hartón del Valle, Romosinuano y Sanmartinero, dos razas sintéticas colombianas (Lucerna y Velásquez y dos razas foráneas (Holstein y Brahman. Se amplificaron fragmentos de 262pb para beta-lactoglobulina (b-LG y de 166 pb para alpha-lactoalbúmina (a-LA que se genotipificaron mediante PCR-SSCP. El promedio de la frecuencia para b-LG A y b-LG B fue de 0.46 ± 0.020 y de 0.53 ± 0.020, respectivamente, y de 0.35 ± 0.019 para a-LA A y 0.64 ± 0.019 para a-LA B. El promedio de diversidad genética (He para b-LG fue 0.498 y de 0.455 para a-LA. Los ganados criollos representan una base genética valiosa, como alternativa para mejorar genéticamente los hatos destinados a la producción de leche con mejores características en calidad para la industria láctea.The Colombian Creole Cattle has showed a preoccupant population decreasing, from 23,415 individuals in 1999 to 20,102 in 2003. Despite that many efforts to recover the creole breeds have been done, its future conservation is unclear. Searching for economic desirable genes may contribute to its preservation and utilization as a genetic resource. Genes related with the improvement of milk proteins are considered as an economic important

  20. Detección del virus de la leucosis bovina en ganado criollo colombiano mediante PCR-anidado Bovine leukemia virus detection in Creole Colombian breeds using nested-PCR

    Directory of Open Access Journals (Sweden)

    Darwin Yovanny Hernández-Herrera

    2011-12-01

    Full Text Available Se evaluó la presencia del virus de la leucosis bovina (VLB en 360 muestras de ADN de ocho razas bovinas criollas: Blanco Orejinegro (BON, Casanareño (CAS, Costeño con Cuernos (CCC, Chino Santandereano (ChS, Caqueteño (CQT, Hartón del Valle (HV, Romosinuano (RS y San Martinero (SM, dos Razas Sintéticas Colombianas: Lucerna (LUC y Velásquez (VEL y dos razas foráneas: Brahmán (B y Holstein (H. Para la detección del pro-virus se amplificó una región del gen env viral, mediante PCR anidada. La presencia del VLB fue mayor en la raza HV seguido por ChS (83.3% y 60% respectivamente, VEL y LUC tuvieron el mismo porcentaje (50%, en CAS, CCC y CQT la presencia del virus fue de 26.7%, 23.3% y 16.7% respectivamente; no se encontró el virus en BON, SM y RS. En las razas foráneas la presencia fue de 83.3% para H y 6.7% para B. Se encontró dependencia altamente significativa entre la presencia del VLB y la raza, el sexo y región de origen de la muestra. El promedio de presencia en las razas criollas fue menor que en las foráneas, menor en los machos que en las hembras y en la región norte que en el suroccidente y el centro del país.Using 360 DNA samples from eight Creole bovine breeds Blanco Orejinegro (BON, Casanareño (CAS, Costeño con Cuernos (CCC, Chino Santandereano (ChS, Caqueteño (CQT, Hartón del Valle (HV, Romosinuano (RS and San Martinero (SM, two synthetic Colombian breeds: Lucerna (LUC and Velásquez (VEL and two introduced breeds Brahmán (B and Holstein (H; the presence of Bovine Leukemia Virus (BLV was evaluated through the amplification of a viral gene region env (provirus detection - nested-PCR. The percentage of presence and independence test were calculated (X². Presence of BLV was higher in HV breed, followed by ChS (83.3% and 60% respectively; VEL and LUC breeds showed the same percentage (50%. In CAS, CCC and CQT the presence of virus was 26.7%, 23.3% y 16.7% respectively. On the other hand, no virus presence was

  1. Análisis resistivo de un nuevo arado de tracción animal mediante el Método de Elementos Finitos (MEF

    Directory of Open Access Journals (Sweden)

    Fidel Diego Nava

    2012-01-01

    Full Text Available La tracción animal como fuente energética tiene una amplia utilización en la agricultura de Oaxaca, México. Por tal razón se desarrolló un nuevo arado de tracción animal para realizar las distintas operaciones que se requieren durante la labranza. Con fines de la optimización se hizo necesario analizar la resistencia de los elementos estructurales de dicho arado mediante el método de elementos finitos. Como primer paso se desarrolló el modelo geométrico con la herramienta computacional Cosmos DesingStar 2008, y la definición de las propiedades mecánicas de los materiales empleados en la construcción del arado, así como la definición de las condiciones de borde y cargas. Los resultados mostraron que la estructura resiste las cargas aplicadas durante el trabajo con un coeficiente de seguridad de 2,46, la magnitud de las deformaciones resultantes se limitaron a 0,0005 m. Se evidencia que los materiales empleados en la construcción del arado garantizan la resistencia y rigidez requerida para el trabajo sin fallos de sus elementos y órganos de trabajo.

  2. Evaluation of immunomagnetic separation and PCR for the detection of Escherichia coli O157 in animal feces and meats

    NARCIS (Netherlands)

    Islam, M.A.; Heuvelink, A.E.; Talukder, K.A.; Zwietering, M.H.; Boer, de E.

    2006-01-01

    Series of animal feces and meat samples artificially contaminated with strains of Escherichia coli O157 isolated from different sources were tested by both an immunomagnetic separation (IMS)-based method and a PCR method using primers specific for a portion of the rfbE gene of E. coli O157. IMS is l

  3. A novel PCR-based method to enumerate Salmonella in animal feed

    DEFF Research Database (Denmark)

    Löfström, Charlotta; Andersson, Gunnar; Häggblom, Per

    2010-01-01

    the pellet and subjected to real-time PCR. The qualitative PCR method was compared to a reference culture method using modified semisolid Rappaport-Vassilades (MSRV) agar plates (ISO 6579, Amd D, 2007). Of 81 naturally or artificially contaminated samples tested (soya meal, rape seed meal, rape seed cake...

  4. Development and evaluation of an ITS1 "Touchdown" PCR for assessment of drug efficacy against animal African trypanosomosis.

    Science.gov (United States)

    Tran, Thao; Napier, Grant; Rowan, Tim; Cordel, Claudia; Labuschagne, Michel; Delespaux, Vincent; Van Reet, Nick; Erasmus, Heidi; Joubert, Annesca; Büscher, Philippe

    2014-05-28

    Animal African trypanosomoses (AAT) are caused by flagellated protozoa of the Trypanosoma genus and contribute to considerable losses in animal production in Africa, Latin America and South East Asia. Trypanosoma congolense is considered the economically most important species. Drug resistant T. congolense strains present a threat to the control of AAT and have triggered research into discovery of novel trypanocides. In vivo assessment of trypanocidal efficacy relies on monitoring of treated animals with microscopic parasite detection methods. Since these methods have poor sensitivity, follow-up for up to 100 days after treatment is recommended to increase the chance of detecting recurrent parasitaemia waves. Molecular techniques are more amendable to high throughput processing and are generally more sensitive than microscopic detection, thus bearing the potential of shortening the 100-day follow up period. The study presents a "Touchdown" PCR targeting the internal transcribed spacer 1 of the ribosomal DNA (ITS1 TD PCR) that enables detection and discrimination of different Trypanosoma taxa in a single run due to variations in PCR product sizes. The assay achieves analytical sensitivity of 10 parasites per ml of blood for detection of T. congolense savannah type and T. brucei, and 100 parasites per ml of blood for detection of T. vivax in infected mouse blood. The ITS1 TD PCR was evaluated on cattle experimentally infected with T. congolense during an investigational new veterinary trypanocide drug efficacy study. ITS1 TD PCR demonstrated comparable performance to microscopy in verifying trypanocide treatment success, in which parasite DNA became undetectable in cured animals within two days post-treatment. ITS1 TD PCR detected parasite recrudescence three days earlier than microscopy and had a higher positivity rate than microscopy (84.85% versus 57.58%) in 66 specimens of relapsing animals collected after treatments. Therefore, ITS1 TD PCR provides a useful tool

  5. Genotyping of Hydatid Cyst Isolated from Human and Domestic Animals in Ilam Province, Western Iran Using PCR-RFLP

    Directory of Open Access Journals (Sweden)

    M Dousti

    2013-03-01

    Full Text Available Background: Hydatidosis or cystic hydatid disease is one of the most important diseases in human and animals. Identification of strains is important for improvement of control and prevention of dis­ease. The aim of this study was to determine the strains isolated from human and domestic animals in Ilam Province, Iran, using PCR-RFLP method.Methods: Respectively, 30 and 4 animal and human hydatid cysts were collected from different slaughter­houses and hospitals of the province. Protoscolices were separated and their DNA genome was extracted by extraction kit. rDNA-ITS1 of each isolated samples was duplicated by BD1(Forward and 4s (Reverse Primers. PCR products were studied by electrophoresis and then were digested using TaqI, HpaII, RsaI and AluI restriction enzymes. RFLP products were studied using electrophoresis on 1% agar gel.Result: A fragment of 1000bp was produced from amplification of rDNA-ITS1 of protoscolices using PCR method. After digestion of PCR product by AluI enzyme, 200bp and 800bp, by RsaI, 655bp and 345bp and by HpaII 700bp and 300bp sizes were obtained. TaqI enzyme had no change in fragment size and it remained 1000bp. Considering the method, Ilam strains was specified as E. granulosus sensu stricto (G1-G3.Conclusions: Although sheep strain (G1 is dominated in human and different animal in Iran and the world, but more efforts should be done to clarify the true genotype of Ilam strains specified as E. granulosus sensu stricto (G1-G3.

  6. Detection of Brucella melitensis DNA in the milk of sheep after abortion by PCR assay Detección de ADN de Brucella melitensis mediante la prueba de PCR en muestras de leche de ovejas postaborto

    Directory of Open Access Journals (Sweden)

    Z Ilhan

    2008-01-01

    Full Text Available Laboratory diagnosis of brucellosis is generally performed by microbiological and serological methods. PCR assay is a specific and sensitive choice for the detection of different bacterial agents. An evaluation of this test was carried out for the detection of Brucella melitensis DNA in sheep milk. 102 milk samples from sheep after abortion were taken and studied using bacteriological culture, PCR and milk ring test (MRT. PCR found B. melitensis DNA in 24 (23.5% out of 102 milk samples, while only 8 (7.8% of the samples were positive to B. melitensis through direct culture. MRT found 28 (27.4% positive milk samples. The detection limit for PCR in sheep milk inoculated with B. melitensis strain 16 M was 1.7x10³-1.7x10(4 cfu/ml. PCR and MRT coincidence was 96%. The diagnostic sensitivity and specificity were determined as 100% and 81.3% respectively for PCR assay and 75% and 75% for MRT. PCR is a useful tool for a fast diagnosis of B. melitensis in sheep milk.El diagnóstico de laboratorio de brucelosis es generalmente realizado por métodos microbiológicos y serológicos. La prueba PCR es reconocida como una alternativa específica y sensible para la detección de diferentes agentes bacterianos. Se realizó una evaluación de la prueba PCR para la detección de ADN de Brucella melitensis en leche de oveja. Ciento dos muestras tomadas de ovejas postaborto fueron analizadas por métodos de cultivo bacteriológico, prueba PCR y prueba del anillo en leche (MRT. El PCR detectó ADN de B. melitensis en 24 (23,5% de 102 muestras de leche, mientras que solamente 8 (7,8% muestras de leche fueron positivas a B. melitensis por cultivo directo. El MRT detectó 28 (27,4% muestras positivas de leche. El límite de detección de B. melitensis 16 M por PCR fue de 1,7x10³-1,7x10(4 ufc/ml en leche inoculada. La concordancia entre PCR y MRT fue 96%. La sensibilidad diagnóstica y la precisión fueron determinadas como el 100% y el 81,3% respectivamente para la

  7. Avian-specific real-time PCR assay for authenticity control in farm animal feeds and pet foods.

    Science.gov (United States)

    Pegels, Nicolette; González, Isabel; García, Teresa; Martín, Rosario

    2014-01-01

    A highly sensitive TaqMan real-time PCR assay targeting the mitochondrial 12S rRNA gene was developed for detection of an avian-specific DNA fragment (68bp) in farm animal and pet feeds. The specificity of the assay was verified against a wide representation of animal and plant species. Applicability assessment of the avian real-time PCR was conducted through representative analysis of two types of compound feeds: industrial farm animal feeds (n=60) subjected to extreme temperatures, and commercial dog and cat feeds (n=210). Results obtained demonstrated the suitability of the real-time PCR assay to detect the presence of low percentages of highly processed avian material in the feed samples analysed. Although quantification results were well reproducible under the experimental conditions tested, an accurate estimation of the target content in feeds is impossible in practice. Nevertheless, the method may be useful as an alternative tool for traceability purposes within the framework of feed control.

  8. Subtyping Animal Influenza Virus with General Multiplex RT-PCR and Liquichip High Throughput (GMPLex)

    Institute of Scientific and Technical Information of China (English)

    Zhi-feng Qin; Bing Cheng; Zhou-xi Ruan; Ying-zuo Bi; Joseph J Giambrone; Hong-zhuan Wu; Jie Sun; Ti-kang Lu; Shao-ling Zeng; Qun-yi Hua; Qing-yan Ling; Shu-kun Chen; Jian-qiang Lv; Cai-hong Zhang

    2012-01-01

    This study developed a multiplex RT-PCR integrated with luminex technology to rapidly subtype simultaneously multiple influenza viruses.Primers and probes were designed to amplify NS and M genes of influenza A viruses HA gene of H1,H3,H5,H7,H9 subtypes,and NA gene of the N1 and N2 subtypes.Universal super primers were introduced to establish a multiplex RT-PCR (GM RT-PCR).It included three stages of RT-PCR amplification,and then the RT-PCR products were further tested by LiquiChip probe,combined to give an influenza virus (Ⅳ) rapid high throughput subtyping test,designated as GMPLex.The IV GMPLex rapid high throughput subtyping test presents the following features:high throughput,able to determine the subtypes of 9 target genes in H1,H3,H5,H7,H9,N1,and N2 subtypes of the influenza A virus at one time; rapid,completing the influenza subtyping within 6 hours; high specificity,ensured the specificity of the different subtypes by using two nested degenerate primers and one probe,no cross reaction occurring between the subtypes,no non-specific reactions with other pathogens and high sensitivity.When used separately to detect the product of single GM RT-PCR for single H5 or N1 gene,the GMPLex test showed a sensitivity of 105(=280ELD50) forboth tests and the Luminex qualitative ratio results were 3.08 and 3.12,respectively.When used to detect the product of GM RT-PCR for HSN1 strain at the same time,both showed a sensitivity of 10-4(=2800 ELD50).The GMPLex rapid high throughput subtyping test can satisfy the needs of influenza rapid testing.

  9. Comparison of electron microscopy, ELISA, real time RT-PCR and insulated isothermal RT-PCR for the detection of Rotavirus group A (RVA) in feces of different animal species.

    Science.gov (United States)

    Soltan, Mohamed A; Tsai, Yun-Long; Lee, Pei-Yu A; Tsai, Chuan-Fu; Chang, Hsiao-Fen G; Wang, Hwa-Tang T; Wilkes, Rebecca P

    2016-09-01

    There is no gold standard for detection of Rotavirus Group A (RVA), one of the main causes of diarrhea in neonatal animals. Sensitive and specific real-time RT-PCR (rtRT-PCR) assays are available for RVA but require submission of the clinical samples to diagnostic laboratories. Patient-side immunoassays for RVA protein detection have shown variable results, particularly with samples from unintended species. A sensitive and specific test for detection of RVA on the farm would facilitate rapid management decisions. The insulated isothermal RT-PCR (RT-iiPCR) assay works in a portable machine to allow sensitive and specific on-site testing. The aim of this investigation was to evaluate a commercially available RT-iiPCR assay for RVA detection in feces from different animal species. This assay was compared to an in-house rtRT-PCR assay and a commercially available rtRT-PCR kit, as well as an ELISA and EM for RVA detection. All three PCR assays targeted the well-conserved NSP5 gene. Clinical fecal samples from 108 diarrheic animals (mainly cattle and horses) were tested. The percentage of positive samples by ELISA, EM, in-house rtRT-PCR, commercial rtRT-PCR, and RT-iiPCR was 29.4%, 31%, 36.7%, 51.4%, 56.9%, respectively. The agreement between different assays was high (81.3-100%) in samples containing high viral loads. The sensitivity of the RT-iiPCR assay appeared to be higher than the commercially available rtRT-PCR assay, with a limit of detection (95% confidence index) of 3-4 copies of in vitro transcribed dsRNA. In conclusion, the user-friendly, field-deployable RT-iiPCR system holds substantial promise for on-site detection of RVA.

  10. Forensic animal DNA analysis using economical two-step direct PCR.

    Science.gov (United States)

    Kitpipit, Thitika; Chotigeat, Wilaiwan; Linacre, Adrian; Thanakiatkrai, Phuvadol

    2014-03-01

    Wildlife forensic DNA analysis by amplification of a mitochondrial locus followed by DNA sequencing is routine, yet suffers from being costly and time-consuming. To address these disadvantages we report on a low-cost two-step direct PCR assay to efficiently analyze 12 forensically relevant mammalian sample types without DNA extraction. A cytochrome oxidase I degenerate-universal primer pair was designed and validated for the developed assay. The 12 sample types, which included bone, horn, feces, and urine, were amplified successfully by the assay using a pre-direct PCR dilution protocol. The average amplification success rate was as high as 92.5 % (n = 350), with an average PCR product concentration of 220.71 ± 180.84 ng/μL. Differences in amplification success rate and PCR product quantity between sample types were observed; however, most samples provided high quality sequences, permitting a 100 % nucleotide similarity to their respective species via BLAST database queries. The combination of PBS and Phire(®) Hot Start II DNA polymerase gave comparable amplification success rate and amplicon quantity with the proprietary commercial kits (P > 0.05, n = 350) but at considerably lower cost. The stability of the assay was tested by successfully amplifying samples that had been stored for up to 12 months. Our data indicate that this low-cost two-step direct amplification assay has the potential to be a valuable tool for the forensic DNA community.

  11. Protocolo de extracción de ADN en lotes de 10 mosquitos para la identificación de Plasmodium spp. mediante qPCR

    Directory of Open Access Journals (Sweden)

    A. Pérez Rico

    Full Text Available Las tropas que despliegan en zonas de operaciones endémicas de malaria, necesitan de una información precisa del riesgo sanitario para la toma de decisiones acerca de las medidas de prevención más adecuadas. El estado de portador de un mosquito se determina clásicamente por la presencia o ausencia de esporozoitos de Plasmodium spp. en las glándulas salivales. Los protocolos basados en la amplificación del ADN en tiempo real (qPCR son muy sensibles, sin embargo existen dificultades en la qPCR debido a inhibidores presentes en los tejidos del mosquito, lo que obliga a trabajar de uno en uno. En este trabajo se diseña una qPCR para amplificar una región conservada entre mosquitos de diferentes especies y otros dípteros, con el objetivo de comparar varios protocolos de extracción de ADN y determinar el más eficiente a la hora de procesar lotes de 10 mosquitos.

  12. Comparison of bacterial culture and polymerase chain reaction (PCR) for the detection of F. tularensis subsp. holarctica in wild animals.

    Science.gov (United States)

    Sting, Reinhard; Runge, Martin; Eisenberg, Tobias; Braune, Silke; Müller, Wolfgang; Otto, Peter

    2013-01-01

    Detection of the zoonotic pathogen Francisella tularensis subsp. holarctica (EF tularensis) in wild animals with culture techniques as well as polymerase chain reaction were compared and discussed on the basis of the investigation of 60 animals. The samples originated from 55 European brown hares (Lepus europaeus), two red foxes (Vulpes vulpes) and one each from a wild rabbit (Oryctolagus cuniculus), a European beaver (Castor fiber), and a lemur (Lemur catta). When comparing the growth of 28 F. tularensis isolates on the cysteine blood agar and the modified Martin-Lewis-agar used in this study, cultivation was successful for 26 isolates on both media, but for two isolates only on the cysteine blood agar. Out of 43 carcasses 19 tested positive in bacteriological culture and PCR. Two culture positive samples of tonsils originating from foxes could not be confirmed by PCR, although PCR was positive in 22 samples that missed growth of F. tularensis. Comparative studies on cultural detection of E. tularensis were performed on samples of 16 hares from lung, spleen, liver and gut and in one case with a peritoneal swab. In at least one of these localizations cultivation of the pathogen was successful. Detection rate was reduced to 94% (15 of 16 hares) considering only the results of the cultures of the lungs and spleens. For a sensitive and rapid detection of F. tularensis subsp. holarctica, the PCR is a suitable method thereby avoiding hazardous multiplying of the pathogen. However, cultivation of F. tularensis is often a prerequisite for further studies on antibiotic resistance patterns of the pathogen, molecular epidemiological and pathological analyses of tularaemia.

  13. Quantificazione mediante PCR dell’EBV-DNA da biopsie cutanee di pazienti con linfomi cutanei primitivi (micosi fungoide e sindrome di Sèzary

    Directory of Open Access Journals (Sweden)

    Chiara Merlino

    2007-06-01

    Full Text Available Mycosis fungoides (MF, the most indolent form of CTCL, originates from a clonal expansion of epidermotropic helper/memory T cells. Sezary syndrome (SS is a rare primay epidermotropic cutaneous T-cell lymphoma in leukemic. The aetiopathogenesis of MF and SS remains obscure despite several investigations. Infectious, environmental and genetic factors have been implicated as potential aetiological agents. The studies investigating the role of EBV in CTCL present conflicting results. The different sensitivities of the technical methods used in the evaluation of the presence of viral DNA or virus-related antigens make comparison of the results difficult. The aim of this study was to retrospectively evaluate the EBV-DNA load in skin biopsies from MF and SS patients by a highly sensitive (1-10 EBV-DNA copies/reaction quantitative-competitive PCR (QC-PCR developed in our lab to better asses the relationship between EBV and CTCL. Skin biopsies were obtained from 21 MF and 10 SS patients; skin biopsies from a 8 patients with inflammatory skin disease were used as controls. EBV-DNA was detected in 70% of biopsies from SS patients vs. 0% of MF patients. No control patients resulted EBV-DNA positive, as expected. In addition, in SS patients, the survival from diagnosis is lesser in EBV-positive patients vs.EBV-negative patients even if not statistically significant.We are going to investigate the presence of EBV-DNA in peripheral blood of a larger number of patients and to evaluate the pattern of viral genes expression, to better assess the aetiopathogenetical role of EB virus in this kind of neoplasies.

  14. Enhanced detection of tuberculous mycobacteria in animal tissues using a semi-nested probe-based real-time PCR.

    Directory of Open Access Journals (Sweden)

    Pedro Costa

    Full Text Available Bovine tuberculosis has been tackled for decades by costly eradication programs in most developed countries, involving the laboratory testing of tissue samples from allegedly infected animals for detection of Mycobacterium tuberculosis complex (MTC members, namely Mycobacterium bovis. Definitive diagnosis is usually achieved by bacteriological culture, which may take up to 6-12 weeks, during which the suspect animal carcass and herd are under sanitary arrest. In this work, a user-friendly DNA extraction protocol adapted for tissues was coupled with an IS6110-targeted semi-nested duplex real-time PCR assay to enhance the direct detection of MTC bacteria in animal specimens, reducing the time to achieve a diagnosis and, thus, potentially limiting the herd restriction period. The duplex use of a novel β-actin gene targeted probe, with complementary targets in most mammals, allowed the assessment of amplification inhibitors in the tissue samples. The assay was evaluated with a group of 128 fresh tissue specimens collected from bovines, wild boars, deer and foxes. Mycobacterium bovis was cultured from 57 of these samples. Overall, the full test performance corresponds to a diagnostic sensitivity and specificity of 98.2% (CIP95% 89.4-99.9% and 88.7% (CIP95% 78.5-94.7%, respectively. An observed kappa coefficient was estimated in 0.859 (CI P95% 0.771-0.948 for the overall agreement between the semi-nested PCR assay and the bacteriological culture. Considering only bovine samples (n = 69, the diagnostic sensitivity and specificity were estimated in 100% (CIP95% 84.0-100% and 97.7% (CIP95% 86.2-99.9%, respectively. Eight negative culture samples exhibiting TB-like lesions were detected by the semi-nested real-time PCR, thus emphasizing the increased potential of this molecular approach to detect MTC-infected animal tissues. This novel IS6110-targeted assay allows the fast detection of tuberculous mycobacteria in animal specimens with very high

  15. Enhanced detection of tuberculous mycobacteria in animal tissues using a semi-nested probe-based real-time PCR.

    Science.gov (United States)

    Costa, Pedro; Ferreira, Ana S; Amaro, Ana; Albuquerque, Teresa; Botelho, Ana; Couto, Isabel; Cunha, Mónica V; Viveiros, Miguel; Inácio, João

    2013-01-01

    Bovine tuberculosis has been tackled for decades by costly eradication programs in most developed countries, involving the laboratory testing of tissue samples from allegedly infected animals for detection of Mycobacterium tuberculosis complex (MTC) members, namely Mycobacterium bovis. Definitive diagnosis is usually achieved by bacteriological culture, which may take up to 6-12 weeks, during which the suspect animal carcass and herd are under sanitary arrest. In this work, a user-friendly DNA extraction protocol adapted for tissues was coupled with an IS6110-targeted semi-nested duplex real-time PCR assay to enhance the direct detection of MTC bacteria in animal specimens, reducing the time to achieve a diagnosis and, thus, potentially limiting the herd restriction period. The duplex use of a novel β-actin gene targeted probe, with complementary targets in most mammals, allowed the assessment of amplification inhibitors in the tissue samples. The assay was evaluated with a group of 128 fresh tissue specimens collected from bovines, wild boars, deer and foxes. Mycobacterium bovis was cultured from 57 of these samples. Overall, the full test performance corresponds to a diagnostic sensitivity and specificity of 98.2% (CIP95% 89.4-99.9%) and 88.7% (CIP95% 78.5-94.7%), respectively. An observed kappa coefficient was estimated in 0.859 (CI P95% 0.771-0.948) for the overall agreement between the semi-nested PCR assay and the bacteriological culture. Considering only bovine samples (n = 69), the diagnostic sensitivity and specificity were estimated in 100% (CIP95% 84.0-100%) and 97.7% (CIP95% 86.2-99.9%), respectively. Eight negative culture samples exhibiting TB-like lesions were detected by the semi-nested real-time PCR, thus emphasizing the increased potential of this molecular approach to detect MTC-infected animal tissues. This novel IS6110-targeted assay allows the fast detection of tuberculous mycobacteria in animal specimens with very high sensitivity and

  16. Animal model for training in sentinel lymph node biopsy of the stomach through combined methods Modelo animal para treinamento em pesquisa de linfonodo sentinela em estômago mediante métodos combinados

    Directory of Open Access Journals (Sweden)

    José Roberto Alves

    2012-12-01

    Full Text Available PURPOSE: Create and validate a proposed animal model for training in sentinel lymph node biopsy of the stomach. METHODS: In thirty-two rabbits, through a laparotomy, they received a subserosal injection of 0.1 ml of phytate labeled with technetium-99m (0.2 mCi in the anterior wall of the gastric corpus, followed by 0.2 ml of Blue Patent V® 2.5%, through the same puncture site. Suspicious lymph nodes were searched in vivo at five, ten and 20 minutes, both visually (Blue Patent stained lymph nodes and with a manual gamma radiation detector (to detect suspected radioactive lymph nodes. After 20 minutes, was performed resection of these for further evaluation of radioactivity (ex vivo and histological study. RESULTS: Lymph nodes were identified in 30 rabbits (Average of 2.2 lymph nodes per animal. Of the 90 suspected lymph nodes that occurred in the study, 70 cases (77.8% were histologically confirmed for lymphoid tissue. Of these, the majority were located in the periesophageal region of the gastric fundus. The sample presented a mortality rate of 6.25% and nine complications related to the method, which interfered in the identification of the lymph nodes. CONCLUSION: The animal model for sentinel node biopsy in rabbit stomachs proved to be feasible, with low complexity and reproduced the difficulties encountered for gastric lymph node biopsy in humans, being adequate for surgical training.OBJETIVO: Criar e validar uma proposta de modelo animal para o treinamento em pesquisa de linfonodos sentinelas no estômago. MÉTODOS: Em trinta e dois coelhos, mediante laparotomia, foi injetado na subserosa da parede anterior do corpo gástrico, 0,1 ml de fitato marcado com tecnécio-99m (0,2 mCi, seguido pelo mesmo orifício, de 0,2 ml de Azul Patente V® 2,5%. A cavidade abdominal foi avaliada, in vivo, por meio de inspeção para pesquisa de suspeitas de linfonodos azuis e com detector manual de radiação gamma aos cinco, dez e 20 minutos para pesquisa de

  17. Validation of Bacteroidales quantitative PCR assays targeting human and animal fecal contamination in the public and domestic domains in India.

    Science.gov (United States)

    Odagiri, Mitsunori; Schriewer, Alexander; Hanley, Kaitlyn; Wuertz, Stefan; Misra, Pravas R; Panigrahi, Pinaki; Jenkins, Marion W

    2015-01-01

    We compared host-associated Bacteroidales qPCR assays developed in the continental United States and Europe for the purpose of measuring the effect of improved sanitation on human fecal exposure in rural Indian communities where both human and animal fecal loading are high. Ten candidate Bacteroidales qPCR assays were tested against fecal samples (human, sewage, cow, buffalo, goat, sheep, dog and chicken) from a test set of 30 individual human, 5 sewage, and 60 pooled animal samples collected in coastal Odisha, India. The two universal/general Bacteroidales assays tested (BacUni, GenBac3) performed equally well, achieving 100% sensitivity on the test set. Across the five human-associated assays tested (HF183 Taqman, BacHum, HumM2, BacH, HF183 SYBR), we found low sensitivity (17 to 49%) except for HF183 SYBR (89%), and moderate to high cross-reactivity with dog (20 to 80%) and chicken fecal samples (60 to 100%). BacHum had the highest accuracy (67%), amplified all sewage samples within the range of quantification (ROQ), and did not cross-react with any fecal samples from cows, the most populous livestock animal in India. Of the ruminant- and cattle-associated assays tested (BacCow, CowM2), BacCow was more sensitive in detecting the full range of common Indian livestock animal fecal sources, while CowM2 only detected cow sources with 50% sensitivity. Neither assay cross-reacted with human sources. BacCan, the dog-associated assay tested, showed no cross-reactivity with human sources, and high sensitivity (90%) for dog fecal samples. Overall, our results indicate BacUni, BacHum, HumM2, BacCan and BacCow would be the most suitable MST assays to distinguish and quantify relative amounts of human-associated and livestock/domestic animal-associated contributions to fecal contamination in Odisha, India.

  18. Evaluation of pre-PCR processing approaches for enumeration of Salmonella enterica in naturally contaminated animal feed

    DEFF Research Database (Denmark)

    Schelin, Jenny; Andersson, Gunnar; Vigre, Håkan

    2014-01-01

    Three pre‐PCR processing strategies for the detection and/or quantification of Salmonella in naturally contaminated soya bean meal were evaluated. Methods included: (i) flotation‐qPCR [enumeration of intact Salmonella cells prior to quantitative PCR (qPCR)], (ii) MPN‐PCR (modified most probable...... number method combined with qPCR) and (iii) qualitative culture enrichment PCR. The limit of quantification was 1·8 × 102 CFU g−1 (flotation‐qPCR) and 0·02 MPN g−1 (MPN‐PCR). Fifteen naturally contaminated Salmonella positive soya bean meal samples from one lot were analysed in parallel with the three...

  19. Giardia duodenalis genotypes in domestic and wild animals from Romania identified by PCR-RFLP targeting the gdh gene.

    Science.gov (United States)

    Adriana, Gyӧrke; Zsuzsa, Kalmár; Mirabela Oana, Dumitrache; Mircea, Gherman Călin; Viorica, Mircean

    2016-02-15

    Sixty Giardia duodenalis isolates from domestic (n=49) and wild (n=11) animals (dogs, cats, deers, wolves, raccoon dog and muskrat) were analysed by PCR-RFLP at glutamate dehydrogenase locus (gdh). The isolates were obtained from positive feces samples for Giardia cysts analysed by flotation technique with saturated sodium chloride solution (specific gravity 1.28). Three G. duodenalis genotypes were identified: C (10/60; 16.7%); D (42/60; 70.0%); and E (7/60; 11.7%). In dogs all three genotypes were found, with the following prevalences: 76.9% genotype D (30/39); 23.1% C (9/39); 2.6% genotype E (1/39). One dog was co-infected with C and D genotypes. In cats we identified only G. duodenalis genotype D. Wolves and raccoon dog harbored infection with G. duodenalis genotype D, deers with E type and muskrat C type. This is the first study regarding genotyping of G. duodenalis in cats and wild animals from Romania. To the best of our knowledge, this is the first report of assemblages E in roe deers; assemblage C in wolves and muskrat; and assemblage D in raccoon dog.

  20. Análisis de los componentes de la respuesta a la selección en un núcleo con generaciones superpuestas seleccionado mediante BLUP bajo un modelo animal

    OpenAIRE

    Schindler de Avila, Valeria

    1996-01-01

    p.39-48 Con el objetivo de analizar la importancia relativa de los componentes de la respuesta a la selección, se simuló un núcleo con generaciones superpuestas durante 20 años. Se estudió un carácter medible en ambos sexos y una única vez durante la vida del animal. Se estudiaron d o sh² del carácter: 0,4 y 0,1. Los animales se seleccionaron mediante BLUP-modelo animal, incluyendo los efectos fijos del sexo y la edad del individuo a la medición. La respuesta a la selección fue siempre may...

  1. Presence of a Phytoplasma Associated with Witches’-Broom Disease in Ugni molinae Turcz. and Gaultheria phillyreifolia (Pers. Sleumer Determined by DAPI, PCR, and DNA Sequencing Presencia de un Fitoplasma Asociado a la Enfermedad de "Escoba de Bruja" en Ugni molinae Turcz. y Gaultheria phillyreifolia (Pers. Sleumer Determinado Mediante DAPI, PCR y Secuenciación de ADN

    Directory of Open Access Journals (Sweden)

    Nolberto Arismendi S

    2010-03-01

    Full Text Available Murta (Ugni molinae Turcz. and common chaura (Gaultheria phillyreifolia (Pers. Sleumer are native species of Chile. Plants of both species have shown over-branching like witches' broom. The causal agents of these symptoms in many plants are phytoplasma. To verify the presence of these microorganisms, DAPI (4',6-diamidino-2-phenylindole staining analysis and polymerase chain reaction (PCR were performed in symptomatic and asymptomatic plants. Positive PCR samples were sequenced to identify the pathogens involved. In individuals of both species with witches’ broom symptoms, DAPI staining showed fluorescent bodies in the phloem tissues, but not in asymptomatic plants. Verification by nested-PCR, phytoplasmatic DNA was amplified from diseased murta and chaura, but not in apparently healthy plants. Sequencing of amplified products allowed locating phytoplasma within the ash yellows group (16SrVII and related to Candidatus phytoplasma fraxini. This is the first report of phytoplasma in Chilean native species. Considering the diversity of plant species infected by the ash yellows group suggests that G. phillyreifolia and U. molinae could be a phytoplasma reservoir for other economically important agricultural crops.La murta (Ugni molinae Turcz. y la chaura común (Gaultheria phillyreifolia (Pers. Sleumer son especies nativas de Chile. En plantas de ambas especies se ha observado una sobre-ramificación de tipo "escoba de bruja". En muchas plantas los agentes causales de esta sintomatología son fitoplasmas. Para verificar la presencia de estos microorganismos se analizaron plantas con y sin síntomas mediante tinciones DAPI (4’,6-diamidino-2-fenilindol y reacción en cadena de la polimerasa (PCR. Muestras positivas en la PCR fueron secuenciadas para identificar al fitopatógeno implicado. En individuos de ambas especies con síntomas de escoba de bruja, la tinción DAPI permitió observar cuerpos fluorescentes en los tejidos del floema, situaci

  2. Development and validation of a real-time PCR assay for the detection of Toxoplasma gondii DNA in animal and meat samples.

    Science.gov (United States)

    Marino, Anna Maria Fausta; Percipalle, Maurizio; Giunta, Renato Paolo; Salvaggio, Antonio; Caracappa, Giulia; Alfonzetti, Tiziana; Aparo, Alessandra; Reale, Stefano

    2017-03-01

    We report a rapid and reliable method for the detection of Toxoplasma gondii in meat and animal tissues based on real-time polymerase chain reaction (PCR). Samples were collected from cattle, small ruminants, horses, and pigs raised or imported into Sicily, Italy. All DNA preparations were assayed by real-time PCR tests targeted to a 98-bp long fragment in the AF 529-bp repeat element and to the B1 gene using specific primers. Diagnostic sensitivity (100%), diagnostic specificity (100%), limit of detection (0.01 pg), efficiency (92-109%), and precision (mean coefficient of variation = 0.60%), repeatability (100%), reproducibility (100%), and robustness were evaluated using 240 DNA extracted samples (120 positives and 120 negative as per the OIE nested PCR method) from different matrices. Positive results were confirmed by the repetition of both real-time and nested PCR assays. Our study demonstrates the viability of a reliable, rapid, and specific real-time PCR on a large scale to monitor contamination with Toxoplasma cysts in meat and animal specimens. This validated method can be used for postmortem detection in domestic and wild animals and for food safety purposes.

  3. Identification of new flagellin-encoding fliC genes in Escherichia coli isolated from domestic animals using RFLP-PCR and sequencing methods

    Directory of Open Access Journals (Sweden)

    Cláudia de Moura

    2013-04-01

    Full Text Available Identification of Escherichia coli requires knowledge regarding the prevalent serotypes and virulence factors profiles allows the classification in pathogenic/non-pathogenic. However, some of these bacteria do not express flagellar antigen invitro. In this case the PCR-restriction fragment length polymorphism (RFLP-PCR and sequencing of the fliC may be suitable for the identification of antigens by replacing the traditional serology. We studied 17 samples of E. coli isolated from animals and presenting antigen H nontypeable (HNT. The H antigens were characterized by PCR-RFLP and sequencing of fliC gene. Three new flagellin genes were identified, for which specific antisera were obtained. The PCR-RFLP was shown to be faster than the serotyping H antigen in E. coli, provided information on some characteristics of these antigens and indicated the presence of new genes fliC.

  4. Animals

    Energy Technology Data Exchange (ETDEWEB)

    Skuterud, L.; Strand, P. [Norwegian Radiation Protection Authority (Norway); Howard, B.J. [Inst. of Terrestrial Ecology (United Kingdom)

    1997-10-01

    The radionuclides of most concern with respect to contamination of animals after a nuclear accident are radioiodine, radiocaesium and radiostrontium (ICRP 30, 1979). Of the other significant anthropogenic radionuclides likely to be released in most accidents, only small proportions of that ingested will be absorbed in an animals gut, and the main animal products, milk and meat, will not normally be contaminated to a significant extent. Animal products will mostly be contaminated as a result of ingestion of contaminated feed and possibly, but to a much lesser extent, from inhalation (for radioiodine only). Direct external contamination of animals is of little or no consequence in human food production. Radioiodine and radiostrontium are important with respect to contamination of milk; radiocaesium contaminates both milk and meat. The physical and chemical form of a radionuclide can influence its absorption in the animal gut. For example, following the Chernobyl accident radiocaesium incorporated into vegetation by root uptake was more readily absorbed than that associated with the original deposit. The transfer of radiocaesium and radiostrontium to animals will be presented both as transfer coefficients and aggregated transfer coefficients. For most animal meat products, only radiocaesium is important as other radionuclides do not significantly contaminate muscle. Farm animal products are the most important foodstuff determining radiocaesium intake by the average consumer in the Nordic countries. The major potential source of radioiodine and radiostrontium to humans is milk and milk products. Of the different species, the smaller animals have the highest transfer of radiocaesium from fodder to meat and milk. (EG). 68 refs.

  5. Automated Forensic Animal Family Identification by Nested PCR and Melt Curve Analysis on an Off-the-Shelf Thermocycler Augmented with a Centrifugal Microfluidic Disk Segment.

    Science.gov (United States)

    Keller, Mark; Naue, Jana; Zengerle, Roland; von Stetten, Felix; Schmidt, Ulrike

    2015-01-01

    Nested PCR remains a labor-intensive and error-prone biomolecular analysis. Laboratory workflow automation by precise control of minute liquid volumes in centrifugal microfluidic Lab-on-a-Chip systems holds great potential for such applications. However, the majority of these systems require costly custom-made processing devices. Our idea is to augment a standard laboratory device, here a centrifugal real-time PCR thermocycler, with inbuilt liquid handling capabilities for automation. We have developed a microfluidic disk segment enabling an automated nested real-time PCR assay for identification of common European animal groups adapted to forensic standards. For the first time we utilize a novel combination of fluidic elements, including pre-storage of reagents, to automate the assay at constant rotational frequency of an off-the-shelf thermocycler. It provides a universal duplex pre-amplification of short fragments of the mitochondrial 12S rRNA and cytochrome b genes, animal-group-specific main-amplifications, and melting curve analysis for differentiation. The system was characterized with respect to assay sensitivity, specificity, risk of cross-contamination, and detection of minor components in mixtures. 92.2% of the performed tests were recognized as fluidically failure-free sample handling and used for evaluation. Altogether, augmentation of the standard real-time thermocycler with a self-contained centrifugal microfluidic disk segment resulted in an accelerated and automated analysis reducing hands-on time, and circumventing the risk of contamination associated with regular nested PCR protocols.

  6. Evaluación madurativo-mental y emocional en sujetos repetidores mediante el test del dibujo animal y el autoconcepto

    Directory of Open Access Journals (Sweden)

    Juana María MAGANTO MATEO

    2009-11-01

    Full Text Available El presente trabajo se enmarca en un contexto psicopedagógico que integra dos componentes fundamentales: los niños que fracasan escolarmente y la evaluación descriptiva- comprensiva de los mismos. Las bases teórico-prácticas sobre las que se asienta este estudio están por un lado conectadas con las investigaciones sobre el fracaso escolar, y por otro con las técnicas diagnósticas gráficas susceptibles de evaluación sistemática y comparativa. En concreto, el tema central se focalÍ2a en el diagnóstico desde el punto de vista evolutivo, a través de un test gráfico denominado Test del Dibujo del Animal, de los sujetos en los que su fracaso escolar ha derivado en la repetición del curso. Se ha desarrollado, y actualmente prosiguen, numerosos estudios sobre el fracaso escolar. El marco teórico desde el cual se parte aporta perspectivas diversas de comprensión y análisis del mismo.

  7. Animals

    Institute of Scientific and Technical Information of China (English)

    杨光

    2000-01-01

    The largest animal ever to live on the earth is the blue whale(蓝鲸)It weighs about 80 tons--more than 24 elephants. It is more than 30 metres long. A newborn baby whale weighs as much as a big elephant.

  8. Shedding of Clostridium difficile PCR ribotype 078 by zoo animals, and report of an unstable metronidazole-resistant isolate from a zebra foal (Equus quagga burchellii).

    Science.gov (United States)

    Álvarez-Pérez, Sergio; Blanco, José L; Martínez-Nevado, Eva; Peláez, Teresa; Harmanus, Celine; Kuijper, Ed; García, Marta E

    2014-03-14

    Clostridium difficile is an emerging and potentially zoonotic pathogen, but its prevalence in most animal species, including exhibition animals, is currently unknown. In this study we assessed the prevalence of faecal shedding of C. difficile by zoo animals, and determined the ribotype, toxin profile and antimicrobial susceptibility of recovered isolates. A total of 200 samples from 40 animal species (36.5% of which came from plains zebra, Equus quagga burchellii) were analysed. C. difficile was isolated from 7 samples (3.5% of total), which came from the following animal species: chimpanzee (Pan troglodytes troglodytes), dwarf goat (Capra hircus), and Iberian ibex (Capra pyrenaica hispanica), with one positive sample each; and plains zebra, with 4 positive samples from 3 different individuals. Most recovered isolates (4/7, 57.1%) belonged to the epidemic PCR ribotype 078, produced toxins A and B, and had the genes encoding binary toxin (i.e. A(+)B(+)CDT(+) isolates). The remaining three isolates belonged to PCR ribotypes 039 (A(-)B(-)CDT(-)), 042 (A(+)B(+)CDT(-)) and 110 (A(-)B(+)CDT(-)). Regardless of their ribotype, all isolates displayed high-level resistance to the fluoroquinolones ciprofloxacin, enrofloxacin and levofloxacin. Some isolates were also resistant to meropenem and/or ertapenem. A ribotype 078 isolate recovered from a male zebra foal initially showed in vitro resistance to metronidazole (MIC ≥ 256 μg/ml), but lost that trait after subculturing on non-selective media. We conclude that zoo animals belonging to different species can carry ribotype 078 and other toxigenic strains of C. difficile showing resistance to antimicrobial compounds commonly used in veterinary and/or human medicine.

  9. Comparison of PCR-Based Diagnosis with Centrifuged-Based Enrichment Method for Detection of Borrelia persica in Animal Blood Samples

    Directory of Open Access Journals (Sweden)

    SR Naddaf

    2011-06-01

    Background: The mainstay of diagnosis of relapsing fever (RF is demonstration of the spirochetes in Giemsa-stained thick blood smears, but during non fever periods the bacteria are very scanty and rarely detected in blood smears by mi­cros­copy. This study is aimed to evaluate the sensitivity of different methods developed for detection of low-grade spi­ro­chetemia. Methods: Animal blood samples with low degrees of spirochetemia were tested with two PCRs and a nested PCR tar­get­ing flaB, GlpQ, and rrs genes. Also, a centrifuged-based enrichment method and Giemsa staining were per­formed on blood samples with various degrees of spirochetemia. Results: The flaB-PCR and nested rrs-PCR turned positive with various degrees of spirochetemia including the blood samples that turned negative with dark-field microscopy. The GlpQ-PCR was positive as far as at least one spi­ro­chete was seen in 5-10 microscopic fields. The sensitivity of GlpQ-PCR increased when DNA from Buffy Coat Layer (BCL was used as template. The centrifuged-based enrichment method turned positive with as low concentra­tion as 50 bacteria/ml blood, while Giemsa thick staining detected bacteria with concentrations ≥ 25000 bacteria/ml.  Conclusion: Centrifuged-based enrichment method appeared as much as 500-fold more sensitive than thick smears, which makes it even superior to some PCR assays. Due to simplicity and minimal laboratory requirements, this method can be considered a valuable tool for diagnosis of RF in rural health centers.  

  10. Comparison of DNA isolation methods and detection of Salmonella spp. from animal faeces and dust using invA real-time PCR.

    Science.gov (United States)

    Braun, Sascha D; Methner, Ulrich

    2011-01-01

    There is a strong interest to reduce the expenditure for the detection of Salmonella spp. from animal faeces and environmental samples from primary production according to ISO 6579:2002 Annex D by including a rapid and effective method to detect Salmonella spp. already after pre-enrichment in BPW. It has been shown that real-time PCR methods are very effective to detect Salmonella organisms after pre-enrichment of foods. However, materials from primary animal production compose of much higher amounts of substances which might inhibit the sensitivity of real-time PCR. Different techniques of DNA isolation after pre-enrichment of artificially inoculated bovine faecal material were used to compare their detection limit and detection probability using an invA 5' nuclease real-time PCR approach. A detection probability of 100% was shown at 10(5) cfu/ml using the QIAamp DNA Stool Mini Kit (Qiagen, Germany), at 10(4) cfu/ml using the High Pure PCR Template Preparation Kit (Roche, Germany) and at 10(3) cfu/ml using thermal cell lysis or an in-house lab protocol, respectively. In comparison DNA isolation by thermal cell lysis revealed a very good detection limit, low costs and almost no risks of contamination. Furthermore, caecal contents from pigs were analysed by ISO 6579:2002 Annex D and the invA real-time PCR using thermal cell lysis for DNA extraction. As a result neither false positive nor false negative findings were obtained. Inclusion of the real-time PCR after pre-enrichment of samples in BPW followed by bacterial detection of Salmonella only with samples positive with real-time PCR might be a valuable tool to fulfil the international standard of ISO 6579:2002 Annex D but also to diminish the expenditures. However, it must be stated that the modification of an international standard method and its use in routine diagnostic requires the validation and registration of national and/or international competent authorities.

  11. Validation of an open-formula, diagnostic real-time PCR method for 20-hr detection of Salmonella in animal feeds

    DEFF Research Database (Denmark)

    Löfström, Charlotta; Hoorfar, Jeffrey

    2012-01-01

    A comparative study of a 20-hr, non-commercial, open-formula PCR method and the standard culture-based method NMKL 187, for detection of Salmonella, was performed according to the validation protocol from the Nordic organization for validation of alternative microbiological methods (NordVal) on 81...... artificially or naturally contaminated animal feed samples. The PCR method is based on culture enrichment in buffered peptone water for 16 ± 2 h followed by a magnetic beads based semi automated DNA extraction and real-time PCR analysis, including an internal amplification control. The limit of detection (LOD......50) was found to be 7.19 and 7.24 CFU/sample for the PCR method and NMKL187, respectively. A very good correlation between results obtained by the two methods were found (Coheńs kappa = 0.92). The relative accuracy, relative sensitivity and relative specificity were found to be 97.5%, 102.0% and 96...

  12. Identification of Echinococcus Granulosus Strains in Isolated Hydatid Cyst Specimens from Animals by PCR-RFLP Method in West Azerbaijan – Iran

    Directory of Open Access Journals (Sweden)

    Haleh Hanifian

    2013-09-01

    Full Text Available Background: The aim of this study was DNA extraction from protosco­lecses of Echinococcus granulosus and identification of these strains in West-Azerbai­jan Province, north western Iran.Methods: Thirty one livestock isolates from sheep and cattle were collected from abattoirs of the province. To investigate the genetic variation of the isolates, after DNA extraction by Glass beads-phenol chloroform method; PCR-RLFP analysis of rDNA-ITS1 was performed using three different restric­tion enzymes of Taq 1, Rsa 1 and Alu 1.Result: Amplified PCR products for all isolates were 1000bp band which is expected band in sheep strains (G1-G3 complex. The results of RFLP analy­sis also were the same for all isolates. PCR-RFLP patterns restriction en­zymes were identical as follows, Rsa1 bands under UV showed two bands approximately 655bp and 345bp. Alu1 bands were as follows: two approx­imately 800bp and 200bp and Taq1 did not cut any region and bands were approximately 1000 bp in all samples.Conclusions: Based on PCR-RFLP patterns of ITS1 fragment produced with endonucleases enzyme digestion in animal isolates, it can be concluded that a single strain of E. granulosus (sheep strain or G1-G3 complex is domi­nant genotype in this province

  13. ANIMALS

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Mammals(哺乳动物)Mammals are the world's most dominant(最占优势的)animal.They are extremely(非常)diverse(多种多样的)creatures(生物,动物)that include(包括)the biggest ever animal (the blue whale鲸,which eats up to 6 tons every day),the smallest(leaf-nosed bat小蹄蝠) and the laziest(sloth树獭,who spends 80% of their time sleeping).There are over 4,600 kinds of mammals and they live in very different environments(环境)—oceans(海洋),rivers,the jungle(丛林),deserts,and plains(平原).

  14. Automated Forensic Animal Family Identification by Nested PCR and Melt Curve Analysis on an Off-the-Shelf Thermocycler Augmented with a Centrifugal Microfluidic Disk Segment.

    Directory of Open Access Journals (Sweden)

    Mark Keller

    Full Text Available Nested PCR remains a labor-intensive and error-prone biomolecular analysis. Laboratory workflow automation by precise control of minute liquid volumes in centrifugal microfluidic Lab-on-a-Chip systems holds great potential for such applications. However, the majority of these systems require costly custom-made processing devices. Our idea is to augment a standard laboratory device, here a centrifugal real-time PCR thermocycler, with inbuilt liquid handling capabilities for automation. We have developed a microfluidic disk segment enabling an automated nested real-time PCR assay for identification of common European animal groups adapted to forensic standards. For the first time we utilize a novel combination of fluidic elements, including pre-storage of reagents, to automate the assay at constant rotational frequency of an off-the-shelf thermocycler. It provides a universal duplex pre-amplification of short fragments of the mitochondrial 12S rRNA and cytochrome b genes, animal-group-specific main-amplifications, and melting curve analysis for differentiation. The system was characterized with respect to assay sensitivity, specificity, risk of cross-contamination, and detection of minor components in mixtures. 92.2% of the performed tests were recognized as fluidically failure-free sample handling and used for evaluation. Altogether, augmentation of the standard real-time thermocycler with a self-contained centrifugal microfluidic disk segment resulted in an accelerated and automated analysis reducing hands-on time, and circumventing the risk of contamination associated with regular nested PCR protocols.

  15. Diagnóstico de infección congénita por citomegalovirus mediante PCR en gotas de sangre seca en el papel de filtro de la tamización neonatal

    OpenAIRE

    2015-01-01

    Objetivo: Determinar la frecuencia de la infección congénita por citomegalovirus en el grupo de recién nacidos prematuros y/o con restricción de crecimiento intrauterino y establecer asociación entre el diagnóstico de la infección congénita por CMV por PCR en sangre seca y la caracterización de los datos del libro de partos y la base de datos del RUAF. Materiales y Métodos: Es un estudio retrospectivo anónimo no ligado, desarrollado en el hospital la Victoria sede Instituto ...

  16. Detección de toxoplasmosis congénita en líquido amniótico humano mediante la técnica de nested-pcr

    Directory of Open Access Journals (Sweden)

    Ossa H.

    2000-12-01

    Full Text Available

    La toxoplasmosis es provocada por el parasite intracelular obligado Toxoplasma gondii,de la familia Toxoplasmidae (Flores, 1991. Este parasite puede ser asintomático en adultos con un sistema inmune normal, pero puede ser de gran trascendencia en el feto en gestación y en pacientes con SIDA o deficiencia en el sistema inmune (Montoya, 1996. La presencia de anticuerpos antitoxoplasma indica únicamente que la persona se infecto con el microorganismo en un momento dado y no que haya oeste desarrollando la toxoplasmosis necesariamente, pero un resultado positivo indica que el individuo está en riesgo de desarrollar la enfermedad (Perea, 1983. Si la infección se produce durante el embarazo, existe la posibilidad que la toxoplasmosis sea transmitida al feto ocasionando aborto espontaneo, prematuridad o enfermedades severas en el feto, tales como: hidrocefalia y calcificaciones inn-ace- i rebrales (Picazo, 1994. En la mayoría de los casos el diagnóstico biológico de la toxoplasmosis congénita se basa en métodos serológicos indirectos; sin embargo, en los últimos años los diversos estudios realizados en Biología Molecular permitieron utilizar la Técnica de Reacción en Cadena de la Polimerasa (PCR para el diagnóstico de la enfermedad (Hohlfeld, 1994. Los primeros estudios en PCR fueron dirigidos a la amplificación de la secuencia repetitiva del gen B1 de Toxoplasma gondii en líquido amniótico de mujeres infectadas (Grover, 1990. La prueba de PCR en liquido amniótico es definitivamente mas sensible que otras técnicas convencionales usadas, ya que estas presentan dificultad en establecer un diagnóstico segura y oportuno, por esto se ha implementando la técnica de PCR en la detección de la toxoplasmosis, aportando un progreso indiscutible en aquellos casos donde los exámenes clínicos y serológicos presentan limitaciones. También disminuye el tiempo de análisis de las muestras arrojando resultados en un período máximo de 24

  17. Detección mediante RT-PCR en tiempo real del virus vacunal en cerdos inmunizados con la vacuna cubana contra la peste porcina clásica

    Directory of Open Access Journals (Sweden)

    Tania Campos-Cuello

    2016-08-01

    Full Text Available La peste porcina clásica (PPC es una enfermedad viral infectocontagiosa, producida por un virus ARN del género Pestivirus, familia Flaviviridae. En la actualidad es una de las causas de pérdidas económicas en la industria porcina a nivel mundial. En su prevención se han utilizado vacunas vivas atenuadas, empleando la cepa China lapinizada. La Reacción en Cadena de la Polimerasa Reverso Transcriptasa (RT-PCR ha sido uno de los métodos más sensibles aplicado en Medicina Veterinaria para la detección de virus ARN. En el caso del virus de la PPC es muy útil porque el ácido nucleico se puede detectar desde muy temprano en la infección y en periodos más largos en aquellos animales que se recuperan. El objetivo de este estudio fue aplicar la técnica de RT-PCR en tiempo real para la detección de la cepa China lapinizada de la vacuna cubana contra la PPC. Las tonsilas de los cerdos vacunados fueron el órgano más positivo en la detección del ARN del virus vacunal. Los resultados obtenidos evidenciaron una interferencia del virus vacunal en el diagnóstico, siendo el día 12 posvacunación en el que se obtiene una emisión umbral de fluorescencia más bajo.

  18. Analysis of the bacterial diversity existing on animal hide and wool: development of a preliminary PCR-restriction fragment length polymorphism fingerprint database for identifying isolates.

    Science.gov (United States)

    Chen, Yu; Gao, Hongwei; Zhang, Yanming; Deng, Mingjun; Wu, Zhenxing; Zhu, Laihua; Duan, Qing; Xu, Biao; Liang, Chengzhu; Yue, Zhiqin; Xiao, Xizhi

    2012-01-01

    Twenty-one bacterial strains were isolated from imported cattle hide and rabbit wool using two types of media, nutrient broth, and nutrient broth with serum. The bacteria identified were Brevibacillus laterosporus, Leclercia adecarboxylata, Peptococcus niger, Bacillus circulans, Raoultella ornithinolytica, Bacillus subtilis, Bacillus cereus, Bacillus thermobacillus, Bacillus choshinensis, Bacillus sphaericus, Acinetobacter haemolyticus, Sphingomonas paucimobilis, Bacillus thuringiensis, Staphylococcus intermedius, Mycobacteria, Moraxella, Klebsiella pneumoniae, Ralstonia pickettii, Staphylococcus chromogenes, Comamonas testosteroni, and Cupriavidus pauculus. The 16s rDNA gene of each bacterium was amplified using the universal primers 27f and 1492r. The amplicons were digested with AvaI, BamHI, BgII, DraI, EcoRI, EcoRV, HindIII, HinfI, HpaI, PstI, SmaI, TaqII, XbaI, XmaI, AluI, XhoI, and PvuI individually. A specific fingerprint from the PCR-restriction fragment length polymorphism method based on 16s rDNA was obtained for each bacterium. The results showed that the method developed was useful not only for bacterial identification but also for the etiological investigation of pathogens in imported animal hair and wool.

  19. Sensitive Quantification of Aflatoxin B1 in Animal Feeds, Corn Feed Grain, and Yellow Corn Meal Using Immunomagnetic Bead-Based Recovery and Real-Time Immunoquantitative-PCR

    Directory of Open Access Journals (Sweden)

    Dinesh Babu

    2014-12-01

    Full Text Available Aflatoxins are considered unavoidable natural mycotoxins encountered in foods, animal feeds, and feed grains. In this study, we demonstrate the application of our recently developed real-time immunoquantitative PCR (RT iq-PCR assay for sensitive detection and quantification of aflatoxins in poultry feed, two types of dairy feed (1 and 2, horse feed, whole kernel corn feed grains, and retail yellow ground corn meal. Upon testing methanol/water (60:40 extractions of the above samples using competitive direct enzyme linked immunosorbent assay, the aflatoxin content was found to be <20 μg/kg. The RT iq-PCR assay exhibited high antigen hook effect in samples containing aflatoxin levels higher than the quantification limits (0.1–10 μg/kg, addressed by comparing the quantification results of undiluted and diluted extracts. In testing the reliability of the immuno-PCR assay, samples were spiked with 200 μg/kg of aflatoxin B1, but the recovery of spiked aflatoxin was found to be poor. Considering the significance of determining trace levels of aflatoxins and their serious implications for animal and human health, the RT iq-PCR method described in this study can be useful for quantifying low natural aflatoxin levels in complex matrices of food or animal feed samples without the requirement of extra sample cleanup.

  20. Detección y caracterización del virus de bronquitis infecciosa aviaria en Chile mediante RT-PCR y análisis secuencial Detection and characterization of infectious bronchitis virus in Chile by RT-PCR and sequence analysis

    Directory of Open Access Journals (Sweden)

    J C Lopez

    2006-01-01

    Full Text Available Una técnica de reacción en cadena de la polimerasa transcriptasa reversa (RT-PCR junto a una secuenciación fue usada para detectar y caracterizar genéticamente virus diferentes de bronquitis infecciosa aviar (VBIA aislados en Chile. El procedimiento de RT-PCR incluyó el uso de los partidores NT1 y NT2, los cuales se localizaron cerca del término N del gen S1 y cubrieron la región hipervariable. La secuencia amplificada fue alineada y analizada con el programa computacional DNAman, y comparada con secuencias reportadas en GenBank. El nivel de detección de la técnica de RT-PCR fue equivalente al aislamiento viral en huevos cuando se usaron directamente tejidos, pero el ensayo fue más sensitivo cuando fue usado para detectar virus almacenados en fluido alantoideo. Los amplificados de todos los aislados históricos de Chile fueron idénticos en tamaño (193pb y exhibieron entre ellos, al analizar la secuencia una similitud del 71 al 96%. Estos aislados mostraron entre 68 y 97% de similitud con cepas de Estados Unidos, Europa, Asia, Nueva Zelandia y Australia.A reverse transcriptase-polymerase chain reaction (RT-PCR assay, coupled with sequencing, was used to detect and genetically characterize different infectious bronchitis virus (IBV isolates in Chile. The RT-PCR procedure included the use of the primers NT1 and NT2 that were located close to the N-terminus of the S1 gene and bracketed the hypervariable region, and the amplified sequences were aligned and analyzed with DNAman software, and compared with sequences from GenBank. The level of detection of the RTPCR assay was equivalent to virus isolation in eggs when testing tissues directly, but the assay was more sensitive when used to detect virus stored in allantoic fluid. The amplimers from all historical Chilean isolates were identical in size (193 bp and exhibited 71-96% similarity on sequence analysis. These isolates showed between 68-97% similarity to strains from North America

  1. A review of RT-PCR technologies used in veterinary virology and disease control: sensitive and specific diagnosis of five livestock diseases notifiable to the World Organisation for Animal Health

    OpenAIRE

    Hoffmann, Bernd; Beer, Martin; Reid, Scott M.; Mertens, Peter; Oura, Chris A.L.; van Rijn, Piet A.; Slomka, Marek J.; Banks, Jill; Brown, Ian H; Alexander, Dennis J.; King, Donald P.

    2009-01-01

    Abstract Real-time, reverse transcription polymerase chain reaction (rRT-PCR) has become one of the most widely used methods in the field of molecular diagnostics and research. The potential of this format to provide sensitive, specific and swift detection and quantification of viral RNAs has made it an indispensable tool for state-of-the-art diagnostics of important human and animal viral pathogens. Integration of these assays into automated liquid handling platforms for nucleic a...

  2. 检测动物棘球蚴的3种PCR方法的比较%Comparison of 3 PCR methods for detection of Echinococcus spp.in animals

    Institute of Scientific and Technical Information of China (English)

    周正斌; 朱淮民

    2011-01-01

    目的 比较多重PCR(muhiplex PCR)、PCR-限制性片段长度多态性(PCR-RFLP)、半巢式PCR等3种检测棘球蚴的PCR方法,优化可区分多房棘球绦虫、细粒棘球绦虫羊株(G1型)和骆驼株(G6型)、石渠棘球绦虫和带科绦虫的方法.方法 根据细粒棘球绦虫、多房棘球绦虫、石渠棘球绦虫、带科绦虫线粒体基因序列,用DNAStar软件设计半巢式PCR通用引物,建立半巢式PCR法,并与文献报道的多重PCR法、PCR-RFLP平行扩增3份巨颈绦虫囊尾蚴、48份多房棘球蚴、8份细粒棘球蚴样本DNA,比较3种检测方法的特异性和灵敏度,并以细粒棘球蚴组织DNA为模板测定3种方法的灵敏度.结果 半巢式PCR法灵敏度最高,59份样本全部扩增出561 bp条带,但需要测序才能确定虫种;多重PCR法能扩增出3份巨颈绦虫囊尾蚴(带绦虫)样本(3/3,100%),5份(5/8,62.5%)细粒棘球蚴样本,23份(23/48,47.9%)多房棘球蚴样本,其余28份样本为假阴性;PCR-RFLP扩增细粒棘球蚴、多房棘球蚴和巨颈绦虫囊尾蚴(带科绦虫)的阳性率与多重PCR相同,且PCR-RFLP不能区分细粒棘球绦虫G1型和带科绦虫.统计学分析显示半巢式PCR法的棘球蚴检出率高于PCR-RFLP法和多重PCR法.半巢式PCR法灵敏度分别是PCR-RFLP法和多重PCR法的1 000倍和100倍,最低检出细粒棘球蚴DNA浓度为253 pg/lμ.结论 根据不同研究目的,可组合选用多重PCR法、PCR-RFLP法、半巢式PCR法区分细粒棘球绦虫G1型、G6型,多房棘球绦虫,石渠棘球绦虫和带绦虫.%Objective To compare 3 methods(hemi-nested PCR,multiplex PCR and PCR-RFLP),and optimize the method for discrimination of Echinococcus spp.in animals.Methods Hemi-nested PCR primers were designed according to the mitochondria DNA sequence of the Genus Echinococcus with DNAStar software.Fifty-nine samples included E.multilocularis,E.granulosus and Taenia taeniaeformis were collected from Shiqu County.All the samples were

  3. Evaluation of RT-PCR Assay for Routine Laboratory Diagnosis of Rabies in Post Mortem Brain Samples from Different Species of Animals

    OpenAIRE

    Aravindh Babu, R. P.; Manoharan, S.; Ramadass, P.; Chandran, N. D. J.

    2012-01-01

    Rabies in domestic and wild animals continues to be a major public health threat in India. Rapid and accurate diagnosis of rabies in animals is therefore of utmost importance as the individuals who were in contact with the rabid animals are at a greater risk. A significant amount of diagnostic tissue samples submitted to our laboratory are often autolysed and the WHO recommended direct fluorescent antibody test (FAT) for rabies diagnosis cannot be used in such samples. In this pilot study we ...

  4. Assessment of PCR-DGGE for the identification of diverse Helicobacter species, and application to faecal samples from zoo animals to determine Helicobacter prevalence

    DEFF Research Database (Denmark)

    Abu Al-Soud, W.; Bennedsen, M.; On, Stephen L.W.;

    2003-01-01

    bilis and Helicobacter hepaticus in a Nile crocodile, Helicobacter cinaedi in a baboon and a red panda, and Helicobacter felis in a wolf and a Taiwan beauty snake. All of these PCR products (similar to400 bp) showed 100 % sequence similarity to 16S rDNA sequences of the mentioned species. These results...

  5. Molecular Prevalence of Babesia bigemina and Trypanosoma evansi in Dairy Animals from Punjab, India, by Duplex PCR: A Step Forward to the Detection and Management of Concurrent Latent Infections

    Directory of Open Access Journals (Sweden)

    Amrita Sharma

    2013-01-01

    Full Text Available Specific duplex polymerase chain reaction (PCR was employed on 411 (386 cattle and 25 buffaloes blood samples of dairy animals from 9 districts of Punjab, India, for simultaneous detection of Babesia bigemina and Trypanosoma evansi. The results were compared and correlated with conventional Giemsa stained thin blood smear (GSTBS examination and haematological alterations to know the clinical status and pathogenicity of infections. The Bg3/Bg4 and TR3/TR4 primers were used in duplex PCR for B. bigemina and T. evansi amplified products of 689 bp and 257 bp, respectively. The overall prevalence by duplex PCR was found to be 36.49, 2.43, and 3.41% for T. evansi, B. bigemina, and dual infection, respectively. A more significant difference was observed for dual infection status (P≤0.005 as compared to T. evansi (P≤0.05 and B. bigemina (P≤0.01 among various districts under study. A very low prevalence of T. evansi (0.73% and B. bigemina (0.48% was seen by GSTBS. The highly sensitive, specific, and cost-effective duplex PCR was able to detect latent T. evansi and B. bigemina infection in cattle and buffaloes. Haematological evaluation revealed marked pathology in B. bigemina infected group and in dual infected group in contrast to that infected with T. evansi alone.

  6. RT-PCR em pools de soros sangüíneos para o diagnóstico da infecção aguda e de animais persistentemente infectados pelo vírus da diarréia viral bovina RT-PCR in pools of bovine blood serum to detect acute infection and persistently infected animals with bovine viral diarrhea virus

    Directory of Open Access Journals (Sweden)

    D. Pilz

    2007-02-01

    Full Text Available Utilizou-se a técnica da RT-PCR para a detecção da região 5' UTR do genoma do vírus da diarréia viral bovina (BVDV em pools de soros sangüíneos provenientes de um rebanho, constituído por 226 animais, que apresentava distúrbios da reprodução. A partir das amostras individuais de soro e de acordo com a categoria dos animais e o número de animais por categoria foram formados 10 pools (A a J de soros. A primeira avaliação revelou a amplificação de um produto com 290pb nas reações referentes aos grupos D (35 vacas e H (25 bezerros lactentes que, após o desmembramento em amostras individuais, resultou na identificação de 11 vacas lactantes e 12 bezerros em amamentação positivos. Para a identificação de animais persistentemente infectados (PI entre os 23 positivos na primeira avaliação, realizou-se a segunda colheita de soros sangüíneos, três meses após. A RT-PCR das amostras individuais de soro revelou resultado positivo em cinco bezerros. Em dois, foi possível isolar o BVDV em cultivo de células MDBK. A especificidade das reações da RT-PCR foi confirmada pelo seqüenciamento dos produtos amplificados a partir do soro de uma vaca com infecção aguda, de um bezerro PI e das duas amostras do BVDV isoladas em cultivo celular. A utilização da RT-PCR em pools de soros sangüíneos demonstrou ser uma estratégia rápida de diagnóstico etiológico e de baixo custo tanto para a detecção de infecção aguda quanto de animais PI.The 5' untranslated region of the bovine viral diarrhea virus (BVDV genome was detected by RT-PCR assay in pools of blood sera samples collected from a cattle herd (n=226 animals with reproductive failures. Based on the classes of animal and the number of animals per class, the individual blood serum samples were distributed in 10 sera pools (A to J. During the first evaluation a 290bp amplicon was amplified in reactions from groups D (35 cows and H (25 sucking calves. The individual analysis

  7. Simultaneous Detection of Vibrio parahaemolyticus and V .cholerae from Aquatic Animals by Dulplex PCR%水生动物副溶血弧菌、霍乱弧菌的PCR同步检测方法

    Institute of Scientific and Technical Information of China (English)

    2013-01-01

    V ibrio parahaemolyticus and V . cholerae were known to be responsible not only for aquatic animals ,but also to be recognized as pathogenic bacteria for human .In this study ,two pairs of specific primers were designed based on toxR gene for V . parahaemolyticus and lolB gene for V .cholerae ,and simultaneous detection of the both V ibrio was established by a dulplex polymerase chain reaction (PCR) , amplification of 368 bp and 519 bp gene fragments .The results showed that two purposed gene fragments from mixed chromosomal DNA of V . parahaemolyticus and V . cholerae in one PCR reaction were simultaneously amplified by two PCR primers ,and no positive reaction was detected in 5 other controlled pathogenic bacteria , showing strong specificity , and no interference between the two primers . The sensitivity test of the dulplex PCR revealed that the two primers detected V .parahaemolyticus at a level of as few as 1 .75 × 102 cfu/mL ,and V .cholerae at a level of as few as 1 .25 × 102 cfu/mL .The positive amplified results were observed in the 8 aquatic products infected artificially ,and negative amplifi cation in the no-infected 8 aquatic products . The findings prove that the dulplex PCR is a specific and sensitive method for simultaneous detection of the both V ibrio in one PCR reaction ,and can be used as the detection of food safety ,and diagnose of aquatic animals diseases caused by the both V ibrio .%  副溶血弧菌和霍乱弧菌((non-O1/non-O139型),不仅对水产动物具有广泛和强的致病作用,而且危害人类的健康,是人和水产动物共患病的病原菌。以副溶血弧菌的 toxR 基因及霍乱弧菌的lolB 基因设计2对引物,建立了副溶血弧菌和霍乱弧菌的 PCR 同步检测方法,相应的扩增目的片段分别为368 bp 和519 bp 。在 PCR 反应体系中副溶血弧菌和霍乱弧菌可扩增出目的基因片段,5株对照菌无任何扩增条带,表明所建立的双重 PCR 检测方

  8. Application of PCR in Animal Derived Food Safety%PCR在动物源性食品安全方面的应用

    Institute of Scientific and Technical Information of China (English)

    王小建

    2008-01-01

    聚合酶链式反应(polymerase chain reaction,PCR)技术是一项体外酶促扩增DNA 技术,具有特异性强、敏感性高、操作简便、快速高效等特点.在肉品科学方面有潜在的应用价值,本文综述PCR在肉类科学中的应用:如微生物的检测,肉类品种的鉴定等.

  9. Identification of new flagellin-encoding fliC genes in Escherichia coli isolated from domestic animals using RFLP-PCR and sequencing methods Identificação de novas flagelinas codificadas por fliC em Escherichia coli isoladas de animais domésticos utilizando RFLP-PCR e sequenciamento

    Directory of Open Access Journals (Sweden)

    Cláudia de Moura

    2013-04-01

    Full Text Available Identification of Escherichia coli requires knowledge regarding the prevalent serotypes and virulence factors profiles allows the classification in pathogenic/non-pathogenic. However, some of these bacteria do not express flagellar antigen invitro. In this case the PCR-restriction fragment length polymorphism (RFLP-PCR and sequencing of the fliC may be suitable for the identification of antigens by replacing the traditional serology. We studied 17 samples of E. coli isolated from animals and presenting antigen H nontypeable (HNT. The H antigens were characterized by PCR-RFLP and sequencing of fliC gene. Three new flagellin genes were identified, for which specific antisera were obtained. The PCR-RFLP was shown to be faster than the serotyping H antigen in E. coli, provided information on some characteristics of these antigens and indicated the presence of new genes fliC.A identificação da Escherichia coli requer conhecimento sobre os sorotipos e fatores de virulência prevalentes permitindo a classificação em patogênico/não patogênico. No entanto, algumas destas bactérias não expressam o antígeno flagelar in vitro. Neste caso, o PCR-restriction fragment length polymorphism (RFLP-PCR e o sequenciamento do gene fliC podem ser adequados para a identificação desses antígenos, substituindo a sorologia tradicional. Nesta pesquisa foram estudadas 17 amostras de E. coli isoladas de animais e que apresentavam antígeno H não tipável (HNT. Os antígenos H foram caracterizados por PCR-RFLP e sequenciamento do gene fliC. Três novos genes da flagelina foram identificados, para os quais anti-soros específicos foram obtidos. A técnica PCR-RFLP mostrou-se mais rápida que a sorotipagem do antígeno H em E. coli, fornecendo informações sobre algumas características desses antígenos e indicou a presença de novos genes fliC.

  10. A review of RT-PCR technologies used in veterinary virology and disease control: sensitive and specific diagnosis of five livestock diseases notifiable to the World Organisation for Animal Health.

    Science.gov (United States)

    Hoffmann, Bernd; Beer, Martin; Reid, Scott M; Mertens, Peter; Oura, Chris A L; van Rijn, Piet A; Slomka, Marek J; Banks, Jill; Brown, Ian H; Alexander, Dennis J; King, Donald P

    2009-10-20

    Real-time, reverse transcription polymerase chain reaction (rRT-PCR) has become one of the most widely used methods in the field of molecular diagnostics and research. The potential of this format to provide sensitive, specific and swift detection and quantification of viral RNAs has made it an indispensable tool for state-of-the-art diagnostics of important human and animal viral pathogens. Integration of these assays into automated liquid handling platforms for nucleic acid extraction increases the rate and standardisation of sample throughput and decreases the potential for cross-contamination. The reliability of these assays can be further enhanced by using internal controls to validate test results. Based on these advantageous characteristics, numerous robust rRT-PCRs systems have been developed and validated for important epizootic diseases of livestock. Here, we review the rRT-PCR assays that have been developed for the detection of five RNA viruses that cause diseases that are notifiable to the World Organisation for Animal Health (OIE), namely: foot-and-mouth disease, classical swine fever, bluetongue disease, avian influenza and Newcastle disease. The performance of these tests for viral diagnostics and disease control and prospects for improved strategies in the future are discussed.

  11. Improved Culture Medium (TiKa) for Mycobacterium avium Subspecies Paratuberculosis (MAP) Matches qPCR Sensitivity and Reveals Significant Proportions of Non-viable MAP in Lymphoid Tissue of Vaccinated MAP Challenged Animals.

    Science.gov (United States)

    Bull, Tim J; Munshi, Tulika; Mikkelsen, Heidi; Hartmann, Sofie B; Sørensen, Maria R; Garcia, Joanna S; Lopez-Perez, Paula M; Hofmann, Sven; Hilpert, Kai; Jungersen, Gregers

    2016-01-01

    The quantitative detection of viable pathogen load is an important tool in determining the degree of infection in animals and contamination of foodstuffs. Current conventional culture methods are limited in their ability to determine these levels in Mycobacterium avium subspecies paratuberculosis (MAP) due to slow growth, clumping and low recoverability issues. The principle goal of this study was to evaluate a novel culturing process (TiKa) with unique ability to stimulate MAP growth from low sample loads and dilutions. We demonstrate it was able to stimulate a mean 29-fold increase in recoverability and an improved sensitivity of up to three logs when compared with conventional culture. Using TiKa culture, MAP clumping was minimal and produced visible colonies in half the time required by standard culture methods. Parallel quantitative evaluation of the TiKa culture approach and qPCR on MAP loads in tissue and gut mucosal samples from a MAP vaccine-challenge study, showed good correlations between colony counts (cfu) and qPCR derived genome equivalents (Geq) over a large range of loads with a 30% greater sensitivity for TiKa culture approach at low loads (two logs). Furthermore, the relative fold changes in Geq and cfu from the TiKa culture approach suggests that non-mucosal tissue loads from MAP infected animals contained a reduced proportion of non-viable MAP (mean 19-fold) which was reduced significantly further (mean 190-fold) in vaccinated "reactor" calves. This study shows TiKa culture equates well with qPCR and provides important evidence that accuracy in estimating viable MAP load using DNA tests alone may vary significantly between samples of mucosal and lymphatic origin.

  12. Non-invasive assessment of animal exercise stress: real-time PCR of GLUT4, COX2, SOD1 and HSP70 in avalanche military dog saliva.

    Science.gov (United States)

    Diverio, S; Guelfi, G; Barbato, O; Di Mari, W; Egidi, M G; Santoro, M M

    2015-01-01

    Exercise has been shown to increase mRNA expression of a growing number of genes. The aim of this study was to assess if mRNA expression of the metabolism- and oxidative stress-related genes GLUT4 (glucose transporter 4), COX2 (cyclooxygenase 2), SOD1 (superoxide dismutase 1) and HSP70 (heat shock protein 70) in saliva changes following acute exercise stress in dogs. For this purpose, 12 avalanche dogs of the Italian Military Force Guardia di Finanza were monitored during simulation of a search for a buried person in an artificial avalanche area. Rectal temperature (RT) and saliva samples were collected the day before the trial (T0), immediately after the descent from a helicopter at the onset of a simulated avalanche search and rescue operation (T1), after the discovery of the buried person (T2) and 2 h later (T3). Expressions of GLUT4, SOD1, COX2 and HSP70 were measured by real-time PCR. The simulated avalanche search and rescue operation was shown to exert a significant effect on RT, as well as on the expression of all metabolism- and oxidative stress-related genes investigated, which peaked at T2. The observed expression patterns indicate an acute exercise stress-induced upregulation, as confirmed by the reductions in expression at T3. Moreover, our findings indicate that saliva is useful for assessing metabolism- and oxidative stress-related genes without the need for restraint, which could affect working dog performance.

  13. Investigation of Leptospira infection in three new experimental animals by PCR methods%应用 PCR 方法对三种新型实验动物钩端螺旋体感染情况的调查研究

    Institute of Scientific and Technical Information of China (English)

    冯育芳; 邢进; 巩薇; 岳秉飞; 贺争鸣

    2014-01-01

    目的:建立有效的钩端螺旋体 PCR 检测方法,并对树鼩((tree shrew, Tupaia belangeri))、长爪沙鼠( Meriones unguiculatus; Mongolian gerbil)和灰仓鼠( Cricetulus migratorius)等三种新型实验动物进行感染情况调查。方法针对 NCBI 公布的钩端螺旋体序列,设计并筛选特异性引物,优化 PCR 体系,进行特异性和敏感性测试;并运用优化 PCR 方法对树鼩、长爪沙鼠和灰仓鼠样品进行检测。结果成功建立钩端螺旋体 PCR 检测方法,序列测定验证了该方法的特异性。普通级树鼩钩端螺旋体的阳性率为8.33%,普通级长爪沙鼠钩端螺旋体为100%,清洁级长爪沙鼠和清洁级灰仓鼠钩端螺旋体的阳性率为0%。结论本研究建立了钩端螺旋体 PCR 检测方法,调查了树鼩、长爪沙鼠和灰仓鼠三种新型实验动物的感染情况,为这三种实验动物的研究和使用奠定基础。%Objective To establish an effective PCR assay for leptospirosis detection , and applicate the assay in tree shrew, mongolian gerbil and gray hamster .Methods Sequence of leptospira was obtained from the NCBI Genbank , and primers were designed based on the sequences .The positive amplified fragments were sequenced to verify the reliability of the method.The samples from tree shrew, mongolian gerbils and hamsters were tested using this PCR method .Results The PCR method for detection of leptospirosis was successfully established .The positive rate of Leptospira was 8.33% in 60 samples of conventional tree shrews , 100% in 104 samples of the conventional Mongolian gerbils , and 0% in 60 samples of clean gray hamsters.Conclusions The establishment of this PCR assay is useful in the detection of leptospirosis in tree shrew, mongolian gerbil and gray hamster .The results of our investigation of leptospira infection levels of the three new experimental animals may promote their application in biomedical research .

  14. PCR detection of pathogenic Vibrio and Aeromonas hydrophila in aquaculture animals in flowing-through systems%流水式养殖动物中弧菌和嗜水气单胞菌的PCR法监测

    Institute of Scientific and Technical Information of China (English)

    傅松哲; 夏文; 涂俊凌; 樊国印; 刘鹰

    2011-01-01

    To investigate the prevalence of pathogenic Vibrio and Aeromonas hydrophila in aquaculture animals in flowing through systems in Nanchang, animals were sampled over an 18-month period between March 2009 and September 2010. Seven pairs o{ primers were designed for toxin sub-unit A(ctxA) gene, O139 special gene, O1 special gene, tcpA gene, toxR gene, haemolytic gene, vvhA gene and aerolysin(aerA) gene. Specific PCR was established to simultaneously detect these 7 genes. Strains of Vibrio cholerae, V. parahaernolyticus, V.vulnificus and Aerornonas hydrophila can be detected and differentiated by the size and numbers of specific PCR products through electrophoresis bands examination. In 1 440 tested samples, 17 samples were Vibrio parahaernolyticus positive (1.74%), 25 samples were Vibrio cholerae positive (1.18% ), and no Aeromonas hydrophila and Vibrio vulnificus were found in the samples. The results of PCR detection were consistent with those of the traditional culture methods. The results indicated that the levels of pathogenic Vibrio in Nanchang area were very low, while the levels of pathogenic Vibrio in Amyda Sincnsis and Rana catesbeiana were relatively high.%为了解流水式养殖场养殖动物中弧菌和气单胞菌的感染状况,分别针对霍乱孤菌肠毒素A亚单位(ctxA)基因、0139和01群群特异基因、毒力协同调节菌毛A亚单位基因(tcpA)、中心调节蛋白基因(toxR)、副溶血弧菌不耐热溶血素基因(tlh)、创伤弧菌Vibrio vulnificus溶细胞素基因(vvhA)和嗜水气单胞菌的气溶素基因(aerA)设计引物,建立了聚合酶链式反应(PCR)检测技术,PCR产物经电泳,根据扩增条带的大小和数目,检测和区分霍乱孤菌、嗜水气单胞菌和副溶血孤菌.对南昌沿岸霍乱流行水域及附近养殖场的养殖动物进行了18个月的连续监测,对1 440份水产品的增菌液进行PCR检测,17份检出副溶血弧菌,25份检出霍乱弧菌,创伤弧菌和嗜水气单胞菌

  15. 实验动物皮肤病原真菌多重 PCR 检测方法的建立与初步应用%Development and application of a multiplex PCR assay for pathogenic dermatophytes in laboratory animals

    Institute of Scientific and Technical Information of China (English)

    邢进; 冯育芳; 岳秉飞; 贺争鸣

    2015-01-01

    目的:建立石膏样毛癣菌(Tm)、石膏样小孢子菌(Mg)、犬小孢子菌(Mc)和猴类毛癣菌(Am)的多重PCR检测方法,用于实验动物皮肤病原真菌的快速检测。方法根据Genbank公布的四种皮肤病原真菌的18S-28S rRNA序列设计特异性引物,通过引物、dNTP、TaqDNA聚合酶浓度及退火温度和时间的优化,建立四种皮肤病原真菌的多重PCR体系。经特异性和敏感性检验后,对15份人工感染真菌大鼠毛和260份实验动物毛发样本进行检测。结果所建多重PCR方法能够有效区分四种皮肤病原真菌,产生192 bp( Tm)、460 bp( Mg)、290 bp( Mc)和602 bp( As)的目的片段,最低检测限分别为5.9 pg/μL、6.6 pg/μL、9.5 pg/μL和5.1 pg/μL。被检的15份人工感染样本扩增结果准确,260份毛发样本中未检测出四种真菌。结论所建立的多重PCR方法能够同时对实验动物中的四种皮肤病原真菌进行快速的检测,具有较高的应用价值。%Objective To develop a multiplex polymerase chain reaction ( mPCR) assay for detection of four pathogenic dermatophytes [Trichophyton mentagrophytes (Tm), Microsporum gypseum (Mg), Microsporum canis (Mc), and Arthroderma simii ( As) ] in laboratory animals, which could be used rapidly and simultaneously for direct detection of those four pathogens.Methods We designed 5 specific primers according to 18S-28S rRNA sequences of the four pathogenic dermatophytes reported in Genbank. The four mPCR assays were established through optimizing the concentration of primers, dNTP, TaqDNA polymerase and the annealing temperature.After verifying the specificity and sensibility, this method was used to detect 15 hair samples with artificial infection and 260 samples taken from laboratory animals.Results This mPCR technique can distinguish the four dermatophytes by producing 192 bp( Tm) ,460 bp( Mg) , 290 bp( Mc) and 602 bp( As) fragments

  16. PCR thermocycler

    Science.gov (United States)

    Benett, William J.; Richards, James B.

    2003-01-01

    A sleeve-type silicon polymerase chain reaction (PCR) chamber or thermocycler having improved thermal performance. The silicon sleeve reaction chamber is improved in thermal performance by etched features therein that reduce thermal mass and increase the surface area of the sleeve for cooling. This improved thermal performance of the thermocycler enables an increase in speed and efficiency of the reaction chamber. The improvement is accomplished by providing grooves in the faces of the sleeve and a series of grooves on the interior surfaces that connect with grooves on the faces of the sleeve. The grooves can be anisotropically etched in the silicon sleeve simultaneously with formation of the chamber.

  17. 5种外来动物疫病并联荧光定量PCR检测方法的研究%Development of Parallel Fluorescene Quantitative PCR for Detection of the Five Exotic Animal Diseases

    Institute of Scientific and Technical Information of China (English)

    于恒智; 刘晔; 程玮; 张旭东; 苗富春; 赵丹; 张守峰; 刘芳伊; 牛文博

    2016-01-01

    According to the gene sequences of related viruses reported in Genbank,primers and probes were designed based on the conservative region of Peste des petits ruminants virus(PPRV)African swine fever virus(ASFV), West Nile fever virus(WNV),Hendra virus(HeV)and Nipah virus(NiV)(the conservative regions were N gene of PPRV,p72 gene of ASFV,H gene of HEV and NIV,and PRM gene of WNV,respectively). After screening a set of specific primers and a TaqMan probe,a real-time fluorescene quantitative PCR for detection of the above five exotic animal diseases was established. The clinical detection results of 103 samples used this method showed that all the samples were tested negative except the positive control,which verified the parallel fluorescene quantitative PCR of Taqman mode established in this study has characterisitics of differential diagnosis. The method can be used as a reference method for clinical detection of above-mentioned five kinds of pathogens.%根据 GenBank中发表的相关病毒的基因序列,通过分析比较小反刍兽疫病毒(PPRV)、非洲猪瘟病毒(ASFV)、西尼罗河热病毒(WNV)、亨德拉病毒(HeV)和尼帕病毒(NiV)的保守区域(PPRV的N基因、ASFV的P72基因、HeV和NiV的H基因,以及WNV的PrM基因),各设计筛选出一套特异性引物和Taqman探针,建立了5种外来动物疫病并联荧光定量RT-PCR检测方法。利用该方法,对103份样本进行了临床检测,结果除阳性对照外,其他均为阴性。检测结果验证了本研究建立的Taqman模式并联荧光定量PCR检测方法具有鉴别诊断的特点,可以作为临床检测这5种病原体的参考方法。

  18. Tratamiento del hombro doloroso mediante terapia manual

    OpenAIRE

    Gabucio López, Pedro

    2008-01-01

    Introducción: Las disfunciones de hombro son un problema de salud común en las sociedades occidentales. Algunos protocolos de tratamiento han sido desarrollados mediante ensayos clínicos con pacientes que presentan dolor de hombro. Sin embargo, no hay evidencias que sustenten que un protocolo es mejor que otros. El principal objetivo de este trabajo es presentar un caso clínico en el que mediante terapia manual y la prescripción de ejercicios físicos se consigue la resolución del ...

  19. Research Progress and Applications of PCR Technology on Diseases Diagnose in Aquaculture Animal%PCR技术在水产养殖动物疾病诊断中的应用研究进展

    Institute of Scientific and Technical Information of China (English)

    冯腾; 王秀利; 常亚青

    2006-01-01

    聚合酶链式反应(Polymerase Chain Reaction,PCR)因具有特异性强、敏感性高、快速、简便等特点被应用于水产养殖动物的疾病诊断.目前所使用的大多是PCR及其所衍生出来的技术,主要有常规PCR、反转录PCR(reverse transcription-polymerase chain reaction,RT-PCR)、定量PCR(quantitative polymerase chain reaction)、嵌套式PCR(nested polymerase chain reaction)、多重PCR(multiplex polymerase chain reaction)、连环恒温扩增技术(loopmediated isothermal amplification,LAMP),及其与其他技术的联合应用.就国内外的研究情况,对上述技术的原理、特点、应用等方面进行了综述.

  20. Reverse-transcription PCR (RT-PCR).

    Science.gov (United States)

    Bachman, Julia

    2013-01-01

    RT-PCR is commonly used to test for genetic diseases and to characterize gene expression in various tissue types, cell types, and over developmental time courses. This serves as a form of expression profiling, but typically as a candidate approach. RT-PCR is also commonly used to clone cDNAs for further use with other molecular biology techniques (e.g., see Oligo(dT)-primed RT-PCR isolation of polyadenylated RNA degradation intermediates and Circularized RT-PCR (cRT-PCR): analysis of RNA 5' ends, 3' ends, and poly(A) tails).

  1. Improved Culture Medium (TiKa) for Mycobacterium avium Subspecies Paratuberculosis (MAP) Matches qPCR Sensitivity and Reveals Significant Proportions of Non-viable MAP in Lymphoid Tissue of Vaccinated MAP Challenged Animals

    DEFF Research Database (Denmark)

    Bull, Tim J.; Munshil, Tulika; Melvang, Heidi Mikkelsen

    2017-01-01

    in recoverability and an improved sensitivity of up to three logs when compared with conventional culture. Using TiKa culture, MAP clumping was minimal and produced visible colonies in half the time required by standard culture methods. Parallel quantitative evaluation of the TiKa culture approach and qPCR on MAP...... loads in tissue and gut mucosal samples from a MAP vaccine-challenge study, showed good correlations between colony counts (cfu) and qPCR derived genome equivalents (Geq) over a large range of loads with a 30% greater sensitivity for TiKa culture approach at low loads (two logs). Furthermore......Ka culture equates well with qPCR and provides important evidence that accuracy in estimating viable MAP load using DNA tests alone may vary significantly between samples of mucosal and lymphatic origin....

  2. A review of RT-PCR technologies used in veterinary virology and disease control: sensitive and specific diagnosis of five livestock diseases notifiable to the World Organisation for Animal Health

    NARCIS (Netherlands)

    Hoffmann, B.; Beer, M.; Reid, S.M.; Mertens, P.; Oura, C.A.L.; Rijn, van P.A.; Slomka, M.J.; Banks, J.; Brown, I.H.; Alexander, D.J.; King, D.P.

    2009-01-01

    Real-time, reverse transcription polymerase chain reaction (rRT-PCR) has become one of the most widely used methods in the field of molecular diagnostics and research. The potential of this format to provide sensitive, specific and swift detection and quantification of viral RNAs has made it an indi

  3. PCR技术在水产养殖动物病原微生物检测中的应用%The Application of PCR in Detecting for Aquatic Animal's Pathogentic Microorganism.

    Institute of Scientific and Technical Information of China (English)

    李志明; 冯伟业

    2007-01-01

    随着我国水产养殖集约化程度不断提高,虾类、贝类和鱼类养殖业受到病原微生物的严重影响.因此,如何快速准确预测和诊断水产动物疾病,成为当前水产养殖业十分重要而突出的问题.近年来,随着PCR技术的介入,带动了水产养殖动物病原微生物的检测技术快速发展.文章介绍了PCR及其衍生技术(RT-PCR,Real-time PCR,免疫PcR,套式引物PCR,多重PCR)的原理、特点及在水产养殖动物病原微生物检测中的应用,并对水产养殖动物病原微生物诊断技术的发展方向做出了展望.

  4. Sex Determination Using PCR

    Science.gov (United States)

    Kima, Peter E.; Rasche, Madeline E.

    2004-01-01

    PCR has revolutionized many aspects of biochemistry and molecular biology research. In the following exercise, students learn PCR by isolating their own DNA, amplifying specific segments of the X and Y chromosomes, and estimating the sizes of the PCR products using agarose gel electrophoresis. Based on the pattern of PCR products, students can…

  5. Species identification of processed animal proteins (PAPs) in animal feed containing feed materials from animal origin.

    Science.gov (United States)

    Axmann, Sonja; Adler, Andreas; Brandstettner, Agnes Josephine; Spadinger, Gabriela; Weiss, Roland; Strnad, Irmengard

    2015-01-01

    Since June 2013 the total feed ban of processed animal proteins (PAPs) was partially lifted. Now it is possible to mix fish feed with PAPs from non-ruminants (pig and poultry). To guarantee that fish feed, which contains non-ruminant PAPs, is free of ruminant PAPs, it has to be analysed with a ruminant PCR assay to comply with the total ban of feeding PAPs from ruminants. However, PCR analysis cannot distinguish between ruminant DNA, which originates from proteins such as muscle and bones, and ruminant DNA, which comes from feed materials of animal origin such as milk products or fat. Thus, there is the risk of obtaining positive ruminant PCR signals based on these materials. The paper describes the development of the combination of two analysis methods, micro-dissection and PCR, to eliminate the problem of 'false-positive' PCR signals. With micro-dissection, single particles can be isolated and subsequently analysed with PCR.

  6. Propidium monoazide reverse transcription PCR and RT-qPCR for detecting infectious enterovirus and norovirus

    Science.gov (United States)

    Presently there is no established cell line or small animal model that allows for the detection of infectious human norovirus. Current methods based on RT-PCR and RT-qPCR detect both infectious and non-infectious virus and thus the conclusions that may be drawn regarding the publ...

  7. Application of real-time fluorescence and quantitative PCR in detecting influenza A virus in animals transported by railway%实时荧光定量PCR在铁路运输动物携带甲型流感病毒检测中的应用

    Institute of Scientific and Technical Information of China (English)

    区伟全; 卢国钧

    2011-01-01

    目的 探讨应用实时荧光PCR对铁路运输动物携带甲型流感病毒进行快速检测的可行性,为制定铁路系统有效的流感监测方案奠定基础.方法 采集死禽咽喉部和泄殖腔拭子,活体动物用拭子涂抹泄殖腔外周,粪便和毛用拭子涂抹表面.依据、甲型流感病毒核酸检测试剂盒(PCR-荧光探针法)说明书以及ABI 7500F型实时荧光定量PCR的SDS软件操作.结果 共采集广州中铁快运货场动物样本250份,检出甲型流感病毒核酸阳性11份,阳性率4.4%,其中以鸽类样本的阳性检出率最高,鹅次之,鸭最低.结论 实时荧光PCR检测方法适合铁路运输的实际要求,结合进一步病毒分型和病毒基因测序,对掌握病毒流行趋势,做好预防控制工作具有重要意义.%Objective The feasibility of rapid detection of influenza A virus in animals transported by railway using real-time fluorescent PCR is discussed, and to lay the foundation for working out an effective influenza monitoring plan in railway system. Methods Throat and cloacal swab were collected from dead birds. For living animals, peripheral parts of cloaca smears and coat and manure swabs were collected. Influenza A virus was detected by real-time fluorescence quantitative PCR using ABI 7500 F-type real-time fluorescence quantitative PCR and the result was analyzed by the SDS software and followed the “national influenza monitoring scheme”. Results 250 animal samples were collected in the freight yard of Guangzhou Railway Express, and detected for the nucleic acid of influenza A virus. 11 were positive, with a positive rate of 4.4%.Conclusion Real-time fluorescent PCR detection method meets the practical need of rail transport. It is important to grasp the trend of virus prevalent for prevention and control, combining with virus typing and viral gene sequencing.

  8. Development of a combined canine distemper virus specific RT-PCR protocol for the differentiation of infected and vaccinated animals (DIVA) and genetic characterization of the hemagglutinin gene of seven Chinese strains demonstrated in dogs.

    Science.gov (United States)

    Yi, Li; Cheng, Shipeng; Xu, Hongli; Wang, Jianke; Cheng, Yuening; Yang, Shen; Luo, Bin

    2012-01-01

    A combined reverse-transcription polymerase chain reaction (RT-PCR) method was developed for the detection and differentiation of wild-type and vaccine strains of the canine distemper virus (CDV). A pair of primers (P1/P2) was used to detect both CDV wild-type strains and vaccines. Another pair (P3/P4) was used to detect only CDV wild-type strains. A 335bp fragment was amplified from the genomic RNA of the vaccine and wild-type strains. A 555bp fragment was amplified specifically from the genomic RNA of the wild-type strains. No amplification was achieved for the uninfected cells, cells infected with canine parvovirus, canine coronavirus, or canine adenovirus. The combined RT-PCR method detected effectively and differentiated the CDV wild-type and vaccine strains by two separate RT-PCRs. The method can be used for clinical detection and epidemiological surveillance. The phylogenetic analysis of the hemagglutinin gene of the local wild-type CDV strains revealed that the seven local isolates all belonged to the Asia-1 lineage, and were clustered closely with one another at the same location. These results suggested that the CDV genotype Asia-1 is circulating currently in domestic dogs in China.

  9. Establishment of multiplex fluorescent PCR assay for the detection of bovine,goat and sheep- derived materials in animal products and feeding stuffs%动物产品及饲料中牛源和羊源性成分三重荧光PCR检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    赵冉; 蔡振鸿; 陈永锋

    2012-01-01

    为鉴定和区分饲料及动物产品中牛、山羊、绵羊源性成分,根据线粒体DNA(mitochondrial DNA,mtDNA)种间保守序列,设计合成了3对特异性引物与TaqMan探针,通过对荧光PCR反应体系和反应条件的优化筛选,建立了三重荧光PCR方法,在同一个荧光PCR反应中完成3种动物源性成分的检测.用该方法对16种不同源性的动物DNA进行检测,结果表明能特异地鉴别检测出牛、山羊和绵羊源性成分,且敏感性比现行国标PCR法高100倍.该方法适用于饲料、肉制品、奶制品等动物源性产品的检测.%In this study, a multiplex fluorescent PCR assay capable of detecting and differentiating bovine, goat and sheep-derived materials was developed using specific primers and TaqMan probes designed according to mitochondrial DNA gene sequences. The multiplex fluorescent PCR reaction system was optimized. The developed method could successfully detect bovine, goat and sheep-derived materials of 16 animals. The result showed that the real-time PCR can work accurately and efficiently, and can be 100 times as sensitive as conventional PCR of GB/T 20190-2006. Therefore, this method has the potential application for detection of bovine, goat and sheep-derived materials in feed, meat products, milk products, etc.

  10. Animal research

    DEFF Research Database (Denmark)

    Olsson, I.A.S.; Sandøe, Peter

    2012-01-01

    in research is analyzed from the viewpoint of three distinct ethical approaches: contractarianism, utilitarianism, and animal rights view. On a contractarian view, research on animals is only an ethical issue to the extent that other humans as parties to the social contract care about how research animals......This article presents the ethical issues in animal research using a combined approach of ethical theory and analysis of scientific findings with bearing on the ethical analysis. The article opens with a general discussion of the moral acceptability of animal use in research. The use of animals...... are faring. From the utilitarian perspective, the use of sentient animals in research that may harm them is an ethical issue, but harm done to animals can be balanced by benefit generated for humans and other animals. The animal rights view, when thoroughgoing, is abolitionist as regards the use of animals...

  11. 动物产品中猪源性和牛源性成分双重荧光PCR检测方法的建立%Identification of Swine-derived and Bovine-derived Materials in Animal Products by Multiplex Fluorescent PCR Assay

    Institute of Scientific and Technical Information of China (English)

    赵冉

    2012-01-01

    [Objective ] To develop a multiplex fluorescent PCR assay method to detect swine-derived and bovine-derived materials in animal products. [ Method ] Two pairs of specific primers and two fluorescent-labled probes were designed and synthesized according to mitochondrial DNA gene sequences. The multiplex fluorescent PCR reaction system was established through the optimized screening of reaction system and reaction conditions, and swine-derived and bovine-derived materials in animal products were detected in the same fluorescent PCR reaction system. Finally, the sensitivity and specificity of the assay were evaluated. [ Result ] The method successfully detected swine-derived and bovine-derived materials from 16 animals. Swine-derived and bovine-derived materials could obtain responsible S-type amplification curve, while there was no amplification curve from the other 14 animals. The detection limits of the assay for swine and bovine were either up to 10^-5, which was consistent with that of single-weight fluorescent PCR assay method. [Conclusion ] The multiplex fluorescent PCR developed in the study has strong specificity and good sensitivity, so it is suitable for the detection of swine-derived and bovine-derived materials in feed, meat products, milk products, etc..%[目的]建立双重荧光PCR法,检测动物产品中猪、牛源性成分。[方法]分别针对猪、牛线粒体DNA(mitochondrial DNA,mtDNA)种间保守基因,设计特异性引物与探针,通过对反应体系和反应条件的优化筛选,建立了双重荧光PCR方法,在同一个荧光PCR反应中完成2种动物源性成分的检测。并对该方法的特异性、敏感性进行评估。[结果]对16种不同的动物DNA进行检测.仅猪、牛源性成分收集到相应的典型的S型扩增曲线,对猪、牛二联模板的检测,可同时收集到相应模板的扩增曲线.其余14种动物源性成分未发现扩增曲线。双重荧光PCR对猪肉、牛肉模

  12. Toxoplasmosis in wild and domestic animals

    Science.gov (United States)

    Toxoplasma gondii is widely distributed in wild and domestic animals. The present chapter reviews toxoplasmosis in wild and domestic animals. Coverage in wild animal species is limited to confirmed cases of toxoplasmosis, cases with parasite isolation, cases with parasite detection by PCR, and exper...

  13. Animal Bites

    Science.gov (United States)

    ... or territory. Attacks by pets are more common. Animal bites rarely are life-threatening, but if they become infected, you can develop serious medical problems. To prevent animal bites and complications from bites Never pet, handle, ...

  14. Animal Farm

    Institute of Scientific and Technical Information of China (English)

    徐蓉蓉

    2015-01-01

    This essay first introduce the background of Animal Farm and a brief introduction of the author.Then it discuss three thesis about this novel and briefly discussed about it.At last it give highly review on Animal Farm.

  15. Animal models

    DEFF Research Database (Denmark)

    Gøtze, Jens Peter; Krentz, Andrew

    2014-01-01

    In this issue of Cardiovascular Endocrinology, we are proud to present a broad and dedicated spectrum of reviews on animal models in cardiovascular disease. The reviews cover most aspects of animal models in science from basic differences and similarities between small animals and the human...... pathology, to biomarkers in diagnosis and prognostic evaluation, to drug testing and targeted medicine....

  16. Animal Deliberation

    NARCIS (Netherlands)

    Driessen, C.P.G.

    2014-01-01

    While much has been written on environmental politics on the one hand, and animal ethics and welfare on the other, animal politics, as the interface of the two, is underexamined. There are key political implications in the increase of animal protection laws, the rights of nature, and political parti

  17. Entry, Descent, Landing Animation (Animation)

    Science.gov (United States)

    2005-01-01

    [figure removed for brevity, see original site] Click on the image for Entry, Descent, Landing animation This animation illustrates the path the Stardust return capsule will follow once it enters Earth's atmosphere.

  18. Animal Shelter

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Beijing activist Zhang Luping gives up a lucrative business career to provide a home for stray and abandoned pets "I have never been married, but I have I hundreds of children," said Zhang Luping, founder of the Beijing Human and Animal Environment Education Center (the Animal Center). "God sent me to this planet and gave me the mission of taking care of helpless and homeless dogs and cats. I will never let Him down." The Animal Center, one of a few non-

  19. Animal ethics

    DEFF Research Database (Denmark)

    Palmer, Clare; Sandøe, Peter

    2011-01-01

    This chapter describes and discusses different views concerning our duties towards animals. First, we explain why it is necessary to engage in thinking about animal ethics and why it is not enough to rely on feelings alone. Secondly, we present and discuss five different kinds of views about...... the nature of our duties to animals. They are: contractarianism, utilitarianism, the animal rights view, contextual views, and a respect for nature view. Finally, we briefly consider whether it is possible to combine elements from the presented views, and how to make up one’s mind....

  20. Animated Asphalt

    DEFF Research Database (Denmark)

    Paldam, Camilla Skovbjerg

    2015-01-01

    “animation”, defined as “an innate (and learnable) ability of our bodies to discover life in inanimate images” (Belting 2012, 188). In this essay I investigate the animation of pictures in dialogue with Mitchell, both by addressing general questions such as: how is animation of otherwise static pictures...... to be understood? How does animation differ in different media? And in particular by focusing on and questioning the gender positions inherent in Mitchell’s theory. Animation has an erotic component of seduction and desire, and what pictures want, becomes for Mitchell, what women want. There is of course no simple...

  1. Development and Application of Multiplex PCR Assay for the Detection of Campylobacter jejuni in Animal Origin Food%动物性食品源空肠弯曲杆菌二重PCR检测方法的建立及应用

    Institute of Scientific and Technical Information of China (English)

    陈荀; 张晓利; 刘书亮

    2011-01-01

    According to the 16S rDNA and hip O gene sequences of C.jejuni in GenBank, two pairs of specific primers were designed and used in multiplex PCR for detection of C.jejuni in animal origin food.Multiplex PCR was developed and applied to the sample testing.The results indicated that two specific fragments (699bp and 366bp) were detected after amplification of the DNA template of C.jejuni, while other bacteria strains ( 11 species tested) were not detected.Meanwhile, the 16S rDNA and hip O gene sequence of C.jejuni ATCC33560 exhibited a high similarity with those of some strains of C.jejuni presented in GenBank.The total assay could be completed in 27h with a detection limit of 2.4 ~ 16CFU/mL.The chicken, pork, beef and milk from Ya'an markets in Sichuan province were detected by the multiplex PCR, and 38.0% , 28.3% , 17.1% and 8.6% of them were found respectively to be positive for C.jejuni.Multiplex PCR assay was specific, sensitive and time - saving, which provided reference for detection of C.jejuni in animal origin food.%根据GenBank空肠弯曲杆菌(Campylobacterjejuni,Cj)的16S rDNA及hip O(编码马尿酸酶基因)序列设计两对特异引物,建立检测动物性食品源Cj的二重PCR方法,并应用于样品检测.结果显示只对Cj能特异的扩增出699bp和366bp两个基因片段,而大肠杆菌、沙门氏菌等其他11种细菌均未扩增出条带;Cj标准株ATCC33560的16S rDNA及hip O序列与GenBank其他Cj的相应序列具高度相似性(分别为99.7%~99.9%,98.1%~99.7%);该方法可在27h内完成,其灵敏度为2.4~16 CFU/mL;四川省雅安市鸡肉、猪肉、牛肉和牛奶样品中的Cj阳性率分别为38.0%(19/50)、28.3%(15/53)、17.1%(6/35)和8.6%(4/46).

  2. International Clostridium difficile animal strain collection and large diversity of animal associated strains

    DEFF Research Database (Denmark)

    Janezic, Sandra; Zidaric, Valerija; Pardon, Bart

    2014-01-01

    Background: Clostridium difficile is an important cause of intestinal infections in some animal species and animals might be a reservoir for community associated human infections. Here we describe a collection of animal associated C. difficile strains from 12 countries based on inclusion criteria...... of one strain (PCR ribotype) per animal species per laboratory. Results: Altogether 112 isolates were collected and distributed into 38 PCR ribotypes with agarose based approach and 50 PCR ribotypes with sequencer based approach. Four PCR ribotypes were most prevalent in terms of number of isolates...... animal associated C. difficile strains worldwide. The widespread dissemination of toxigenic C. difficile and the considerable overlap in strain distribution between species furthers concerns about interspecies, including zoonotic, transmission of this critically important pathogen....

  3. Kindergarten Animation

    Science.gov (United States)

    Hinshaw, Craig

    2012-01-01

    Animation is one of the last lessons that come to mind when thinking of kindergarten art. The necessary understanding of sequencing, attention to small, often detailed drawings, and the use of technology all seem more suitable to upper elementary. With today's emphasis on condensing and integrating curriculum, consider developing animation lessons…

  4. Animal Detectives

    Science.gov (United States)

    Mulvey, Bridget; Warnock, Carly

    2015-01-01

    During a two-week inquiry-based 5E learning cycle unit, children made observations and inferences to guide their explorations of animal traits and habitats (Bybee 2014). The children became "animal detectives" by studying a live-feed webcam and digital images of wolves in their natural habitat, reading books and online sources about…

  5. Animal ethics

    DEFF Research Database (Denmark)

    Palmer, Clare; Sandøe, Peter

    2011-01-01

    This chapter describes and discusses different views concerning our duties towards animals. First, we explain why it is necessary to engage in thinking about animal ethics and why it is not enough to rely on feelings alone. Secondly, we present and discuss five different kinds of views about...

  6. Apta-PCR.

    Science.gov (United States)

    Pinto, Alessandro; Polo, Pedro Nadal; Rubio, Miriam Jauest; Svobodova, Marketa; Lerga, Teresa Mairal; O'Sullivan, Ciara K

    2016-01-01

    Real-time Apta-PCR is a methodology that can be used for a wide variety of applications ranging from food quality control to clinical diagnostics. This method takes advantage of the combination of the sensitivity of nucleic acid amplification with the selectivity of aptamers. Ultra-low detection of target analyte can potentially be achieved, or, improved detection limits can be achieved with aptamers of low-medium affinity. Herein, we describe a generic methodology coined real-time Apta-PCR, using a model target (β-conglutin) and a competitive format, which can be adapted for the detection of any target which an aptamer has been selected for.

  7. Animal experimentation.

    Science.gov (United States)

    Kolar, Roman

    2006-01-01

    Millions of animals are used every year in often times extremely painful and distressing scientific procedures. Legislation of animal experimentation in modern societies is based on the supposition that this is ethically acceptable when certain more or less defined formal (e.g. logistical, technical) demands and ethical principles are met. The main parameters in this context correspond to the "3Rs" concept as defined by Russel and Burch in 1959, i.e. that all efforts to replace, reduce and refine experiments must be undertaken. The licensing of animal experiments normally requires an ethical evaluation process, often times undertaken by ethics committees. The serious problems in putting this idea into practice include inter alia unclear conditions and standards for ethical decisions, insufficient management of experiments undertaken for specific (e.g. regulatory) purposes, and conflicts of interest of ethics committees' members. There is an ongoing societal debate about ethical issues of animal use in science. Existing EU legislation on animal experimentation for cosmetics testing is an example of both the public will for setting clear limits to animal experiments and the need to further critically examine other fields and aspects of animal experimentation.

  8. Wild Animals

    Institute of Scientific and Technical Information of China (English)

    宁静

    2005-01-01

    Many of us think that all wild animals are dangerous. In fact, very few of them will eat a man if he leaves them alone. If you meet a tiger, I'm sure you will run away, but even a tiger doesn't like meeting a man if it isn't hungry. Tigers only kill and eat man when they are too old to catch their food, such as sheep and other small animals. Some animals get frightened when they only smell a man. Some of themst and and look at a man for a short time before they run away.

  9. Animal Bites

    Science.gov (United States)

    ... to 15 to 20 of every 100 following dog or human bites. Treatment If your child is bleeding from ... dangerous than those from tame, immunized (against rabies) dogs and cats. The health of the animal also is important, so if ...

  10. Desarrollo de un sistema de videoconferencia mediante WebRTC

    OpenAIRE

    Ibarrola Lerga, Josu

    2016-01-01

    El objetivo de este proyecto fin de carrera consiste en el desarrollo de un sistema de videoconferencia mediante WebRTC. Para realizar dicho desarrollo crearemos una página mediante el lenguaje de programación HTML5 y la tecnología que nos brinda WebRTC, todo ello dentro del marco de trabajo de Bootstrap, consiguiendo así una página web apta para cualquier dispositivo de visualización que se utilice. Debido a que se trata de una tecnología nueva que el autor desconoce, para pod...

  11. 检测动物产品中水貂阿留申病毒和犬细小病毒复合PCR方法的建立和应用%Establishment and Application of Multi-PCR Detective Methods for the Detection of Aleutian Mink Disease Virus and Canine Parvovirus in Animal Products

    Institute of Scientific and Technical Information of China (English)

    杜雄伟; 李叶; 胡传伟

    2012-01-01

      水貂阿留申病(AD)和犬细小病毒病(CP)是危害经济动物健康的重要疫病,可造成巨大的经济损失。本研究建立了水貂阿留申病毒(ADV)、犬细小病毒(CPV)复合PCR检测方法,并对临床样品进行了大量检测,结果表明本方法特异、敏感、简便、快速,非常适宜临床样品的大量筛选检测,对经济动物疫情的快速诊断与控制、加强经济动物及其产品进出境的检验检疫工作,御疫情于国门之外有重要的意义。%  Aleutian mink disease and canine parvovirus disease are two important diseases causing great economic loss to fur animal industry. In the study,a multi-PCR assay was developed for detection of Aleutian mink disease and canine parvovirus and the assay was applied in testing a lot of clinical samples. The results showed that the assay was specific,sensitive and rapid,and suitable for screening test.

  12. Evaluation of PCR and multiplex PCR in relation to nested PCR for diagnosing Theileria equi

    Directory of Open Access Journals (Sweden)

    Danielle C. Leal

    2011-07-01

    Full Text Available Conventional PCR (PCRTeq for diagnosing Theileria equi and multiplex PCR (M/PCRTeq-Bc for diagnosing T. equi and Babesia caballi were comparatively evaluated with nested PCR (N/PCR-Teq for diagnosing equine piroplasmosis. In DNA sensitivity determinations, in multiple dilutions of equine blood that had tested positive for T. equi, PCR-Teq and N/PCR-Teq detected hemoparasite DNA in the larger dilutions (1:128, but did not differ significantly from the M/PCRTeq-Bc (1:64. In analyses on equine serum tested by ELISA, there was high agreement between this serological test and PCR-Teq (k = 0.780 and moderate agreement with N/PCR-Teq (k = 0.562 and M/PCRTeq-Bc (k = 0.488. PCR-Teq found a higher frequency of T. equi both in extensively and intensively reared horses, but this was not significant in relation to N/PCR-Teq (P>0.05, and both PCRs indicated that there was an endemic situation regarding T. equi in the population of horses of this sample. PCR-Teq was only significantly different from M/PCR-Teq-Bc (P<0.05. PCR-Teq presented high sensitivity and specificity, comparable to N/PCR-Teq, but with the advantage of higher speed in obtaining results and lower costs and risks of laboratory contamination. This accredits PCR-Teq for epidemiological studies and for determinations on affected horses.

  13. PCR in forensic genetics

    DEFF Research Database (Denmark)

    Morling, Niels

    2009-01-01

    Since the introduction in the mid-1980s of analyses of minisatellites for DNA analyses, a revolution has taken place in forensic genetics. The subsequent invention of the PCR made it possible to develop forensic genetics tools that allow both very informative routine investigations and still more...... and more advanced, special investigations in cases concerning crime, paternity, relationship, disaster victim identification etc. The present review gives an update on the use of DNA investigations in forensic genetics....

  14. Animated symbols

    DEFF Research Database (Denmark)

    Frølunde, Lisbeth

    2008-01-01

    This paper is based on data about animation film production by 18-year-old students in a Danish upper secondary school. The optic is the on-going potential for learning and development of reflection. The purpose is to clarify what might support young people's reflection on media. I propose...... an analytic working model called Animated Symbols concerning critical reflection in a dialogic learning process. The model shows dialogue as interactions that involve two types of transformation: inner ‘learning processes' and outer signs and symbols. The classroom-based research study is part of a Ph...

  15. Comparison of Droplet Digital PCR and qPCR for the Quantification of Shiga Toxin-Producing Escherichia coli in Bovine Feces.

    Science.gov (United States)

    Verhaegen, Bavo; De Reu, Koen; De Zutter, Lieven; Verstraete, Karen; Heyndrickx, Marc; Van Coillie, Els

    2016-05-18

    Cattle are considered to be the main reservoir for Shiga toxin-producing Escherichia coli (STEC) and are often the direct or indirect source of STEC outbreaks in humans. Accurate measurement of the concentration of shed STEC in cattle feces could be a key answer to questions concerning transmission of STEC, contamination sources and efficiency of treatments at farm level. Infected animals can be identified and the contamination level quantified by real-time quantitative PCR (qPCR), which has its specific limitations. Droplet digital PCR (ddPCR) has been proposed as a method to overcome many of the drawbacks of qPCR. This end-point amplification PCR is capable of absolute quantification independent from any reference material and is less prone to PCR inhibition than qPCR. In this study, the qPCR-based protocol described by Verstraete et al. (2014) for Shiga toxin genes stx1 and stx2 and the intimin gene eae quantification was optimized for ddPCR analysis. The properties of ddPCR and qPCR using two different mastermixes (EMM: TaqMan(®) Environmental Master Mix 2.0; UMM: TaqMan(®) Universal PCR Master Mix) were evaluated, using standard curves and both artificial and natural contaminated cattle fecal samples. In addition, the susceptibility of these assays to PCR-inhibitors was investigated. Evaluation of the standard curves and both artificial and natural contaminated cattle fecal samples suggested a very good agreement between qPCR using EMM and ddPCR. Furthermore, similar sensitivities and no PCR inhibition were recorded for both assays. On the other hand, qPCR using UMM was clearly prone to PCR inhibition. In conclusion, the ddPCR technique shows potential for the accurate absolute quantification of STEC on the farms, without relying on standardized reference material.

  16. Comparison of Droplet Digital PCR and qPCR for the Quantification of Shiga Toxin-Producing Escherichia coli in Bovine Feces

    Directory of Open Access Journals (Sweden)

    Bavo Verhaegen

    2016-05-01

    Full Text Available Cattle are considered to be the main reservoir for Shiga toxin-producing Escherichia coli (STEC and are often the direct or indirect source of STEC outbreaks in humans. Accurate measurement of the concentration of shed STEC in cattle feces could be a key answer to questions concerning transmission of STEC, contamination sources and efficiency of treatments at farm level. Infected animals can be identified and the contamination level quantified by real-time quantitative PCR (qPCR, which has its specific limitations. Droplet digital PCR (ddPCR has been proposed as a method to overcome many of the drawbacks of qPCR. This end-point amplification PCR is capable of absolute quantification independent from any reference material and is less prone to PCR inhibition than qPCR. In this study, the qPCR-based protocol described by Verstraete et al. (2014 for Shiga toxin genes stx1 and stx2 and the intimin gene eae quantification was optimized for ddPCR analysis. The properties of ddPCR and qPCR using two different mastermixes (EMM: TaqMan® Environmental Master Mix 2.0; UMM: TaqMan® Universal PCR Master Mix were evaluated, using standard curves and both artificial and natural contaminated cattle fecal samples. In addition, the susceptibility of these assays to PCR-inhibitors was investigated. Evaluation of the standard curves and both artificial and natural contaminated cattle fecal samples suggested a very good agreement between qPCR using EMM and ddPCR. Furthermore, similar sensitivities and no PCR inhibition were recorded for both assays. On the other hand, qPCR using UMM was clearly prone to PCR inhibition. In conclusion, the ddPCR technique shows potential for the accurate absolute quantification of STEC on the farms, without relying on standardized reference material.

  17. Animal house

    OpenAIRE

    Turka, Laurence A.

    2008-01-01

    While the JCI was originally conceived as a journal that would integrate various scientific approaches to the examination of human physiology and pathophysiology, we now find many of its pages filled with animal models of human disease. Is this a good thing?

  18. Animated Symbols

    DEFF Research Database (Denmark)

    Frolunde, Lisbeth

    ' processer af fem udvalgte elever er gennemgået i forhold til tre opdelinger: filmskabere, filmskabelse processen og film. Den teoretiske tilgang er pragmatisme, social semiotik og diskursanalyse. Modellen "Animating Symbols" er udviklet og diskuteret som forsøg på at forstå reflektion og design som en slags...

  19. A PCR amplification method without DNA extraction.

    Science.gov (United States)

    Li, Hongwei; Xu, Haiyue; Zhao, Chunjiang; Sulaiman, Yiming; Wu, Changxin

    2011-02-01

    To develop a simple and inexpensive method for direct PCR amplification of animal DNA from tissues, we optimized different components and their concentration in lysis buffer systems. Finally, we acquired the optimized buffer system composed of 10 mmol tris(hydroxymethyl)aminomethane (Tris)-Cl (pH 8.0), 2 mmol ethylene diamine tetraacetic (EDTA) (pH 8.0), 0.2 mol NaCl and 200 μg/mL Proteinase K. Interestingly, the optimized buffer is also very effective when working with common human sample types, including blood, buccal cells and hair. The direct PCR method requires fewer reagents (Tris-Cl, EDTA, Protease K and NaCl) and less incubation time (only 35 min). The cost of treating every sample is less than $0.02, and all steps can be completed on a thermal cycler in a 96-well format. So, the proposed method will significantly improve high-throughput PCR-based molecular assays in animal systems and in common human sample types.

  20. Biotecnologia animal

    Directory of Open Access Journals (Sweden)

    Luiz Lehmann Coutinho

    2010-01-01

    Full Text Available A biotecnologia animal tem fornecido novas ferramentas para os programas de melhoramento e, dessa forma, contribuído para melhorar a eficiência da produção dos produtos de origem animal. No entanto, os avanços têm sido mais lentos do que antecipados, especialmente em razão da dificuldade na identificação dos genes responsáveis pelas características fenotípicas de interesse zootécnico. Três estratégias principais têm sido utilizadas para identificar esses genes - mapeamento de QTL, genes candidatos e sequenciamento de DNA e mRNA - e cada uma tem suas vantagens e limitações. O mapeamento de QTL permite determinar as regiões genômicas que contêm genes, mas o intervalo de confiança do QTL pode ser grande e conter muitos genes. A estratégia de genes candidatos é limitada por causa do conhecimento ainda restrito das funções de todos os genes. Os sequenciamentos de genomas e de sequências expressas podem auxiliar na identificação da posição de genes e de vias metabólicas associadas à característica de interesse. A integração dessas estratégias por meio do desenvolvimento de programas de bioinformática permitirá a identificação de novos genes de interesse zootécnico. Assim, os programas de melhoramento genético se beneficiarão pela inclusão da informação obtida diretamente do DNA na avaliação do mérito genético dos plantéis disponíveis.Animal biotechnology is providing new tools for animal breeding and genetics and thus contributing to advances in production efficiency and quality of animal products. However, the progress is slower than anticipated, mainly because of the difficulty involved in identifying genes that control phenotypic characteristics of importance to the animal industry. Three main strategies: QTL mapping, candidate genes and DNA and mRNA sequencing have been used to identify genes of economic interest to animal breeding and each has advantages and disadvantages. QTL mapping allows

  1. Animal Locomotion

    CERN Document Server

    Taylor, Graham K; Tropea, Cameron

    2010-01-01

    This book provides a wide-ranging snapshot of the state-of-the-art in experimental research on the physics of swimming and flying animals. The resulting picture reflects not only upon the questions that are of interest in current pure and applied research, but also upon the experimental techniques that are available to answer them. Doubtless, many new questions will present themselves as the scope and performance of our experimental toolbox develops over the coming years.

  2. PCR, exit stage left ...

    CERN Multimedia

    2004-01-01

    The Prevessin Control Room during LEP's start up in 1989. The Prévessin Control Room (PCR) was recently engulfed in a wave of nostalgia. The PCR, scene of some of the greatest moments in CERN's history, is being dismantled to prepare for a complete overhaul. In February 2006, a new combined control centre for all the accelerators will open its doors on the same site, together with a new building currently under construction (see Bulletin issue 27/2004 of 28 June 2004). This marks the end of an important chapter in CERN's history. The Prévessin Control Room saw its first momentous event 28 years ago when the 400 GeV beam for the SPS was commissioned in the presence of Project Leader John Adams. It was also here that the first proton-antiproton collisions were observed, in 1981. Eight years later, in 1989, operators and directors alike jumped for joy at the announcement of the first electron-positron collisions at the start up of LEP, the biggest accelerator in the world. Today the 80 terminals and PCs have b...

  3. Comparison of conventional PCR, quantitative PCR, bacteriological culture and the Warthin Starry technique to detect Leptospira spp. in kidney and liver samples from naturally infected sheep from Brazil.

    Science.gov (United States)

    Fornazari, Felipe; da Silva, Rodrigo Costa; Richini-Pereira, Virginia Bodelão; Beserra, Hugo Enrique Orsini; Luvizotto, Maria Cecília Rui; Langoni, Helio

    2012-09-01

    Leptospirosis is an infectious disease of worldwide importance. The development of diagnostic techniques allows sick animals to be identified, reservoirs to be eliminated and the disease prevented and controlled. The present study aimed to compare different techniques for diagnosing leptospirosis in sheep. Samples of kidney, liver and blood were collected from 465 animals that originated from a slaughterhouse. The sera were analyzed by the Microscopic Agglutination Test (MAT), and kidney and liver samples of seropositive animals were analyzed using four techniques: bacteriological culture, the Warthin Starry (WS) technique, conventional PCR (cPCR), and quantitative PCR (qPCR). With the MAT, 21 animals were positive (4.5%) to serovars Hardjo (n=12), Hebdomadis (n=5), Sentot (n=2), Wolfii (n=1) and Shermani (n=1). Titers were 100 (n=10), 200 (n=2), 400 (n=6) and 1600 (n=3). No animal was positive by bacteriological culture; four animals were positive by the WS technique in kidney samples; six animals were positive by cPCR in kidney samples; and 11 animals were positive by qPCR, eight of which in kidney samples and three in liver. The bacterial quantification revealed a median of 4.3 bacteria/μL in liver samples and 36.6 bacteria/μL in kidney samples. qPCR presented the highest sensitivity among the techniques, followed by cPCR, the WS technique and bacteriological culture. These results indicate that sheep can carry leptospires of the Sejroe serogroup, and demonstrate the efficiency of quantitative PCR to detect Leptospira spp. in tissue samples.

  4. Animal Testing

    Science.gov (United States)

    Moretto, Johnny; Chauffert, Bruno; Bouyer, Florence

    The development of a new anticancer drug is a long, complex and multistep process which is supervised by regulatory authorities from the different countries all around the world [1]. Application of a new drug for admission to the market is supported by preclinical and clinical data, both including the determination of pharmacodynamics, toxicity, antitumour activity, therapeutic index, etc. As preclinical studies are associated with high cost, optimization of animal experiments is crucial for the overall development of a new anticancer agent. Moreover, in vivo efficacy studies remain a determinant panel for advancement of agents to human trials and thus, require cautious design and interpretation from experimental and ethical point of views.

  5. Animation & Neurocinematics*

    DEFF Research Database (Denmark)

    Carpe Pérez, Inmaculada Concepción

    2016-01-01

    We love movies because we like to jump from our “reality” to live a dream, a parallel universe that inspires us. We long for adventure, excitement and answers to quests… That’s the magic of cinema; it makes you believe what you see and over all, FEEL it. As Antonio Damasio said-“ we´re feeling...... machines that think”-(Damasio, A. Descartes error). Such feelings come from the interpretation of the emotions in our bodies. Emotions are our universal language, the motivation of living, the key to what makes a movie successful and truly an art piece that you will remember because moves you. Animation...

  6. PCR em tempo real para diagnóstico da leucose enzoótica bovina Enzootic bovine leukosis real time PCR

    Directory of Open Access Journals (Sweden)

    Natanael Lamas Dias

    2012-08-01

    Full Text Available O objetivo deste trabalho foi realizar a validação de uma reação em cadeia da polimerase em tempo real com o sistema Plexor® (qPCR para o diagnóstico da Leucose Enzoótica Bovina (LEB, por meio da comparação com testes de diagnóstico recomendados pela Organização Mundial de Saúde Animal (OIE. A qPCR foi comparada com duas outras técnicas: a PCR nested (nPCR e a imunodifusão em gel de ágar (IDGA. Das 82 amostras analisadas pela qPCR e nPCR, 79 apresentaram resultados concordantes, sendo a concordância, classificada pelo Índice Kappa, como alta. Entre as PCRs e a IDGA, o número de resultados concordantes foi de 71 e 69, respectivamente, para qPCR e nPCR, sendo a concordância classificada como considerável. A qPCR apresentou altos valores de sensibilidade e especificidade. Os valores preditivos da qPCR observados demonstraram a alta capacidade de classificação dos casos positivos e negativos. A qPCR não foi capaz de detectar três amostras positivas e tem custo ligeiramente superior que a nPCR. Entretanto, a qPCR é uma técnica mais rápida, menos susceptível a contaminações, tem alta sensibilidade, não utiliza e não gera resíduos carcinogênicos. Concluímos que a qPCR pode substituir a nPCR recomendada pela OIE no diagnóstico de rotina em áreas em que a LEB é endêmica, como no Brasil.The goal of this research was to validate a Plexor® real time Polymerase Chain Reaction (qPCR for Enzootic Bovine Leukosis (EBL diagnosis by comparison with methods recommend by the World Animal Health Organization (OIE. The qPCR was compared with two other techniques: the nested PCR (nPCR and to the agar gel immunodiffusion (AGID. Of 82 qPCR and nPCR analysed samples, 79 presented concordant results, being the concordance classified by Kappa Index as high. Between the PCRs and AGID, the number of concordant results was 71 and 69, out of 82, to qPCR and nPCR, respectively, being the concordance classified as considerable, in both

  7. Detección molecular de las translocaciones más comunes en Leucemia aguda mediante RT-PCR

    Directory of Open Access Journals (Sweden)

    Guevara G.

    2001-06-01

    Full Text Available Evaluar la incidencia de las translocaciones t(4;11, t(1;19, t(9;22 y t(12;21 en leucemia linfoide aguda (LLA y t(15;17, t(8;21 e Inv.(16 en leucemia mieloide aguda (LMA. Correlacionar los resultados obtenidos con el diagnóstico morfológico y citogenético.

  8. Detección del virus de la leucosis bovina en ganado criollo colombiano mediante PCR-anidado

    Directory of Open Access Journals (Sweden)

    Giovambattista Guillermo

    2011-12-01

    Full Text Available

    Se evaluó la presencia del virus de la leucosis bovina (VLB en 360 muestras de ADN de oc%o razas bovinas criollas: Blanco (reinegro  (B(N,  -asanare/o (-AS,  -oste/o con -uernos (---, -%ino Santandereano (-%S, -a4uete/o (-8T, Hartón del Valle (HV, Romosinuano (RS y San Martinero (SM, dos Razas SintBticas -olombianas: Lucerna (LD-  y VelEs4uez (VGL y dos razas HorEneas: Bra%- mEn (B y Holstein (HJ Para la detección del proKvirus se amplificó una región del gen env viral, me- diante P-R  anidadaJ La presencia del VLB Hue mayor en la raza HV seguido por -%S (M3J3O y 60O respectivamente, VGL y LD-  tuvieron el mismo porcentae (Q0O, en -AS,  --- y -8T la presencia del virus fue de 26.7%, 23.3% y 16.7% respectivamente; no se encontró el virus en BON, SM y RS. En las razas foráneas la presencia fue de 83.3% para H y 6.7% para B. Se encontró dependencia altamente significativa entre la presencia del VLB y la raza, el sexo y región de origen de la muestraJ Gl prome- dio de presencia en las razas criollas Hue menor 4ue en las HorEneas, menor en los mac%os 4ue en las síembras y en la región norte 4ue en el suroccidente y el centro del pais

  9. Bioethical Problems: Animal Welfare, Animal Rights.

    Science.gov (United States)

    March, B. E.

    1984-01-01

    Discusses various bioethical issues and problems related to animal welfare and animal rights. Areas examined include: Aristotelian views; animal welfare legislation; Darwin and evolutionary theory; animal and human behavior; and vegetarianism. A 14-point universal declaration of the rights of animals is included. (JN)

  10. COMPARISON OF 16S rRNA-PCR-RFLP, LipL32-PCR AND OmpL1-PCR METHODS IN THE DIAGNOSIS OF LEPTOSPIROSIS

    Directory of Open Access Journals (Sweden)

    Tülin GÜVEN GÖKMEN

    Full Text Available SUMMARY Leptospirosis is still one of the most important health problems in developing countries located in humid tropical and subtropical regions. Human infections are generally caused by exposure to water, soil or food contaminated with the urine of infected wild and domestic animals such as rodents and dogs. The clinical course of leptospirosis is variable and may be difficult to distinguish from many other infectious diseases. The dark-field microscopy (DFM, serology and nucleic acid amplification techniques are used to diagnose leptospirosis, however, a distinctive standard reference method is still lacking. Therefore, in this study, we aimed to determine the presence of Leptospira spp., to differentiate the pathogenic L. interrogans and the non-pathogenic L. biflexa, and also to determine the sensitivity and specificity values of molecular methods as an alternative to conventional ones. A total of 133 serum samples, from 47 humans and 86 cattle were evaluated by two conventional tests: the Microagglutination Test (MAT and the DFM, as well as three molecular methods, the 16S rRNA-PCR followed by Restriction Fragment Lenght Polymorphism (RFLP of the amplification products 16S rRNA-PCR-RFLP, LipL32-PCR and OmpL1-PCR. In this study, for L. interrogans, the specificity and sensitivity rates of the 16S rRNA-PCR and the LipL32-PCR were considered similar (100% versus 98.25% and 100% versus 98.68%, respectively. The OmpL1-PCR was able to classify L. interrogans into two intergroups, but this PCR was less sensitive (87.01% than the other two PCR methods. The 16S rRNA-PCR-RFLP could detect L. biflexa DNA, but LipL32-PCR and OmpL1-PCR could not. The 16S rRNA-PCR-RFLP provided an early and accurate diagnosis and was able to distinguish pathogenic and non-pathogenic Leptospira species, hence it may be used as an alternative method to the conventional gold standard techniques for the rapid disgnosis of leptospirosis.

  11. COMPARISON OF 16S rRNA-PCR-RFLP, LipL32-PCR AND OmpL1-PCR METHODS IN THE DIAGNOSIS OF LEPTOSPIROSIS

    Science.gov (United States)

    GÖKMEN, Tülin GÜVEN; SOYAL, Ayben; KALAYCI, Yıldız; ÖNLEN, Cansu; KÖKSAL, Fatih

    2016-01-01

    SUMMARY Leptospirosis is still one of the most important health problems in developing countries located in humid tropical and subtropical regions. Human infections are generally caused by exposure to water, soil or food contaminated with the urine of infected wild and domestic animals such as rodents and dogs. The clinical course of leptospirosis is variable and may be difficult to distinguish from many other infectious diseases. The dark-field microscopy (DFM), serology and nucleic acid amplification techniques are used to diagnose leptospirosis, however, a distinctive standard reference method is still lacking. Therefore, in this study, we aimed to determine the presence of Leptospira spp., to differentiate the pathogenic L. interrogans and the non-pathogenic L. biflexa, and also to determine the sensitivity and specificity values of molecular methods as an alternative to conventional ones. A total of 133 serum samples, from 47 humans and 86 cattle were evaluated by two conventional tests: the Microagglutination Test (MAT) and the DFM, as well as three molecular methods, the 16S rRNA-PCR followed by Restriction Fragment Lenght Polymorphism (RFLP) of the amplification products 16S rRNA-PCR-RFLP, LipL32-PCR and OmpL1-PCR. In this study, for L. interrogans, the specificity and sensitivity rates of the 16S rRNA-PCR and the LipL32-PCR were considered similar (100% versus 98.25% and 100% versus 98.68%, respectively). The OmpL1-PCR was able to classify L. interrogans into two intergroups, but this PCR was less sensitive (87.01%) than the other two PCR methods. The 16S rRNA-PCR-RFLP could detect L. biflexa DNA, but LipL32-PCR and OmpL1-PCR could not. The 16S rRNA-PCR-RFLP provided an early and accurate diagnosis and was able to distinguish pathogenic and non-pathogenic Leptospira species, hence it may be used as an alternative method to the conventional gold standard techniques for the rapid disgnosis of leptospirosis. PMID:27680169

  12. Animating Brains

    Science.gov (United States)

    Borck, Cornelius

    2016-01-01

    A recent paper famously accused the rising field of social neuroscience of using faulty statistics under the catchy title ‘Voodoo Correlations in Social Neuroscience’. This Special Issue invites us to take this claim as the starting point for a cross-cultural analysis: in which meaningful ways can recent research in the burgeoning field of functional imaging be described as, contrasted with, or simply compared to animistic practices? And what light does such a reading shed on the dynamics and effectiveness of a century of brain research into higher mental functions? Reviewing the heated debate from 2009 around recent trends in neuroimaging as a possible candidate for current instances of ‘soul catching’, the paper will then compare these forms of primarily image-based brain research with older regimes, revolving around the deciphering of the brain’s electrical activity. How has the move from a decoding paradigm to a representational regime affected the conceptualisation of self, psyche, mind and soul (if there still is such an entity)? And in what ways does modern technoscience provide new tools for animating brains? PMID:27292322

  13. Animal welfare: an animal science approach.

    Science.gov (United States)

    Koknaroglu, H; Akunal, T

    2013-12-01

    Increasing world population and demand for animal-derived protein puts pressure on animal production to meet this demand. For this purpose animal breeding efforts were conducted to obtain the maximum yield that the genetic makeup of the animals permits. Under the influence of economics which is the driving force behind animal production, animal farming became more concentrated and controlled which resulted in rearing animals under confinement. Since more attention was given on economics and yield per animal, animal welfare and behavior were neglected. Animal welfare which can be defined as providing environmental conditions in which animals can display all their natural behaviors in nature started gaining importance in recent years. This does not necessarily mean that animals provided with good management practices would have better welfare conditions as some animals may be distressed even though they are in good environmental conditions. Consumers are willing to pay more for welfare-friendly products (e.g.: free range vs caged egg) and this will change the animal production practices in the future. Thus animal scientists will have to adapt themselves for the changing animal welfare rules and regulations that differ for farm animal species and countries. In this review paper, animal welfare is discussed from an animal science standpoint.

  14. Quantitative PCR for glucose transporter and tristetraprolin family gene expression in cultured mouse adipocytes and macrophages.

    Science.gov (United States)

    Cao, Heping; Cao, Fangping; Roussel, Anne-Marie; Anderson, Richard A

    2013-12-01

    Quantitative real-time PCR (qPCR) such as TaqMan and SYBR Green qPCR are widely used for gene expression analysis. The drawbacks of SYBR Green assay are that the dye binds to any double-stranded DNA which can generate false-positive signals and that the length of the amplicon affects the intensity of the amplification. Previous results demonstrate that TaqMan assay is more sensitive but generates lower calculated expression levels than SYBR Green assay in quantifying seven mRNAs in tung tree tissues. The objective of this study is to expand the analysis using animal cells. We compared both qPCR assays for quantifying 24 mRNAs including those coding for glucose transporter (Glut) and mRNA-binding protein tristetraprolin (TTP) in mouse 3T3-L1 adipocytes and RAW264.7 macrophages. The results showed that SYBR Green and TaqMan qPCR were reliable for quantitative gene expression in animal cells. This result was supported by validation analysis of Glut and TTP family gene expression. However, SYBR Green qPCR overestimated the expression levels in most of the genes tested. Finally, both qPCR instruments (Bio-Rad's CFX96 real-time system and Applied Biosystems' Prism 7700 real-time PCR instrument) generated similar gene expression profiles in the mouse cells. These results support the conclusion that both qPCR assays (TaqMan and SYBR Green qPCR) and both qPCR instruments (Bio-Rad's CFX96 real-time system and Applied Biosystems' Prism 7700 real-time PCR instrument) are reliable for quantitative gene expression analyses in animal cells but SYBR Green qPCR generally overestimates gene expression levels than TaqMan qPCR.

  15. Análisis taxonómico y funcional del microbioma humano mediante aproximaciones clásicas, moleculares y metagenómicas

    OpenAIRE

    Cabrera Rubio, Raúl

    2014-01-01

    La presente tesis muestra distintas aproximaciones para el estudio del microbioma humano. Éstas han ido desde la secuenciación masiva de productos de PCR, la pirosecuenciación directa del ADN ambiental, la elaboración de librerías de fósmidos y por último el aislamiento de especies presentes en el microbioma mediante sembrado de la muestra. Todas estas técnicas tienen sus ventajas y desventajas, pero todas ellas son complementarias para el estudio de un determinado microbioma. Además la elabo...

  16. 动物性食品中沙门菌实时荧光定量 PCR快速检测方法的建立%Development of Real-time PCR for Rapid Detection of Salmonella spp in Animal-origined Food

    Institute of Scientific and Technical Information of China (English)

    李丹丹; 徐义刚; 王绥家; 高慎阳; 马广鹏

    2014-01-01

    A real-time PCR was developed for the rapid detection of Salmonella spp based on the primers and probes which were designed for the conservative domain of fimY gene of Salmonella spp.The sensitiv-ity of the real-time PCR was 5.2 cfu/mL.56 from 781 samples of meat,eggs,milk aquatic products were positive in real-time PCR assay,which was in accordance with the testing result according to GB/T4789.4-2008.The results showed that a real-time PCR assay was sensitive,rapid and simple.%根据沙门菌属高度保守的 fimY 基因序列设计探针和引物,通过优化反应条件,建立检测动物性食品中沙门菌实时荧光定量 PCR 方法,应用于动物性食品中沙门菌的快速检验。建立的荧光 PCR 方法灵敏度约为5.2 cfu/mL,经对781份肉类、蛋、奶及水产品进行检测,共检出56份阳性样本,与国标(GB/T4789.4---2008)方法的检测结果一致。建立的荧光 PCR 方法操作简便、特异性强、灵敏度高,具有良好的实用性。

  17. en sistemas modelo de gelatina mediante SPME-DED

    Directory of Open Access Journals (Sweden)

    J. Ruiz Carrascal

    2005-01-01

    Full Text Available Se llevó a cabo la determinación de 16 hidrocarburos aromáticos policíclicos y 9 nitrosaminas en sistemas modelo de gelatina (20% w/v mediante microextración en fase sólida acoplada a un dispositivo de extracción directa y posterior análisis mediante cromatografía-gaseosa-espectrometría de masas. La extracción se llevo a cabo a 25ºC. Se extrajeron la totalidad de las nitrosaminas y 9 de los 16 hidrocarburos aromáticos policíclicos. Se evaluaron la reproducibilidad, linealidad de respuesta y límite de detección de 3 tipos de fibras estacionarias para cada tipo de compuesto. Se seleccionaron las fibras de polidimetilsiloxano 100μm y carboxen/polidimetilsiloxano 85μm, para los hidrocarburos aromáticos policiclicos y nitrosaminas, respectivamente. La microextración en fase sólida acoplada a un dispositivo de extracción directa surge como una técnica interesante para la monitorización preliminar de la presencia de estos compuestos tóxicos en alimentos sólidos, sin necesidad de toma de muestras y sin deteriorar el producto

  18. Trypanosoma spp. in Swedish game animals.

    Science.gov (United States)

    Neumüller, Magnus; Nilsson, Kenneth; Påhlson, Carl

    2012-01-01

    Serum and blood samples from 36 game animals, shot during the hunting seasons 2007-2009, were collected and analyzed for the presence of Trypanosoma spp. by three methods: isolation, polymerase chain reaction (PCR), and serology. Only fissiped animals were included, four different ruminants and wild boar. Trypanosomes could be isolated from two of the animals, and eight had detectable parasite DNA. Seven animals had high titers of anti-trypanosoma IgG antibodies. The two isolated strains, one from roe dear and one from European elk, were determined to Trypanosoma theileri by partial DNA sequencing of the 18S ribosomal gene. In the seven boars, no Trypanosoma were detected, but four out of seven strongly positive serological samples came from this group. This is the first study in Scandinavia on the presence of Trypanosoma in game animals. The results indicate that trypanosomiasis is frequently occurring among Swedish game animals.

  19. Evaluation of reverse transcription-PCR protocols based on the fusion gene for diagnosis of bovine respiratory syncytial virus infections

    OpenAIRE

    Selim A.; Gaede W.

    2013-01-01

    Bovine respiratory syncytial virus (BRSV) is a pneumovirus in the family paramyxoviridae, is an important cause of acute respiratory disease in postweaning calves and feedlot cattle. The real-time reverse transcriptase PCR protocols were developed to detect BRSV infection in infected animals. The sensitivity of RT-PCR protocols based on fusion gene were evaluated using different Mastermixes such as Qiagen One Step RT-PCR (Qiagen) for conventional RT-PCR, Su...

  20. Digital PCR: A brief history

    OpenAIRE

    2014-01-01

    Digital PCR for quantification of a target of interest has been independently developed several times, being described in 1990 and 1991 using the term “limiting dilution PCR” and in 1999 using the term “digital PCR”. It came into use in the decade following its first development but its use was cut short by the description of real-time PCR in 1996. However digital PCR has now had a renaissance due to the recent development of new instruments and chemistry which have made it a much simpler and...

  1. The wild animal as a research animal

    NARCIS (Netherlands)

    Swart, JAA

    2004-01-01

    Most discussions on animal experimentation refer to domesticated animals and regulations are tailored to this class of animals. However, wild animals are also used for research, e. g., in biological field research that is often directed to fundamental ecological-evolutionary questions or to conserva

  2. Animation of Antimicrobial Resistance

    Medline Plus

    Full Text Available ... Veterinary Home Animal & Veterinary Safety & Health Antimicrobial Resistance Animation of Antimicrobial Resistance Share Tweet Linkedin Pin it ... Veterinary Medicine is cited as the corporate author. Animation Animation of Antimicrobial Resistance (WMV - 19.2MB) 9: ...

  3. Learning Anime Studio

    CERN Document Server

    Troftgruben, Chad

    2014-01-01

    Anime Studio is your complete animation program to help you create 2D movies, cartoons, anime, and cut out animations. You can create your own animated shorts and use Anime Studio to produce cartoon animations for film, video, or streaming over the Web, which can be enjoyed on YouTube, Vimeo, and other popular sites. Anime Studio is great for hobbyists and professionals alike, combining tools for both illustration and animation. With Anime Studio's easy-to-use interface, you will be creating an animated masterpiece in no time. This practical, step-by-step guide will provide you with a structur

  4. Optimized PCR assay for detection of white spot syndrome virus (WSSV).

    Science.gov (United States)

    Nunan, Linda M; Lightner, Donald V

    2011-01-01

    A rapid PCR assay for detection of white spot syndrome virus (WSSV) was developed based on the nested PCR procedure described by Lo et al. (1996) and outlined as the recommended PCR diagnostic assay in the Manual of Diagnostic Tests for Aquatic Animals published by the Office of International Epizootics (OIE, 2009). The optimized procedure incorporated the second step primers used in the nested WSSV PCR. By adjusting the annealing temperature and shortening the cycling times, this modified assay is substantially faster and as sensitive as the recommended OIE protocol. The modified PCR test was compared directly to the two-step nested PCR protocol and a modified nested procedure. The sensitivity of the published assay was determined by template dilutions of semi-purified WSSV virions that had been quantitated using real-time PCR for detection of WSSV. Various isolates were tested using the modified procedure, to ensure that the assay was able to detect WSSV from different geographical locations.

  5. Multiplex SYBR® green-real time PCR (qPCR) assay for the detection and differentiation of Bartonella henselae and Bartonella clarridgeiae in cats.

    Science.gov (United States)

    Staggemeier, Rodrigo; Pilger, Diogo André; Spilki, Fernando Rosado; Cantarelli, Vlademir Vicente

    2014-01-01

    A novel SYBR® green-real time polymerase chain reaction (qPCR) was developed to detect two Bartonella species, B. henselae and B. clarridgeiae, directly from blood samples. The test was used in blood samples obtained from cats living in animal shelters in Southern Brazil. Results were compared with those obtained by conventional PCR targeting Bartonella spp. Among the 47 samples analyzed, eight were positive using the conventional PCR and 12 were positive using qPCR. Importantly, the new qPCR detected the presence of both B. henselae and B. clarridgeiae in two samples. The results show that the qPCR described here may be a reliable tool for the screening and differentiation of two important Bartonella species.

  6. MULTIPLEX SYBR® GREEN-REAL TIME PCR (qPCR ASSAY FOR THE DETECTION AND DIFFERENTIATION OF Bartonella henselae AND Bartonella clarridgeiae IN CATS

    Directory of Open Access Journals (Sweden)

    Rodrigo Staggemeier

    2014-04-01

    Full Text Available A novel SYBR® green-real time polymerase chain reaction (qPCR was developed to detect two Bartonella species, B. henselae and B. clarridgeiae, directly from blood samples. The test was used in blood samples obtained from cats living in animal shelters in Southern Brazil. Results were compared with those obtained by conventional PCR targeting Bartonella spp. Among the 47 samples analyzed, eight were positive using the conventional PCR and 12 were positive using qPCR. Importantly, the new qPCR detected the presence of both B. henselae and B. clarridgeiae in two samples. The results show that the qPCR described here may be a reliable tool for the screening and differentiation of two important Bartonella species.

  7. Detección de Mycoplasma genitalium mediante Reacción en Cadena de la Polimerasa en muestras urogenitales de individuos cubanos sexualmente activos

    Directory of Open Access Journals (Sweden)

    Brian Arturo Mondeja-Rodríguez

    2014-04-01

    Full Text Available El diagnóstico de las infecciones por Mycoplasma genitalium mediante métodos bacteriológicos tradicionales resulta laborioso y poco práctico. Es por ello que los métodos moleculares basados en la amplificación del ADN se utilizan con fines diagnósticos de las infecciones causadas por este microorganismo. En Cuba se han realizado pocos estudios sobre la presencia de M. genitalium en el tracto urogenital. El objetivo de la presente investigación fue detectar M. genitalium en individuos cubanos sexualmente activos mediante la implementación de métodos de PCR simple. Se implementaron dos PCR simples para la detección de fragmentos de 427 pb del gen ARN ribosomal 16S y 281 pb del gen de la adhesina celular MgPa de M. genitalium, que se evaluaron en muestras de exudado endocervical provenientes de 300 mujeres con sintomatología urogenital y muestras de orina de 49 hombres asintomáticos sexualmente activos. Se logró un límite de detección de la PCR del ARNr 16S de aproximadamente 5 copias de genoma por reacción, mientras que para la PCR MgPa se logró la amplificación de solo 50 copias de genoma por reacción. El 3% (10/300 de los exudados endocervicales y el 24,5% (12/49 de las muestras de orina de hombres asintomáticos resultaron positivas mediante ambas PCR. El mayor porcentaje de muestras positivas correspondió a las muestras de orina provenientes de hombres asintomáticos, que resultó superior a lo esperado. El presente trabajo permitirá realizar estudios futuros de caracterización genética y antigénica de las cepas de Mycoplasma genitalium circulantes en Cuba, útiles para conformar un inmunógeno vacunal.

  8. MEDIANTE LA APLICACIÓN DEL ENSAYO FÉNIX.

    Directory of Open Access Journals (Sweden)

    Linna Marcela Neme Ardila

    2013-01-01

    Full Text Available Las mezclas asfálticas son el material más utilizado en la fabricación de pavimentos y los ensayos que permiten caracterizarlas son costosos y demorados. Por esta razón, mediante esta investigación se planteó establecer la viabilidad del uso del ensayo Fénix en mezclas asfálticas colombianas con granulometrías del Instituto de Desarrollo Urbano (IDU y del Instituto Nacional de Vías (INVIAS con diferentes características. El estudio inició con la fabricación de probetas Fénix con diferentes materiales (Agregados, asfaltos, asfaltita, pavimento asfalto reciclado (RAP, cal, cemento y su ejecución a 15 °C, una velocidad de 1 mm/min y la medición de los parámetros del ensayo. De los resultados obtenidos de resistencia a tracción, índice de rigidez a tracción e índice de energía, área elástica y área de fluencia, se estableció que el ensayo Fénix es un procedimiento eficaz, eficiente, económico, rápido y sencillo para determinar las propiedades mecánicas y dinámicas de las mezclas asfálticas estudiadas, especialmente mediante el análisis de los parámetros de las curvas carga-desplazamiento, irrelevantemente de la mezcla asfáltica fabricada.

  9. Importancia del bienestar animal en las unidades de producción animal en México - Importance of animal welfare in units of animal production in México

    OpenAIRE

    2009-01-01

    ResumenEn la actualidad, el bienestar animal (BA), es un tema de vitalimportancia a tomar en cuenta en las Unidades de Producción Animal(UPAS), cuya importancia está relacionado con el trato que el hombrele proporciona a los animales, tanto en la movilización para el manejoen las UPAS y el transporte para el sacrificio, en cualquier parte delmundo. Mediante el uso de conocimientos científicos, relacionadoscon la importancia que tienen el BA para el buen desempeñoreproductivo y productivo de l...

  10. L'adquisició de competències narratives i audiovisuals mitjançant el llibre il·lustrat, la tira còmica i el cinema d'animació Acquiring Narrative and Audiovisual Competences through the Use of Storybooks, Comic Strips and Animated Films La adquisición de competencias narrativas y audiovisuales mediante el libre ilustrado, la tira cómica y el cine de animación

    Directory of Open Access Journals (Sweden)

    Teresa Duran i Armengol

    2008-01-01

    Full Text Available El propòsit d’aquest estudi té un doble objectiu: d’una banda, des del punt de vista històric, assenyalar la confluència existent, a cavall dels segles XIX i XX, d’un magma comú de pràctiques del dibuix que permeteren l’eclosió de certes arts narratives com el llibre il·lustrat, la tira còmica i el cinema d’animació, materials que cent anys després de la seva gènesi encara no han trobat el seu lloc en l’ensenyament i, de l’altra, des del punt de vista educatiu, aportar instruments de reflexió per a l’adquisició de millors competències narratives i audiovisuals a partir d’aquests materials. _____________________________________________ Le but de cette étude est double : d’une part, du point de vue historique, signaler la confluence existant, à cheval sur les XIXe et XXe siècles, dans un magma commun de pratiques du dessin qui permettent l’éclosion de certains arts narratifs tels que le livre illustré, la bande dessinée ou le cinéma d’animation – trois matériels qui, cent ans après leur naissance, n’ont pas encore pas trouvé leur place dans l’enseignement – et, de l’autre, du point de vue éducatif, apporter les instruments de réflexion pour l’acquisition de meilleures compétences narratives et audiovisuelles à partir de ces matériels.This study has two aims. From a historical viewpoint, it explores the confluence of a wide range of drawing practices at the end of the nineteenth century and the beginning of the twentieth, which led to a blossoming of narrative arts such as the storybook, the comic strip and the animated movie. Now, a hundred years since their inception, these genres have still not found their place in the syllabus. From an educational viewpoint, the article offers tools for considering the acquisition of better narrative and audiovisual competences based on the use of these genres.El propósito de este artículo tiene un doble objetivo: por un lado, desde el punto de

  11. A mitleidsethik e os animais ou schopenhauer como precursos da ética animal

    Directory of Open Access Journals (Sweden)

    Jair Barboza

    2010-12-01

    Full Text Available http://dx.doi.org/10.5007/1677-2954.2008v7n2p253Este artifo tem por objetivo mostrar que Schopenhauer, mediante sua Mitleidsethik (ética da compaixão, baseada numa metafísica da Vontade de vida, pode ser visto como um precursor da ética animal.

  12. Importancia del bienestar animal en las unidades de producción animal en México

    OpenAIRE

    2009-01-01

    En la actualidad, el bienestar animal (BA), es un tema de vital importancia a tomar en cuenta en las Unidades de Producción Animal (UPAS), cuya importancia está relacionado con el trato que el hombre le proporciona a los animales, tanto en la movilización para el manejo en las UPAS y el transporte para el sacrificio, en cualquier parte del mundo. Mediante el uso de conocimientos científicos, relacionados con la importancia que tienen el BA para el buen desempeño reproductivo y productivo de l...

  13. [Animal experimentation, animal welfare and scientific research].

    Science.gov (United States)

    Tal, H

    2013-10-01

    Hundreds of thousands of laboratory animals are being used every year for scientific experiments held in Israel, mostly mice, rats, rabbits, guinea pigs, and a few sheep, cattle, pigs, cats, dogs, and even a few dozen monkeys. In addition to the animals sacrificed to promote scientific research, millions of animals slain every year for other purposes such as meat and fine leather fashion industries. While opening a front against all is an impossible and perhaps an unjustified task, the state of Israel enacted the Animal Welfare (Animal Experimentation) Law (1994). The law aims to regulate scientific animal experiments and to find the appropriate balance between the need to continue to perform animal experiments for the advancement of research and medicine, and at the same time to avoid unnecessary trials and minimize animal suffering. Among other issues the law deals with the phylogenetic scale according to which experimental animals should be selected, experiments for teaching and practicing, and experiments for the cosmetic industry. This article discusses bioethics considerations in animal experiments as well as the criticism on the scientific validity of such experiments. It further deals with the vitality of animal studies and the moral and legal obligation to prevent suffering from laboratory animals.

  14. IDENTIFIKASI DAGING BABI MENGGUNAKAN METODE PCR-RFLP GEN Cytochrome b DAN PCR PRIMER SPESIFIK GEN AMELOGENIN (Pork Identification Using PCR-RFLP of Cytochrome b Gene and Species Specific PCR of Amelogenin Gene

    Directory of Open Access Journals (Sweden)

    Yuny Erwanto

    2013-03-01

    Full Text Available A polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP and species specific PCR methods had been applied for identifying pork in mixture of meat. Pork sample in various levels (1, 3, 5 and 10% was prepared in mixture with beef, chicken and mutton. The primary CYTb1 and CYTb2 were designed in the mitochondrial cytochrome b b (cytochrome b gene and PCR successfully amplified fragments of 359 bp. To distinguish pig species existence, the amplified PCR products of mitochondrial DNA were cut by BseDI restriction enzyme. The result showed that pig mitochondrial DNA was cut into 131 and 228 bp fragments. A polymerase chain reaction (PCR method based on the nucleotide sequence variation in the amelogenin gene has been chosen for the specific identification of pork DNAs in mixture meat. The primers designed generated specific fragments of 353 and 312 bp length for pork. The specificity of the primary designed was tested on 4 animal species including pig, cattle, chicken and goat species. Analysis of experimental mixture meat demonstrated that 1% of raw pork tissues could be detected using PCR-RFLP with BseDI restriction enzyme but detection using species-specific PCR showed the cross reactivity to beef, chicken and mutton. The cytochrome b PCR-RFLP species identification assay yielded excellent results for identification of pig species. PCR-RFLP is a potentially reliable technique for detection of the existence of pork in animal food product for Halal authentication. Keywords: Pork identification, cytochrome b, amelogenin, polymerase chain reaction   ABSTRAK   Penelitian ini dilakukan untuk mengaplikasikan metode deteksi daging babi dalam campuan daging dengan sapi, kambing dan ayam melalui PCR-RFLP dan PCR dengan primer spesifik untuk babi. Level kontaminasi daging babi dibuat sebesar 1, 3, 5 dan 10% dari total daging dalam campuran. Metode PCR-RFLP menggunakan sepasang primer yaitu gen cytochrome b dari mitokondria yang

  15. Between and Animals

    Institute of Scientific and Technical Information of China (English)

    1994-01-01

    Animals are man’s best friends. Animals remind man of his own infancy. People and animals get on well with each other, so the world is bright and colorful. Animals are children’s close pals, too. Being on intimate terms with animals makes children more kind-hearted and sympathetic.

  16. Methylation-Specific PCR Unraveled

    Directory of Open Access Journals (Sweden)

    Sarah Derks

    2004-01-01

    Full Text Available Methylation‐specific PCR (MSP is a simple, quick and cost‐effective method to analyze the DNA methylation status of virtually any group of CpG sites within a CpG island. The technique comprises two parts: (1 sodium bisulfite conversion of unmethylated cytosine's to uracil under conditions whereby methylated cytosines remains unchanged and (2 detection of the bisulfite induced sequence differences by PCR using specific primer sets for both unmethylated and methylated DNA. This review discusses the critical parameters of MSP and presents an overview of the available MSP variants and the (clinical applications.

  17. A simple PCR-based procedure for plaque diagnosis Um método simples para o diagnóstico de peste por PCR

    Directory of Open Access Journals (Sweden)

    Nilma Cintra Leal

    1996-10-01

    Full Text Available Supernatant of boiled spleen saline-suspensions of Yersinia pestis experimentally infected animals were used as template for PCR amplification without DNA extraction. PCR sensitivity was enhanced by a second round of amplification (Nested. No amplification was observed from non-infected animals.Triturados de baços de animais infectados experimentalmente com Y. pestis, suspensos em salina foram fervidos e os sobrenadantes usados diretamente para amplificação do PCR sem prévia extração do DNA. O limiar de detecção poda ser aumentado por uma segunda etapa de amplificação (Nested-PCR. Não houve amplificação a partir das amostras dos animais não infectados usados como controle.

  18. High resolution positron tomography using PCR-I

    Energy Technology Data Exchange (ETDEWEB)

    Brownell, G.L.; Burnham, C.A.; Sandrew, B.; Elmaleh, D.R.; Livni, E.; Kizuka, H.

    1984-01-01

    PCR-I is a high resolution positron tomograph developed by the Physics Research Laboratory of the Massachusetts General Hospital to explore resolution limits of positron tomographs. PCR-I currently obtains images with 4.8 mm FWHM resolution at the center. Plane thickness may be varied between 5 and 10 mm. The instrument uses analog coding to obtain high resolution images without mechanical motion. This permits rapid dynamic imaging and gated cardiac imaging as well as conventional high resolution imaging. A series of studies has been carried out to demonstrate the ability of PCR-I to image structures in small animals. F-18 in the rat skeleton is clearly defined and various structures such as the spinal processes can be clearly resolved. A sequence of images at different spacing provides a three-dimensional reconstruction of the rat skeleton. Blood volume and palmitic acid have been imaged in the dog heart. Again, the sequence of images provides a clear delineation of the three dimensional nature of the blood pools and of the surrounding musculature. Blood flow, blood volume and glucose metabolism have been studied in the monkey brain. Structures within the brain of the Resus monkey can be clearly resolved. Increased activity resulting from induced seizures in the squirrel monkey have been observed and delineated. All of these studies indicate areas of future animal and clinical research using the high resolution tomograph, PCR-I.

  19. Animal Feeding Operations

    Science.gov (United States)

    ... What's this? Submit Button Healthy Water Home Animal Feeding Operations Recommend on Facebook Tweet Share Compartir On ... Concentrated Animal Feeding Operations (CAFOs) What are Animal Feeding Operations (AFOs)? According to the United States Environmental ...

  20. Animation of Antimicrobial Resistance

    Medline Plus

    Full Text Available ... The Food and Drug Administration's (FDA's) Center for Veterinary Medicine (CVM) produced a nine-minute animation explaining how ... and distributed as long as FDA's Center for Veterinary Medicine is cited as the corporate author. Animation Animation ...

  1. Animal welfare assessment

    Directory of Open Access Journals (Sweden)

    Vučinić Marijana

    2008-01-01

    Full Text Available The paper deals with animal welfare definitions and animal welfare assessment. Animal welfare is a prolonged mental state, resulting from how the animal experiences its environment over time. There are different methods for animal welfare assessment. The four basic criteria for animal welfare assessment are feeding, housing, health and appropriate behavior. Therefore, criteria used to assess animal welfare are not direct measures of the mental state but only parameters that need to be interpreted in terms of welfare. The immediate housing environment and feeding may influence animal welfare either positively, when most of the important requirements are respected, or negatively, when animals are exposed to various stress factors and unpleasant emotions that contribute to animal disease, injuries or inappropriate behavior. Therefore, animal welfare is a unique link between housing conditions, feeding and watering on one side, and animal health status and behavior on the other side.

  2. Animals and Medicine

    OpenAIRE

    Botting, Jack Howard; Botting, Regina; Morrison, Adrian R.

    2016-01-01

    Animals and Medicine: The Contribution of Animal Experiments to the Control of Disease offers a detailed, scholarly historical review of the critical role animal experiments have played in advancing medical knowledge. Laboratory animals have been essential to this progress, and the knowledge gained has saved countless lives - both human and animal. Unfortunately, those opposed to using animals in research have often employed doctored evidence to suggest that the practice has impeded medical p...

  3. Muestras y representatividad en vigilancia epidemiologica mediante sitios centinelas

    Directory of Open Access Journals (Sweden)

    Juan Samaja

    Full Text Available El artículo sostiene que las exigencias técnicas del muestreo para la vigilancia epidemiológica, exigen una revisión profunda de importantes conceptos de la Teoría de la Salud. En particular, es necesario hacer énfasis en las condiciones de vida, y, más específicamente, en los ambientes o contextos en que se desarrollan los procesos reproductivos de la vida social. Pero ambos campos temáticos exigen potenciar el acceso a datos más ricos que los que aportan las fuentes tradicionales. Este enfoque de la "vigilancia epidemiológica" exige una revisión de los tipos de muestras, y esto implica revisar las interpretaciones dominantes sobre los fundamentos lógicos de las inferencias a partir de muestras. Se torna necesario dejar atrás las muestras estadísticas (aún las estratificadas y promover procedimientos del tipo de los "sitios centinelas". Esta técnica, aplicada originariamente en sociedades con sistemas estadísticos deficitarios, puede desarrollarse para constituirse en un complemento substancial del monitoreo de condiciones de vida incluso en sociedades con buenos sistemas de información. El artículo propone transformar el concepto de "sitio centinela" incorporandole el requisito de la "representatividad cualitativa" mediante muestreos finalísticos sustentados en tipologías previas de las unidades espacio-poblacionales.

  4. Muestras y representatividad en vigilancia epidemiologica mediante sitios centinelas

    Directory of Open Access Journals (Sweden)

    Samaja Juan

    1996-01-01

    Full Text Available El artículo sostiene que las exigencias técnicas del muestreo para la vigilancia epidemiológica, exigen una revisión profunda de importantes conceptos de la Teoría de la Salud. En particular, es necesario hacer énfasis en las condiciones de vida, y, más específicamente, en los ambientes o contextos en que se desarrollan los procesos reproductivos de la vida social. Pero ambos campos temáticos exigen potenciar el acceso a datos más ricos que los que aportan las fuentes tradicionales. Este enfoque de la "vigilancia epidemiológica" exige una revisión de los tipos de muestras, y esto implica revisar las interpretaciones dominantes sobre los fundamentos lógicos de las inferencias a partir de muestras. Se torna necesario dejar atrás las muestras estadísticas (aún las estratificadas y promover procedimientos del tipo de los "sitios centinelas". Esta técnica, aplicada originariamente en sociedades con sistemas estadísticos deficitarios, puede desarrollarse para constituirse en un complemento substancial del monitoreo de condiciones de vida incluso en sociedades con buenos sistemas de información. El artículo propone transformar el concepto de "sitio centinela" incorporandole el requisito de la "representatividad cualitativa" mediante muestreos finalísticos sustentados en tipologías previas de las unidades espacio-poblacionales.

  5. Multi-primer target PCR for rapid identification of bovine DRB3 alleles.

    Science.gov (United States)

    Ledwidge, S A; Mallard, B A; Gibson, J P; Jansen, G B; Jiang, Z H

    2001-08-01

    Multi-primer target polymerase chain reaction (MPT-PCR) is a rapid method for the identification of specific BoLA-DRB3 alleles. In a single PCR reaction, the presence of two alleles associated with increased risk, DRB3.2*23 (DRB3*2701-2703, 2705-2707) and decreased risk, DRB3.2*16 (DRB3*1501, 1502), of mastitis in Canadian Holstein can be detected. Two outer primers amplify exon 2 of DRB3. Simultaneously, two inner, allele-specific primers amplify individual alleles. Initially, 40 cows previously typed by PCR-restriction fragment length polymorphism (PCR-RFLP) were genotyped using the multi-primer approach. An additional 30 cows were first genotyped by multi-primer target PCR, then by PCR-RFLP. All animals were correctly identified and there were no false positives. This technique can readily be modified to identify other BoLA alleles of interest.

  6. Development of a Real-time PCR test for porcine group A rotavirus diagnosis

    Directory of Open Access Journals (Sweden)

    Elizabeth C.M. Marconi

    2015-01-01

    Full Text Available Group A Rotavirus (RVA is one of the most common causes of diarrhea in humans and several animal species. A SYBR-Green Real-Time polymerase chain reaction (PCR was developed to diagnose RVA from porcine fecal samples, targeting amplification of a 137-bp fragment of nonstructural protein 5 (NSP5 gene using mRNA of bovine NADH-desidrogenase-5 as exogenous internal control. Sixty-five samples were tested (25 tested positive for conventional PCR and genetic sequencing. The overall agreement (kappa was 0.843, indicating 'very good' concordance between tests, presenting 100% of relative sensitivity (25+ Real Time PCR/25+ Conventional PCR and 87.5% of relative sensitivity (35- Real Time PCR/40- Conventional PCR. The results also demonstrated high intra- and inter-assay reproducibility (coefficient of variation ≤1.42%; thus, this method proved to be a fast and sensitive approach for the diagnosis of RVA in pigs.

  7. Animation of Antimicrobial Resistance

    Science.gov (United States)

    ... Animal & Veterinary Cosmetics Tobacco Products Animal & ... antimicrobial resistance both emerges and proliferates among bacteria. Over time, the use of antimicrobial drugs will result in the development ...

  8. Development of a real-time SYBR Green PCR assay for the rapid detection of Dermatophilus congolensis.

    Science.gov (United States)

    García, Alfredo; Martínez, Remigio; Benitez-Medina, José Manuel; Risco, David; Garcia, Waldo Luis; Rey, Joaquín; Alonso, Juan Manuel; Hermoso de Mendoza, Javier

    2013-01-01

    Methods such as real time (RT)-PCR have not been developed for the rapid detection and diagnosis of Dermatophilus (D.) congolensis infection. In the present study, a D. congolensis-specific SYBR Green RT-PCR assay was evaluated. The detection limit of the RT-PCR assay was 1 pg of DNA per PCR reaction. No cross-reaction with nucleic acids extracted from Pseudomonas aeruginosa, Mycobacterium tuberculosis, Staphylococcus aureus, or Austwickia chelonae was observed. Finally, the RT-PCR assay was used to evaluate clinical samples collected from naturally infected animals with D. congolensis. The results showed that this assay is a fast and reliable method for diagnosing dermatophilosis.

  9. Clostridium difficile ribotypes in humans and animals in Brazil

    Directory of Open Access Journals (Sweden)

    Rodrigo Otávio Silveira Silva

    2015-12-01

    Full Text Available Clostridium difficile is an emerging enteropathogen responsible for pseudomembranous colitis in humans and diarrhoea in several domestic and wild animal species. Despite its known importance, there are few studies aboutC. difficile polymerase chain reaction (PCR ribotypes in Brazil and the actual knowledge is restricted to studies on human isolates. The aim of the study was therefore to compare C. difficileribotypes isolated from humans and animals in Brazil. Seventy-six C. difficile strains isolated from humans (n = 25, dogs (n = 23, piglets (n = 12, foals (n = 7, calves (n = 7, one cat, and one manned wolf were distributed into 24 different PCR ribotypes. Among toxigenic strains, PCR ribotypes 014/020 and 106 were the most common, accounting for 14 (18.4% and eight (10.5% samples, respectively. Fourteen different PCR ribotypes were detected among human isolates, nine of them have also been identified in at least one animal species. PCR ribotype 027 was not detected, whereas 078 were found only in foals. This data suggests a high diversity of PCR ribotypes in humans and animals in Brazil and support the discussion of C. difficile as a zoonotic pathogen.

  10. Clostridium difficile ribotypes in humans and animals in Brazil.

    Science.gov (United States)

    Silva, Rodrigo Otávio Silveira; Rupnik, Maja; Diniz, Amanda Nádia; Vilela, Eduardo Garcia; Lobato, Francisco Carlos Faria

    2015-12-01

    Clostridium difficile is an emerging enteropathogen responsible for pseudomembranous colitis in humans and diarrhoea in several domestic and wild animal species. Despite its known importance, there are few studies about C. difficile polymerase chain reaction (PCR) ribotypes in Brazil and the actual knowledge is restricted to studies on human isolates. The aim of the study was therefore to compare C. difficile ribotypes isolated from humans and animals in Brazil. Seventy-six C. difficile strains isolated from humans (n = 25), dogs (n = 23), piglets (n = 12), foals (n = 7), calves (n = 7), one cat, and one manned wolf were distributed into 24 different PCR ribotypes. Among toxigenic strains, PCR ribotypes 014/020 and 106 were the most common, accounting for 14 (18.4%) and eight (10.5%) samples, respectively. Fourteen different PCR ribotypes were detected among human isolates, nine of them have also been identified in at least one animal species. PCR ribotype 027 was not detected, whereas 078 were found only in foals. This data suggests a high diversity of PCR ribotypes in humans and animals in Brazil and support the discussion of C. difficile as a zoonotic pathogen.

  11. Seeing the animal

    DEFF Research Database (Denmark)

    Harfeld, Jes; Cornou, Cécile; Kornum, Anna

    2016-01-01

    This article discusses the notion that the invisibility of the animalness of the animal constitutes a fundamental obstacle to change within current production systems. It is discussed whether housing animals in environments that resemble natural habitats could lead to a re-animalization...... of the animals, a higher appreciation of their moral significance, and thereby higher standards of animal welfare. The basic claim is that experiencing the animals in their evolutionary and environmental context would make it harder to objectify animals as mere bioreactors and production systems. It is argued...... that the historic objectification of animals within intensive animal production can only be reversed if animals are given the chance to express themselves as they are and not as we see them through the tunnel visions of economy and quantifiable welfare assessment parameters....

  12. Animal rights, animal minds, and human mindreading.

    Science.gov (United States)

    Mameli, M; Bortolotti, L

    2006-02-01

    Do non-human animals have rights? The answer to this question depends on whether animals have morally relevant mental properties. Mindreading is the human activity of ascribing mental states to other organisms. Current knowledge about the evolution and cognitive structure of mindreading indicates that human ascriptions of mental states to non-human animals are very inaccurate. The accuracy of human mindreading can be improved with the help of scientific studies of animal minds. However, the scientific studies do not by themselves solve the problem of how to map psychological similarities (and differences) between humans and animals onto a distinction between morally relevant and morally irrelevant mental properties. The current limitations of human mindreading-whether scientifically aided or not-have practical consequences for the rational justification of claims about which rights (if any) non-human animals should be accorded.

  13. Refining Animal Models to Enhance Animal Welfare

    Institute of Scientific and Technical Information of China (English)

    Patricia V.Turner

    2012-01-01

    The use of animals in research will be necessary for scientific advances in the basic and biomedical sciences for the foreseeable future.As we learn more about the ability of animals to experience pain,suffering,and distress,and particularly for mammals,it becomes the responsibility of scientists,institutions,animal caregivers,and veterinarians to seek ways to improve the lives of research animals and refine their care and use.Refinement is one of the three R's emphasized by Russell and Burch,and refers to modification of procedures to minimise the potential for pain,suffering and distress. It may also refer to procedures used to enhance animal comfort. This paper summarizes considerations for refinements in research animal.

  14. Detección sensible y específica de Mycobacterium tuberculosis a partir de muestras clínicas, mediante la amplificación de un elemento repetitivo de la familia REP13E12

    Directory of Open Access Journals (Sweden)

    Christian Baldeviano V

    2007-01-01

    Full Text Available Objetivos: Diseñar e implementar un método de PCR para la detección sensible y específica de M. tuberculosis, orientado al oportuno diagnóstico de la tuberculosis mediante su adecuada aplicación en muestras clínicas. Materiales y métodos: Se diseñaron oligonucleótidos específicos para la amplificación de un fragmento de 318pb luego de una alineación múltiple de secuencias REP13E12 de M. tuberculosis. Se realizó la estandarización del método de PCR y se evaluó la sensibilidad y especificidad diagnóstica utilizando muestras clínicas con diagnóstico baciloscópico y cultivo. Resultados: El REP13E12-PCR detectó hasta 100 fg de ADN genómico, fue específico para la detección del complejo M. tuberculosis y capaz de distinguirlos de micobacterias atípicas. Esta evaluación preliminar proporcionó 100% de sensibilidad y especificidad en muestras clínicas de pacientes con diagnóstico de TB. Conclusiones: La amplificación de REP13E12 mediante PCR es una alternativa para el diagnóstico rápido de pacientes con TB, especialmente en casos cuyo diagnóstico pueda ser dificultoso o poco claro mediante el uso de los métodos convencionales.

  15. Microbial Pollution Tracking of Dairy Farm with a Combined PCR-DGGE and qPCR Approach.

    Science.gov (United States)

    Xi, Xiaoxia; Zhang, Jiachao; Kwok, Laiyu; Huo, Dongxue; Feng, Shuzhen; Zhang, Heping; Sun, Tiansong

    2015-12-01

    Animal husbandry is a traditional industry with regional characteristic in the Inner Mongolia of China. Recent years, animal breeding has been one of the main pollution sources in this area, followed by domestic sewage and industrial wastewater. The pollution of livestock farm feces may accelerate the development of pathogens and antibiotic resistance genes which pose health risks to humans and animals. In present research, culture-independent molecular ecological methods based on DGGE combined with qPCR were used to investigate the pollution to surrounding environment with different degrees of livestock farm. The cluster analysis of DGGE patterns showed that the livestock farm feces from point pollution source flowed with wastewater discharge has resulted in an impacted range of at least 3000 m, but it did not cause pollution to residential water delivered from upstream of sewage drain outlet. qPCR results revealed that 5 common pathogens (Escherichia coli, Enterococcus, Staphylococcus aureus, Shigella, and Salmonella) presented decreased trend as the sampled distance from point pollution source increased. Also, qPCR assays of 10 common antibiotic resistance genes (tetO, tetL, rpp, rpoB, sul2, sulA, floR, yidY, mphA, and ermC) which cause resistance to tetracycline, rifampicin, fluoroquinolone, quinolone, and erythromycin have been found in the environmental samples. This study clearly indicates the livestock farm discharge pollutants contaminated to the surrounding environment. Our data have provided important information to pollution control in the future.

  16. Modelado de sistemas complejos mediante simulación basada en agentes y mediante dinámica de sistemas

    Directory of Open Access Journals (Sweden)

    LUIS R. IZQUIERDO

    2008-01-01

    Full Text Available Este trabajo compara dos técnicas de modelado de sistemas complejos: la simulación basada en agentes y la dinámica de sistemas. Esta comparación se lleva a cabo dentro del marco general del proceso de modelado científico. Los autores concluyen que la principal diferencia entre las dos metodologías se encuentra en el proceso de abstracción que cada una de ellas realiza para construir el modelo formal a partir del sistema complejo observado. Esta diferencia inicial se extiende a las restantes etapas del proceso de modelado científico. Finalmente, se indican los principales factores y las propiedades generales de un sistema complejo que hacen que una u otra técnica sea más relevante, aunque los autores destacan que, en la mayoría de los casos, modelizar un mismo sistema mediante las dos técnicas es la solución idónea.

  17. Detección de Cryptosporidium spp. en terneras de lecherías de la Región Metropolitana mediante Ziehl Neelsen y confirmada por inmunocromatografía y ensayo molecular Detection of Cryptosporidium spp. in calves by using a acid fast method and confirmed by immunochromatographic and molecular assays

    Directory of Open Access Journals (Sweden)

    P Muñoz

    2011-01-01

    Full Text Available Cryptosporidium causaría gran pérdida económica desde el punto de vista productivo, sobre todo en sistemas que involucren la crianza de bovinos afectando especialmente a animales menores de 30 días de edad con distintos grados de diarrea. El propósito de este estudio fue detectar Cryptosporidium spp. en muestras fecales de terneras diarreicas menores de un mes de edad en dos predios lecheros de la Región Metropolitana. Por primera vez en Chile se usó una prueba inmunocromatográfica (IC y una molecular (PCR para confirmar la observación microscópica de los ooquistes de Cryptosporidium spp. en la muestras fecales de bovinos estudiadas. En 49,8% (102/205 de las muestras fecales se observaron ooquistes de Cryptosporidium spp usando Ziehl Neelsen (ZN. De estas muestras positivas se seleccionaron al azar 58 para confirmar los diagnósticos mediante IC y todas resultaron también positivas. Diez muestras fecales ZN negativas también fueron confirmadas como negativas mediante IC. Mediante PCR en 37 de la 58 ZN positivas (64% se obtuvo un resultado positivo. La PCR también fue realizada en las diez muestras IC negativas sin obtener amplificación. La técnica molecular fue capaz de detectar muestras con menos ooquistes (10(4 ooquistes/ml en comparación con ZN (2 x 10(4 ooquistes/ml. Los resultados obtenidos permiten afirmar que la criptosporidiosis bovina sigue siendo una infección parasitaria de alta frecuencia en predios lecheros en la Región Metropolitana. Desde el punto de vista diagnóstico, la combinación de ZN con IC permitiría reducir la desventaja de ZN de ser una prueba operador dependiente. Se requiere de nuevos estudios que busquen incrementar el rendimiento de la PCR como prueba diagnóstica en la criptosporidiosis bovina. La implementación de pruebas moleculares también contribuye al estudio epidemiológico veterinario de esta parasitosis en una determinada área geográfica.From an animal production point of view

  18. Ian Ingram: Next Animals

    DEFF Research Database (Denmark)

    2015-01-01

    Ian Ingram: Next Animals is an exhibition catalogue presenting research on the work by Ian Ingram in relation to his exhibition Next Animals at Nikolaj Kunsthal in 2015.......Ian Ingram: Next Animals is an exhibition catalogue presenting research on the work by Ian Ingram in relation to his exhibition Next Animals at Nikolaj Kunsthal in 2015....

  19. Physics for Animation Artists

    Science.gov (United States)

    Chai, David; Garcia, Alejandro L.

    2011-01-01

    Animation has become enormously popular in feature films, television, and video games. Art departments and film schools at universities as well as animation programs at high schools have expanded in recent years to meet the growing demands for animation artists. Professional animators identify the technological facet as the most rapidly advancing…

  20. FARM ANIMAL WELFARE ECONOMICS

    Directory of Open Access Journals (Sweden)

    L.T. CZISZTER

    2013-07-01

    Full Text Available This paper reviews the literature regarding the economics of the farm animal welfare. The following issues are addressed: productions costs and savings of the animal welfare regulations, benefits of improved animal welfare, and consumers’ willingness to pay for animal-friendly products.

  1. Climático: evaluación mediante modelos

    Directory of Open Access Journals (Sweden)

    P. Ruiz-Benito

    2013-01-01

    Full Text Available Los bosques son ecosistemas fundamentales en la generación de servicios ecosistémicos y, por tanto, para el bienestar humano. El cambio global (incluyendo cambio climático y cambios en el uso del suelo puede, sin embargo, alterar la dinámica y el funcionamiento de los ecosistemas, afectando al futuro suministro de servicios ecosistémicos. La vulnerabilidad frente al cambio global depende de la exposición (magnitud del cambio, la sensibilidad (susceptibilidad al cambio, y la capacidad de adaptación (habilidad para ajustarse al cambio de las especies. En el presente trabajo presentamos diversas aproximaciones de modelización que permiten analizar los diferentes componentes de la vulnerabilidad, e incluimos ejemplos desarrollados para bosques de la península Ibérica. A pesar de estos avances, la evidencia empírica y teórica para integrar los impactos potenciales (i.e. incluyendo la exposición y la sensibilidad y la capacidad de adaptación de las especies, es escasa. Por ello, para una adecuada evaluación sería necesario me-jorar el conocimiento existente sobre la sensibilidad y capacidad de adaptación de las especies y su respuesta frente a cambios ambientales extremos (por ejemplo, mediante redes de seguimiento a largo plazo, integrando adecuadamente la información obtenida en modelos que incluyan procesos basados en diferentes niveles de organización biológica, desde procesos fisiológicos a modelos agregados de distribución de especies.

  2. Carotenoids in Marine Animals

    Directory of Open Access Journals (Sweden)

    Takashi Maoka

    2011-02-01

    Full Text Available Marine animals contain various carotenoids that show structural diversity. These marine animals accumulate carotenoids from foods such as algae and other animals and modify them through metabolic reactions. Many of the carotenoids present in marine animals are metabolites of β-carotene, fucoxanthin, peridinin, diatoxanthin, alloxanthin, and astaxanthin, etc. Carotenoids found in these animals provide the food chain as well as metabolic pathways. In the present review, I will describe marine animal carotenoids from natural product chemistry, metabolism, food chain, and chemosystematic viewpoints, and also describe new structural carotenoids isolated from marine animals over the last decade.

  3. Ethics in Animal Experimentation

    Directory of Open Access Journals (Sweden)

    Yusuf Ergun

    2010-08-01

    Full Text Available Experimental animals are frequently used to obtain information for primarily scientific reasons. In the present review, ethics in animal experimentation is examined. At first, the history of animal experimentation and animal rights is outlined. Thereafter, the terms in relation with the topic are defined. Finally, prominent aspects of 3Rs constituting scientific and ethical basis in animal experimentation are underlined. [Archives Medical Review Journal 2010; 19(4.000: 220-235

  4. Ethics in Animal Experimentation

    OpenAIRE

    Yusuf Ergun

    2010-01-01

    Experimental animals are frequently used to obtain information for primarily scientific reasons. In the present review, ethics in animal experimentation is examined. At first, the history of animal experimentation and animal rights is outlined. Thereafter, the terms in relation with the topic are defined. Finally, prominent aspects of 3Rs constituting scientific and ethical basis in animal experimentation are underlined. [Archives Medical Review Journal 2010; 19(4.000): 220-235

  5. Animal Images and Metaphors in Animal Farm

    Directory of Open Access Journals (Sweden)

    Ping Sun

    2015-05-01

    Full Text Available In literary works animal images are frequently used as the “source domain” of a metaphor to disclose the natures of the “target domain”, human beings. This is called “cross-domain mapping” or “conceptual metaphor” in cognitive linguistics, which is based on the similar qualities between animals and human beings. Thus the apparent descriptions of the animals are really the deep revelations of the human beings. Animal Farm is one exemplary product of this special expressing way. Diversified animal images are intelligently used by George Orwell to represent the people, so all the characters are animals in appearance, but humans in nature. Starting from the animal images and then the conceptual metaphors, readers can perceive a fresh understanding of this classical book. In this novel, three conceptual metaphors are identified and the special findings can be illustrated as the following: Firstly, the whole story of the animals represents the history and politics of the Soviet Union. Secondly, the pigs symbolize the authorities of the society. Thirdly, the names of the characters in the novel reveal their identities.

  6. Chirurgia della valvulopatia mitralica mediante Port Access. Esperienza di due centri.

    OpenAIRE

    Iorio, Francesco

    2014-01-01

    La chirurgia mininvasiva della mitrale mediante Port Access, risulta una delle maggiori innovazioni in cardiochirurgia in grado di migliorare gli outcome a breve medio e lungo termine se paragonata alla chirurgia convenzionale. In questo studio sono sono prese in esame le esperienze di due centri di eccellenza che hanno condotto tali interventi mediante approcci tecnici intraoperatori diversi per il clampaggio aortico: l'Endoclamp ed il clampaggio aortico transtoracico. Vengono altresì discus...

  7. Detection of Mycobacterium bovis in bovine and bubaline tissues using nested-PCR for TbD1.

    Directory of Open Access Journals (Sweden)

    Cristina P Araújo

    Full Text Available In the present study, a nested-PCR system, targeting the TbD1 region, involving the performance of conventional PCR followed by real-time PCR, was developed to detect Mycobacterium bovis in bovine/bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. In terms of analytical sensitivity, the DNA of M. bovis AN5 was detected up to 1.56 ng with conventional PCR, 97.6 pg with real-time PCR, and 1.53 pg with nested-PCR in the reaction mixture. The nested-PCR exhibited 100% analytical specificity for M. bovis when tested with the DNA of reference strains of environmental mycobacteria and closely-related Actinomycetales. A clinical sensitivity value of 76.0% was detected with tissue samples from animals that exhibited positive results in the comparative intradermal tuberculin test (CITT, as well as from those with lesions compatible with tuberculosis (LCT that rendered positive cultures. A clinical specificity value of 100% was detected with tissue samples from animals with CITT- results, with no visible lesions (NVL and negative cultures. No significant differences were found between the nested-PCR and culture in terms of detecting CITT+ animals with LCT or with NVL. No significant differences were recorded in the detection of CITT- animals with NVL. However, nested-PCR detected a significantly higher number of positive animals than the culture in the group of animals exhibiting LCT with no previous records of CITT. The use of the nested-PCR assay to detect M. bovis in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis.

  8. Molecular diagnosis of canine visceral leishmaniasis: identification of Leishmania species by PCR-RFLP and quantification of parasite DNA by real-time PCR.

    Science.gov (United States)

    Quaresma, Patrícia Flávia; Murta, Silvane Maria Fonseca; Ferreira, Eduardo de Castro; da Rocha-Lima, Ana Cristina Vianna Mariano; Xavier, Ana Amélia Prates; Gontijo, Célia Maria Ferreira

    2009-09-01

    The efficacies of polymerase chain reaction (PCR) procedures for the diagnosis of canine visceral leishmaniasis (CVL), and of PCR-restriction fragment length polymorphism (RFLP) analysis for the identification of Leishmania species, have been assessed. Quantitative real-time PCR employing a SYBR Green dye-based system was standardised for the quantification of Leishmania kDNA minicircles. Skin, peripheral blood and bone marrow samples collected from 217 dogs, asymptomatic or symptomatic for CVL, were analysed. The PCR method, which was based on the amplification of a 120 bp kDNA fragment conserved across Leishmania species, was able to detect the presence in clinical samples of protozoan parasite DNA in amounts as low as 0.1 fg. Bone marrow and skin samples proved to be more suitable than peripheral blood for the detection of Leishmania by PCR and presented positive indices of 84.9% and 80.2%, respectively. PCR-RFLP analysis indicated that 192 of the PCR-positive dogs were infected with Leishmania infantum chagasi, whilst L. braziliensis was identified in two other animals. Quantitative PCR revealed that bone marrow samples from dogs presenting positive conventional tests contained a higher number of copies of Leishmania kDNA than peripheral blood, although no significant differences were detected between symptomatic and asymptomatic dogs in terms of parasite load. This study demonstrates that PCR can be used for the detection of Leishmania in clinical samples derived from naturally infected dogs, and that PCR-RFLP represents a rapid and sensitive tool for the identification of Leishmania species. Additionally, qPCR is effective in quantifying Leishmania DNA load in clinical samples.

  9. Detection of Mycobacterium avium subsp. paratuberculosis in Milk from Clinically Affected Cows by PCR and culture

    DEFF Research Database (Denmark)

    Giese, Steen Bjørck; Ahrens, Peter

    1999-01-01

    -negative on intestinal mucosa, but culture-positive in milk, and both faeces and milk were negative in culture and PCR from 2 cows. In conclusion the presence of M. a. paratuberculosis could be detected in raw milk by PCR but cultivation of milk was more sensitive in detecting the organism.......Milk and faecal samples from cows with clinical symptoms of paratuberculosis were examined for the presence of Mycobacterium avium subsp.paratuberculosis (M. a. paratuberculosis) by culture and PCR. M. a. paratuberculosis was isolated in varied numbers from faeces or intestinal mucosa in 8 of 11...... animals. In milk from 5 cows (all faecal culture-positive) we cultivated a few colonies of M. a. paratuberculosis (less than 100 CFU per mi). Milk samples from 2 cows were PCR-positive (both animals were faecal culture-positive, and 1 cow was milk culture positive). One cow was culture...

  10. Desarrollo de la lectura mediante estratégias integradoras

    Directory of Open Access Journals (Sweden)

    Solé, Maira

    2005-06-01

    Full Text Available La lectura y la escritura son procesos que cada día ameritan nuevos cambios y transformaciones. La propuesta de un Proyecto Pedagógico Integrador, (Fraca 2003 desarrollado con éxito en algunas instituciones venezolanas, se perfila como una alternativa significativa para el desarrollo de estos elementos. La idea o núcleo central es la integración de las diferentes asignaturas curriculares y lograr una globalización partiendo de sus objetivos y contenidos programáticos. El eje pedagógico integrador le permite al docente, evidenciar con mayor prontitud los resultados mediante actividades prácticas de lectura y escritura. Así mismo combina elementos claves del aprendizaje ausbeliano: información previa, información nueva y construcción de la información definitiva o integrada. La puesta en ejecución de las estrategias integradoras, en esta ocasión por maestros en formación (UNEG, a diferentes niños de escuelas del Estado Bolívar (Venezuela, certificando cómo la lectura y la escritura pueden tener un espacio ideal y significativo en la instrucción actual. Solo se necesita la intención, creatividad, dinamismo e ingenio. The reading and the writing plows processes that every day they require new changes and transformations. The proposal of an Integrative Pedagogic Project, (Fraca 2003 developed with success in some Venezuelan institutions; it is profiled like a significant alternative for the development of these elements. The idea or central nucleus is the integration of the different curricular subjects and to achieve a globalization leaving of its objectives and programmatic contents. The integrative pedagogic axis allows to the educational one, to evidence with more readiness the results by means of practical activities of reading and it notarizes. Likewise it combines key elements of the learning ausbeliano: previous information, new information and construction of the definitive or integrated information. The operation of

  11. de teorías mediante un estudio de caso

    Directory of Open Access Journals (Sweden)

    Verónica Alonso Jiménez

    2008-01-01

    Full Text Available La democracia representativa como forma de gobierno, implica que el poder se ejerce por personas, que elegidas por el pueblo, actúan en su nombre y representación. El modo de participación en la elección y la manera como éstas se convierten en cargos públicos, requiere del dise- ño de instituciones que sistematicen dicha participación. La república de tipo presidencial, es una modalidad del gobierno electivo y popular, cuyo titular es el jefe del ejecutivo, electo por el pueblo o sus representantes, en donde el ejercicio del poder es limitado y mantiene un régimen de responsabilidades políticas. El diseño institucional que le corresponde a esta forma de gobierno es la parcelación del poder pú- blico en tres: poder ejecutivo, poder legislativo y poder judicial. Las ventajas políticas de este esquema es que la división de poderes, neutraliza el riesgo de caer en el autoritarismo, al impedir que el poder se concentre. La división de poderes es un dispositivo de restricción de facultades de los órganos estatales, por lo que no existe superioridad jerárquica entre los poderes, al contrario, cada órgano tiene bien delimitadas sus funciones y atribuciones, las que están reguladas por un marco jurídico común llamado Constitución. Y la división de poderes contribuye a mantener el equilibrio entre estos, mediante el llamado sistema de “pesos y contrapesos”. El presente trabajo se centra en el estudio de la Cámara de Diputados, considerada como una de las parcelas en las que está divida la autoridad del Estado, cuyo objetivo es la validación de los modelos teóricos en la tipificación del Congreso y la comprensión de las variables internas y externas que influyen en el comportamiento legislativo.

  12. Identificación por reacción en cadena de la polimerasa (PCR) de microorganismos presentes en las infecciones orofaciales odontogénicas

    OpenAIRE

    Ghersi Miranda, Hugo Dante Francisco; Facultad de Estomatología, Universidad Peruana Cayetano Heredia. Lima,; Inga Peña, Rocío de María; Facultad de Ciencias y Filosofía. Universidad Peruana Cayetano Heredia. Lima,

    2014-01-01

    El objetivo del presente trabajo fue identificar la presencia de las bacterias, Porphyromonas gingivalis, Tannerella forsythensis y Treponema denticola, denominado complejo rojo; en grupo de tres, en parejas o sola una especie, presentes en las infecciones orofaciales odontogénicas (IOFO), mediante el método de la Reacción de la Polimerasa Reversa (PCR), en las muestras de abscesos de los pacientes que acudieron al Servicio de Odontoestomatologia del Hospital Nacional Cayetano Heredia y a la ...

  13. Two-temperature PCR for Microfluidics

    KAUST Repository

    Kodzius, Rimantas

    2010-05-01

    Since its invention in 1983, polymerase chain reaction (PCR) has been the method of choice for DNA amplification. Successful PCR depends on the optimization of several parameters, which is a cumbersome task due to the many variables (conditions and compon

  14. RETHINKING THE ANIMATE, RE-ANIMATING THOUGHT

    Directory of Open Access Journals (Sweden)

    Tim Ingold

    2013-12-01

    Full Text Available Animism is often described as the imputation of life to inert objects. Such imputation is more typical of people in western societies who dream of finding life on other planets than of indigenous peoples to whom the label of animism has classically been applied. These peoples are united not in their beliefs but in a way of being that is alive and open to a world in continuous birth. In this animic ontology, beings do not propel themselves across a ready-made world but rather issue forth through a world-in-formation, along the lines of their relationships. To its inhabitants this weather-world, embracing both sky and earth, is a source of astonishment but not surprise. Re-animating the ‘western’ tradition of thought means recovering the sense of astonishment banished from offi cial science.

  15. Effectivity of PCR and AGID methods to detect of enzootic bovine leukosis in Indonesia

    Directory of Open Access Journals (Sweden)

    Saepulloh M

    2015-03-01

    Full Text Available Enzootic Bovine Leucosis (EBL is one of viral diseases in cattle caused by bovine leukemia virus (BLV, from Retroviridae. The virus can be detected using severals methods such as Polymerase Chain Reaction (PCR, while antibody can be detected using Agar Gel Immunodifussion (AGID. The aim of this experiment was to study the effectivity of PCR and AGID methods to detect enzootic bovine leukosis virus in Indonesia. Samples of peripheral blood leukocyte (PBL were collected from cattles those with and without showing clinical signs. A total of 307 blood and serum samples were tested against BLV using PCR and AGID tests, while 21 semen samples which were from similar animals for blood collection were collected only for PCR test. The results indicated that twelve cattles have positive results with PCR test in PBL, but from those cattles only seven were positive with AGID. On the other hand, the PCR did not detect EBL in 21 bovine semen samples tested, although one sample gave positive result with PCR in PBL. This results indicated that PCR method from blood samples was more sensitive than that AGID method. The PCR detection was also more sensitive for PBL than that for semen samples

  16. Use of pan-hemoplasma PCR for screening horses highly exposed to tick bites from southern Brazil

    Directory of Open Access Journals (Sweden)

    Thállitha Samih Wischral Jayme Vieira

    2015-02-01

    Full Text Available Mycoplasma sp. are hemotropic bacteria of red blood cells and the causative agent of hemoplasmosis. Diagnosis of infection is reached by polymerase chain reaction (PCR and real-time PCR (qPCR of the 16S RNA gene of this bacteria. The aim of this study was to screen horses for Mycoplasma sp. Infection using a pan-hemoplasma SYBR green qPCR assay. A total of 198 blood samples from horses were collected, DNA extracted and pan-hemoplasma qPCR performed. All samples were negative by qPCR. To verify the presence of amplifiable DNA, PCR for glyceraldehyde-3-phosphate dehydrogenase gene was performed on all samples. This study was part of an active surveillance program, which is critical for monitoring animal health status, particularly in horses.

  17. "Name" that Animal

    Science.gov (United States)

    Laird, Shirley

    2010-01-01

    In this article, the author describes a texture and pattern project. Students started by doing an outline contour drawing of an animal. With the outline drawn, the students then write one of their names to fit "inside" the animal.

  18. Animation of Antimicrobial Resistance

    Science.gov (United States)

    ... Veterinary Safety & Health Antimicrobial Resistance Animation of Antimicrobial Resistance Share Tweet Linkedin Pin it More sharing options ... produced a nine-minute animation explaining how antimicrobial resistance both emerges and proliferates among bacteria. Over time, ...

  19. Animation of Antimicrobial Resistance

    Medline Plus

    Full Text Available ... Animal & Veterinary Cosmetics Tobacco Products Animal & ... antimicrobial resistance both emerges and proliferates among bacteria. Over time, the use of antimicrobial drugs will result in the development ...

  20. DIAGNÓSTICO ANTENATAL DE SÍFILIS CONGÉNITA POR PCR EN LÍQUIDO AMNIÓTICO Y SUS POSIBLES APLICACIONES

    OpenAIRE

    Yamamoto C,Masami; Lizama M.,Luis; Valenzuela H.,Patricia; Carrillo T.,Jorge; Insunza F,Alvaro; Prado D.,Priscila

    2005-01-01

    Se presenta un caso clínico de sífilis congénita diagnosticada antenatalmente mediante el uso de la reacción de la polimerasa en cadena (PCR) en líquido amniótico. La PCR permitiría identificar la espiroqueta en diferentes medios, como en sangre, líquido amniótico y líquido céfalo-raquídeo. Deberán desarrollarse nuevos protocolos para poder probar la efectividad de los tratamientos en base a esta técnica diagnóstica

  1. DETECTING LOW DENSITY LIPOPROTEIN RECEPTOR MUTANT GENE OF RABBIT BY PCR

    Institute of Scientific and Technical Information of China (English)

    Liu Enqi; Zhao Sihai; Chen Zhenglan; Yang Penghui

    2006-01-01

    Objective Watanabe Heritable Hyperlipidaemic (WHHL) rabbits with low density lipoprotein receptor (LDLr) gene mutation have provided unprecedented opportunities for the study of human atherosclerosis, in order to confirm LDL receptor gene status in rabbits, we developed a simple PCR technique to detect LDL mutations in rabbits. Methods Rabbits genomic DNA were extracted from ear biopsy, and amplified by PCR to detect 12 bp deletion mutation in WHHL rabbits. PCR products were directly digested with BglⅠ, and then applied to polyacrylamide gel electrophoresis. Results PCR products from homozygous LDLr +/+ rabbits generated 2 bands of 212 and 94 bp after BglⅠ digestion, LDLr +/- rabbits generated 3 bands (294, 212, and 94 bp), LDLr -/- animals, however, generated only 1 product (294 bp). Conclusion This modified PCR method is simple and reliable.

  2. Animal violence demystified

    NARCIS (Netherlands)

    Natarajan, Deepa; Caramaschi, Doretta

    2010-01-01

    Violence has been observed in humans and animals alike, indicating its evolutionary/biological significance. However, violence in animals has often been confounded with functional forms of aggressive behavior. Currently, violence in animals is identified primarily as either a quantitative behavior (

  3. I like animals

    Institute of Scientific and Technical Information of China (English)

    官健

    2008-01-01

    @@ Animals are our friends.We should protect them and we mustn't hurtthem. Do you like animals?My answer is"yes".Maybe you may ask me why.I will tell you they are very lovely.I like many animals,such as pandas,monkeys and elephants.

  4. Animation of Antimicrobial Resistance

    Medline Plus

    Full Text Available ... Skip to common links HHS U.S. Department of Health and Human Services U.S. Food and Drug Administration ... Tobacco Products Animal & Veterinary Home Animal & Veterinary Safety & Health Antimicrobial Resistance Animation of Antimicrobial Resistance Share Tweet ...

  5. Diagnóstico rápido y efectivo de brucelosis bovina en sangre, mediante una reacción en cadena de la polimerasa doble

    OpenAIRE

    Víctor Eustorgio Aguirre Arzola; Mónica Alvarado González; José Luís Ibave González; Marisela Leal Hernández; Efrén Díaz Aparicio; Guadalupe Virginia Nevárez Moorillón; Francisco Javier Solís Martínez; Sigifredo Arévalo Gallegos; Blanca Estela Rivera Chavira

    2008-01-01

    La brucelosis es una de las enfermedades infecciosas más importantes del ganado y representa una barrera para la importación y exportación de productos lácteos y cárnicos. En México, el diagnóstico se realiza mediante las pruebas serológicas de tarjeta, rivanol y fijación del complemento. Los métodos moleculares, como la reacción en cadena de la polimerasa (PCR), son herramientas rápidas y específicas para el diagnóstico de la enfermedad. En el presente trabajo se desarrolló el diagnóstico de...

  6. Determination of processed animal proteins, including meat and bone meal, in animal feed.

    Science.gov (United States)

    Gizzi, Giséile; von Holst, Christoph; Baeten, Vincent; Berben, Gilbert; van Raamsdonk, Leo

    2004-01-01

    An intercomparison study was conducted to determine the presence of processed animal proteins (PAPs), including meat and bone meal (MBM) from various species, in animal feed. The performances of different methods, such as microscopy, polymerase chain reaction (PCR), immunoassays, and a protocol based on iquid chromatography (LC), were compared. Laboratories were asked to analyze for PAPs from all terrestrial animals and fish (total PAPs); mammalian PAPs; ruminant PAPs; and porcine PAPs. They were free to use their method of choice. In addition, laboratories using microscopy were asked to determine the presence of PAPs from terrestrial animals, which is applicable only to microscopy. For total PAPs microscopy, LC and some immunoassays showed sufficient results at a concentration as low as 0.1% MBM in the feed. In contrast, PCR was not fit for purpose. In differentiating between MBM from terrestrial animals and fishmeal, microscopy detected 0.5% of terrestrial MBM in feed in the presence of 5% fishmeal, but was less successful when the concentration of MBM from terrestrial animals was 0.1%. The animal-specific determination of MBM from mammals or, more specifically from either ruminants or pigs, by PCR showed poor results, as indicated by a high number of false-positive and false-negative results. The only PCR method that scored quite well was applied by a member of the organizer team of the study. Immunoassays scored much better than PCR, showing sufficient sensitivity but some deficiency in terms of specificity. The results also demonstrated that the reliable determination of MBM from ruminants has not been resolved, especially for low concentrations of MBM (0.1%) in feed. Comparison of the results for mammalian MBM from all methods indicated that, for control purposes, the immunoassay method, especially when applied as dipsticks, could be used as a rapid screening method combined with microscopy to confirm the positive samples. However, implementation of such a

  7. Evaluation of combined high-efficiency DNA extraction and real-time PCR for detection of Mycobacterium avium subsp. paratuberculosis in subclinically infected dairy cattle: comparison with faecal culture, milk real-time PCR and milk ELISA

    Directory of Open Access Journals (Sweden)

    Logar Katarina

    2012-05-01

    -shedders in herds. Identification of these shedding animals is extremely important for prevention of the spread of Map infection in an animal population. Due to the relatively high sensitivity and specificity of HYDEqPCR, it can be applied to test for Map at the herd or individual level, regardless of animal age or production stage. HYDEqPCR will allow early detection and control of Map in any population at risk.

  8. Real-time PCR in Food Science: PCR Diagnostics.

    Science.gov (United States)

    Rodriguez-Lazaro, David; Cook, Nigel; Hernandez, Marta

    2013-01-01

    A principal consumer demand is a guarantee of the safety and quality of food. The presence of foodborne pathogens and their potential hazard, the use of genetically modified organisms (GMOs) in food production, and the correct labelling in foods suitable for vegetarians are among the subjects where society demands total transparency. The application of controls within the quality assessment programmes of the food industry is a way to satisfy these demands, and is necessary to ensure efficient analytical methodologies are possessed and correctly applied by the Food Sector. The use of real-time PCR has become a promising alternative approach in food diagnostics. It possesses a number of advantages over conventional culturing approaches, including rapidity, excellent analytical sensitivity and selectivity, and potential for quantification. However, the use of expensive equipment and reagents, the need for qualified personnel, and the lack of standardized protocols are impairing its practical implementation for food monitoring and control.

  9. Animal models of dementia

    DEFF Research Database (Denmark)

    Olsson, I. Anna S.; Sandøe, Peter

    2011-01-01

    This chapter aims to encourage scientists and others interested in the use of animal models of disease – specifically, in the study of dementia – to engage in ethical reflection. It opens with a general discussion of the moral acceptability of animal use in research. Three ethical approaches...... are here distinguished. These serve as points of orientation in the following discussion of four more specific ethical questions: Does animal species matter? How effective is disease modelling in delivering the benefits claimed for it? What can be done to minimize potential harm to animals in research? Who...... bears responsibility for the use of animals in disease models?...

  10. DETECCION DE Brucella abortus POR PCR EN MUESTRAS DE SANGRE Y LECHE DE VACUNOS

    Directory of Open Access Journals (Sweden)

    Xiomara Mosquera C

    2008-12-01

    Full Text Available Objetivo. Evaluar el uso de la Reacción en Cadena de la Polimerasa (PCR para la detección de Brucella abortus en muestras de sangre y leche de vacunos. Materiales y métodos. Este estudio de tipo descriptivo fue realizado durante los años 2004 y 2005. Se analizaron 136 animales de tres fincas localizadas en el municipio de Durania, Norte de Santander, Colombia. Se evaluó la presencia de anticuerpos en la leche mediante la prueba del anillo (PAL. Se amplificó el fragmento de 223pb del gen BCSP31. Se emplearon los cebadores B4 y B5 de la región interna de la secuencia del gen BCSP31 (GenBank, número M20404. Resultados. En aquellos animales positivos se obtuvo una muestra de sangre y leche para el análisis por PCR, la sangre no fue analizada por serología. Se evaluaron diferentes métodos de extracción de ADN. Se encontró que un 13.2% (18/136 de las muestras de leche fueron positivas a la PAL. Se analizaron 33 muestras de leche negativas por PAL de las cuales el 30.3% (10/33 resultaron positivas por PCR. Al analizar las muestras de sangre de los animales positivos por PAL el 94.1% (16/17 fueron positivas por PCR, mientras que el 47% (8/17 de las muestras de leche positivas por PAL, fueron positivas por PCR. Conclusiones. Se demostró la amplificación de un fragmento de ADN de Brucella abortus en muestras de sangre y leche de vacunos. Los resultados preliminares demostraron que es posible usar PCR como prueba diagnóstica de brucelosis en Colombia.

  11. Real-Time PCR (qPCR) Primer Design Using Free Online Software

    Science.gov (United States)

    Thornton, Brenda; Basu, Chhandak

    2011-01-01

    Real-time PCR (quantitative PCR or qPCR) has become the preferred method for validating results obtained from assays which measure gene expression profiles. The process uses reverse transcription polymerase chain reaction (RT-PCR), coupled with fluorescent chemistry, to measure variations in transcriptome levels between samples. The four most…

  12. [Animal experimentation in Israel].

    Science.gov (United States)

    Epstein, Yoram; Leshem, Micah

    2002-04-01

    In 1994 the Israeli parliament (Knesset) amended the Cruelty to Animals Act to regulate the use of experimental animals. Accordingly, animal experiments can only be carried out for the purposes of promoting health and medical science, reducing suffering, advancing scientific research, testing or production of materials and products (excluding cosmetics and cleaning products) and education. Animal experiments are only permitted if alternative methods are not possible. The National Board for Animal Experimentation was established to implement the law. Its members are drawn from government ministries, representatives of doctors, veterinarians, and industry organizations, animal rights groups, and academia. In order to carry out an animal experiment, the institution, researchers involved, and the specific experiment, all require approval by the Board. To date the Board has approved some 35 institutions, about half are public institutions (universities, hospitals and colleges) and the rest industrial firms in biotechnology and pharmaceutics. In 2000, 250,000 animals were used in research, 85% were rodents, 11% fowls, 1,000 other farm animals, 350 dogs and cats, and 39 monkeys. Academic institutions used 74% of the animals and industry the remainder. We also present summarized data on the use of animals in research in other countries.

  13. Anchored PCR (A-PCR):A new method for chromosome walking

    Institute of Scientific and Technical Information of China (English)

    CHEN Bojun; SUN Chao; WANG Yong; HU Yuanlei; LIN Zhongping

    2004-01-01

    @@ PCR-based techniques are most popular methods for isolation of DNA sequences flanking a known region.Such techniques published to date mainly include three types: inverse PCR (IPCR)[1-3], ligation-mediated PCR (LM-PCR)[4-9] and randomly primed PCR (RP-PCR)[10-12].IPCR was the first method developed for this kind of purpose. However, it is now rarely used because of the difficulty in finding suitable restriction sites in the target region or poor circularization of the template molecule.LM-PCR and RP-PCR are more frequently used nowadays, yet they also have some limitations. For example,LM-PCR depends on restriction sites within a reasonable distance in the flanking regions, while the amplified products of RP-PCR are generally small (<1 kb). Moreover, both methods often result in excessive amplification of non-specific molecules, which greatly reduces their efficiencies in obtaining sequences of interest. To resolve these problems, some new strategies have emerged in the past few years, such as Vectorette-PCR[6], biotin-capture PCR[7], TAIL-PCR[l2] and T-linker PCR[9]. These improved methods are more efficient than their old versions;however, most of them are still limited by restriction digestion or ligation. Although the intervening steps are avoided in TAIL-PCR, the amplified fragments are often small because of the use of random primers.

  14. PCR Conditions for 16S Primers for Analysis of Microbes in the Colon of Rats.

    Science.gov (United States)

    Guillen, I A; Camacho, H; Tuero, A D; Bacardí, D; Palenzuela, D O; Aguilera, A; Silva, J A; Estrada, R; Gell, O; Suárez, J; Ancizar, J; Brown, E; Colarte, A B; Castro, J; Novoa, L I

    2016-09-01

    The study of the composition of the intestinal flora is important to the health of the host, playing a key role in maintaining intestinal homeostasis and the evolution of the immune system. For these studies, various universal primers of the 16S rDNA gene are used in microbial taxonomy. Here, we report an evaluation of 5 universal primers to explore the presence of microbial DNA in colon biopsies preserved in RNAlater solution. The DNA extracted was used for the amplification of PCR products containing the variable (V) regions of the microbial 16S rDNA gene. The PCR products were studied by restriction fragment length polymorphism (RFLP) analysis and DNA sequence, whose percent of homology with microbial sequences reported in GenBank was verified using bioinformatics tools. The presence of microbes in the colon of rats was quantified by the quantitative PCR (qPCR) technique. We obtained microbial DNA from rat, useful for PCR analysis with the universal primers for the bacteria 16S rDNA. The sequences of PCR products obtained from a colon biopsy of the animal showed homology with the classes bacilli (Lactobacillus spp) and proteobacteria, normally represented in the colon of rats. The proposed methodology allowed the attainment of DNA of bacteria with the quality and integrity for use in qPCR, sequencing, and PCR-RFLP analysis. The selected universal primers provided knowledge of the abundance of microorganisms and the formation of a preliminary test of bacterial diversity in rat colon biopsies.

  15. Universal reverse-transcriptase real-time PCR for infectious hematopoietic necrosis virus (IHNV)

    Science.gov (United States)

    Purcell, Maureen K.; Thompson, Rachel L.; Garver, Kyle A.; Hawley, Laura M.; Batts, William N.; Sprague, Laura; Sampson, Corie; Winton, James R.

    2013-01-01

    Infectious hematopoietic necrosis virus (IHNV) is an acute pathogen of salmonid fishes in North America, Europe and Asia and is reportable to the World Organization for Animal Health (OIE). Phylogenetic analysis has identified 5 major virus genogroups of IHNV worldwide, designated U, M, L, E and J; multiple subtypes also exist within those genogroups. Here, we report the development and validation of a universal IHNV reverse-transcriptase real-time PCR (RT-rPCR) assay targeting the IHNV nucleocapsid (N) gene. Properties of diagnostic sensitivity (DSe) and specificity (DSp) were defined using laboratory-challenged steelhead trout Oncorhynchus mykiss, and the new assay was compared to the OIE-accepted conventional PCR test and virus isolation in cell culture. The IHNV N gene RT-rPCR had 100% DSp and DSe and a higher estimated diagnostic odds ratio (DOR) than virus culture or conventional PCR. The RT-rPCR assay was highly repeatable within a laboratory and highly reproducible between laboratories. Field testing of the assay was conducted on a random sample of juvenile steelhead collected from a hatchery raceway experiencing an IHN epizootic. The RT-rPCR detected a greater number of positive samples than cell culture and there was 40% agreement between the 2 tests. Overall, the RT-rPCR assay was highly sensitive, specific, repeatable and reproducible and is suitable for use in a diagnostic setting.

  16. Comparison of PCR, Wright agglutination test and blood culture for diagnosis of brucellosis in suspected patients.

    Science.gov (United States)

    Hekmatimoghaddam, Seyedhosssein; Sadeh, Maryam; Khalili, Mohammad Bagher; Mollaabedin, Mansour; Sazmand, Alireza

    2013-11-15

    Brucellosis has long been prevalent in Iran, with considerable medical and economic importance. Timely diagnosis is needed for early management and effective prevention of its consequences in human beings and animals. Current diagnostic methods impose peculiar challenges in terms of analytical method performance. This study compares diagnostic sensitivity, specificity, predictive Value of Positive (PVP) and Predictive Value of Negative (PVN) for Polymerase Chain Reaction (PCR), Wright agglutination test and blood culture used for patients suspected of brucellosis. In 120 patients clinically suspected of brucellosis and referred by physicians to the Yazd central Medical Laboratory, some relevant demographic, occupational, nutritional and clinical data were collected. Also, venous blood samples were drawn for diagnosis of brucellosis using PCR, Wright agglutination test and blood culture techniques. The most frequent symptom of patients was arthralgia (82 cases, 68.3%). PCR was positive in 25 cases (20.8%), wright test in 21 patients (17.5%) and blood culture in 6 cases (5%). In 20 out of 21 wright-positive cases, PCR was positive and all of the culture-positive patients had positive PCR. Sensitivity, specificity, PVP and PVN of blood culture compared to PCR (as the gold standard test) were 24, 100, 100 and 86%, respectively, but the above parameters when PCR is compared with blood culture (as gold standard) were 100, 83, 24 and 95%, respectively. PCR has better analytical performances than blood culture for diagnosis of brucellosis and is suitable for confirmation of Wright-positive cases.

  17. Study of the bacterial diversity of foods: PCR-DGGE versus LH-PCR.

    Science.gov (United States)

    Garofalo, Cristiana; Bancalari, Elena; Milanović, Vesna; Cardinali, Federica; Osimani, Andrea; Sardaro, Maria Luisa Savo; Bottari, Benedetta; Bernini, Valentina; Aquilanti, Lucia; Clementi, Francesca; Neviani, Erasmo; Gatti, Monica

    2017-02-02

    The present study compared two culture-independent methods, polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) and length-heterogeneity polymerase chain reaction (LH-PCR), for their ability to reveal food bacterial microbiota. Total microbial DNA and RNA were extracted directly from fourteen fermented and unfermented foods, and domain A of the variable regions V1 and V2 of the 16S rRNA gene was analyzed through LH-PCR and PCR-DGGE. Finally, the outline of these analyses was compared with bacterial viable counts obtained after bacterial growth on suitable selective media. For the majority of the samples, RNA-based PCR-DGGE revealed species that the DNA-based PCR-DGGE was not able to highlight. When analyzing either DNA or RNA, LH-PCR identified several lactic acid bacteria (LAB) and coagulase negative cocci (CCN) species that were not identified by PCR-DGGE. This phenomenon was particularly evident in food samples with viable loadsPCR was able to detect a higher number of peaks in the analyzed food matrices relative to species identified by PCR-DGGE. In light of these findings, it may be suggested that LH-PCR shows greater sensitivity than PCR-DGGE. However, PCR-DGGE detected some other species (LAB included) that were not detected by LH-PCR. Therefore, certain LH-PCR peaks not attributed to known species within the LH-PCR database could be solved by comparing them with species identified by PCR-DGGE. Overall, this study also showed that LH-PCR is a promising method for use in the food microbiology field, indicating the necessity to expand the LH-PCR database, which is based, up to now, mainly on LAB isolates from dairy products.

  18. Our love for animals.

    Science.gov (United States)

    Scruton, Roger

    2013-12-01

    Love does not necessarily benefit its object, and cost-free love may damage both object and subject. Our love of animals mobilises several distinct human concerns and should not be considered always as a virtue or always as a benefit to the animals themselves. We need to place this love in its full psychological, cultural, and moral context in order to assess what form it ought to take if animals are to benefit from it.

  19. PRINCIPLES OF ANIMAL BREEDING

    OpenAIRE

    2014-01-01

    University textbook Principles of Animal Breeding is intended for students of agriculture and veterinary medicine. The material is the adapted curricula of undergraduate and graduate level studies in the framework of which the modules Principles of animal breeding as well as Basics of genetics and selection of animals attended are listened. The textbook contains 14 chapters and a glossary of terms. Its concept enables combining fundamental and modern knowledge in the ...

  20. Algorithm Animation with Galant.

    Science.gov (United States)

    Stallmann, Matthias F

    2017-01-01

    Although surveys suggest positive student attitudes toward the use of algorithm animations, it is not clear that they improve learning outcomes. The Graph Algorithm Animation Tool, or Galant, challenges and motivates students to engage more deeply with algorithm concepts, without distracting them with programming language details or GUIs. Even though Galant is specifically designed for graph algorithms, it has also been used to animate other algorithms, most notably sorting algorithms.

  1. The development of a real-time PCR to detect pathogenic Leptospira species in kidney tissue.

    Science.gov (United States)

    Fearnley, C; Wakeley, P R; Gallego-Beltran, J; Dalley, C; Williamson, S; Gaudie, C; Woodward, M J

    2008-08-01

    A LightCycler real-time PCR hybridization probe-based assay that detects a conserved region of the16S rRNA gene of pathogenic but not saprophytic Leptospira species was developed for the rapid detection of pathogenic leptospires directly from processed tissue samples. In addition, a differential PCR specific for saprophytic leptospires and a control PCR targeting the porcine beta-actin gene were developed. To assess the suitability of these PCR methods for diagnosis, a trial was performed on kidneys taken from adult pigs with evidence of leptospiral infection, primarily a history of reproductive disease and serological evidence of exposure to pathogenic leptospires (n=180) and aborted pig foetuses (n=24). Leptospire DNA was detected by the 'pathogenic' specific PCR in 25 tissues (14%) and the control beta-actin PCR was positive in all 204 samples confirming DNA was extracted from all samples. No leptospires were isolated from these samples by culture and no positives were detected with the 'saprophytic' PCR. In a subsidiary experiment, the 'pathogenic' PCR was used to analyse kidney samples from rodents (n=7) collected as part of vermin control in a zoo, with show animals with high microagglutination titres to Leptospira species, and five were positive. Fifteen PCR amplicons from 1 mouse, 2 rat and 14 pig kidney samples, were selected at random from positive PCRs (n=30) and sequenced. Sequence data indicated L. interrogans DNA in the pig and rat samples and L. inadai DNA, which is considered of intermediate pathogenicity, in the mouse sample. The only successful culture was from this mouse kidney and the isolate was confirmed to be L. inadai by classical serology. These data suggest this suite of PCRs is suitable for testing for the presence of pathogenic leptospires in pig herds where abortions and infertility occur and potentially in other animals such as rodents.

  2. Recuperación de boro mediante extracción y procesos de membrana

    OpenAIRE

    Gallego Latorre, Maria Irene

    2008-01-01

    En este proyecto se ha estudiado la posibilidad de extraer boro en forma de ácido bórico mediante extractantes neutros. En primer lugar se han realizado los experimentos de extracción líquido-líquido y después se han realizado pruebas de extracción mediante membranas planas soportadas. Las pruebas de extracción líquido-líquido han permitido estudiar el comportamiento del boro ante diferentes extractantes neutros (2,2,4-Trimethyl-1,3-pentanodiol , 2- Butyl-2-ethyl-1,3-p...

  3. Modulación del crecimiento vertebral mediante electrocoagulación hemicircunferencial vertebral asistida

    OpenAIRE

    2009-01-01

    Nuestro trabajo está basado en la posibilidad de controlar el desarrollo asimétrico de los cartílagos de crecimiento vertebral, mediante la realización de una fisiodesis hemivertebral, con electrocoagulación, videoasistida por toracoscópica. Se realizará en cinco niveles torácicos, con un abordaje anterior mínimamente invasivo. Por lo tanto, planteamos como hipótesis de trabajo que La destrucción de las fisis de crecimiento vertebral mediante electrocoagulación, videoasistida por vía toracosc...

  4. Control de un péndulo invertido mediante nodos CAN

    OpenAIRE

    2004-01-01

    En líneas generales, el propósito de este proyecto, es realizar el control de la maqueta de un péndulo invertido del departamento de ESAII. Esta maqueta está compuesta por una estructura de aluminio, sobre la cual un carro se desliza sobre una superficie plana enganchado a un motor mediante una correa. En este motor se encuentra un encoder óptico, en el extremo de su eje mediante un acoplamiento. En el carro se encuentra el péndulo, el cual es doble, es decir el péndulo cuelga por delante...

  5. Control PID de un secador mediante autómatas programables conectados por ethernet

    OpenAIRE

    2012-01-01

    Este proyecto final de carrera se centra en el control PID de temperatura de un secador convencional mediante autómatas programables conectados por Ethernet. Se busca con ello la configuración de un red de comunicaciones entre dos PLC’s industriales (Programmable Logic Controller en sus siglas en inglés) y el control de temperatura mediante un controlador PID. Los controladores lógicos programables o PLC’s son dispositivos electrónicos muy usados en automatización industrial...

  6. Tratamiento de tumores óseos benignos mediante legrado y relleno con metilmetacrilato

    OpenAIRE

    Orduña Serra, Modesto; Fernández Sabaté, Alfons; Amorós, P.

    1989-01-01

    Se expone la experiencia en ocho lesiones óseas tumorales y paratumorales benignas tratadas mediante legrado de la cavidad y posterior relleno mediante metilmetacrilato con contraste baritado. El seguimiento de los casos entre año y medio y once años permite que nos afirmemos en la bondad de los resultados en cuanto a función articular resistencia de la extremidad y control del tumor. Comentamos las estadísticas publicadas en las que destacan los casos de tumor de células gigantes (66 por 100...

  7. Daño muscular inducido mediante ejercicio y sus efectos en el rendimiento

    OpenAIRE

    Valenzuela Tallón, Pedro Luis

    2014-01-01

    El presente trabajo pretende introducir un tema de gran controversia: el daño muscular inducido mediante ejercicio (DMIE). Se ha realizado una revisión bibliográfica de la literatura científica referente al DMIE para exponer la teoría más actual en cuanto al mecanismo que lo provoca y los cambios biológicos y funcionales producidos tras éste. El DMIE comienza como un proceso mecánico mediante el cual se dañan las estructuras de las fibras musculares, lo cual da lugar a un proceso inflamatorio...

  8. 3D Animation Essentials

    CERN Document Server

    Beane, Andy

    2012-01-01

    The essential fundamentals of 3D animation for aspiring 3D artists 3D is everywhere--video games, movie and television special effects, mobile devices, etc. Many aspiring artists and animators have grown up with 3D and computers, and naturally gravitate to this field as their area of interest. Bringing a blend of studio and classroom experience to offer you thorough coverage of the 3D animation industry, this must-have book shows you what it takes to create compelling and realistic 3D imagery. Serves as the first step to understanding the language of 3D and computer graphics (CG)Covers 3D anim

  9. Animal MRI Core

    Data.gov (United States)

    Federal Laboratory Consortium — The Animal Magnetic Resonance Imaging (MRI) Core develops and optimizes MRI methods for cardiovascular imaging of mice and rats. The Core provides imaging expertise,...

  10. Application of Reverse Transcription-PCR and Real-Time PCR in Nanotoxicity Research

    Science.gov (United States)

    Mo, Yiqun; Wan, Rong; Zhang, Qunwei

    2016-01-01

    Reverse transcription-polymerase chain reaction (RT-PCR) is a relatively simple and inexpensive technique to determine the expression level of target genes and is widely used in biomedical science research including nanotoxicology studies for semiquantitative analysis. Real-time PCR allows for the detection of PCR amplification in the exponential growth phase of the reaction and is much more quantitative than traditional RT-PCR. Although a number of kits and reagents for RT-PCR and real-time PCR are commercially available, the basic principles are the same. Here, we describe the procedures for total RNA isolation by using TRI Reagent, for reverse transcription (RT) by M-MLV reverse transcriptase, and for PCR by GoTaq® DNA Polymerase. And real-time PCR will be performed on an iQ5 multicolor real-time PCR detection system by using iQ™ SYBR Green Supermix. PMID:22975959

  11. DETERMINACIÓN DE SEXO EN AVES MEDIANTE HERRAMIENTAS MOLECULARES

    Directory of Open Access Journals (Sweden)

    MATTA CAMACHO NUBIA E.

    2009-04-01

    Full Text Available

    RESUMEN

    La ausencia de dimorfismo sexual en los estadios juveniles y durante la edad adulta de gran cantidad de especies de aves, dificulta o imposibilita la determinación del sexo basados en el fenotipo. El empleo de marcadores moleculares para determinar el sexo de las aves es una herramienta útil debido a la exactitud y rapidez de los resultados y a su vez se constituye en un método que minimiza el estrés durante la toma de muestra, comparado con otras técnicas invasivas que pudieran afectar la salud o estabilidad biológica del animal. La determinación temprana del sexo en aves resulta de especial relevancia cuando se consideran programas de conservación ex situ, producción, explotación y estudios de ecología de poblaciones. Esta revisión presenta las metodologías usadas para determinar el sexo, haciendo especial énfasis en herramientas moleculares, presentando sus ventajas y limitaciones.

    Palabras clave: dimorfismo sexual, aves, CHD, tipificación molecular cromosoma W, cromosoma Z.


    ABSTRACT

    The lack of sexual dimorphism in nestling, juvenile or adult birds of large number of avian species, makes it difficult or impossible sex determination based on phenotipic characteristics. To use molecular markers for bird sex determination is a rapid and safe procedure; moreover this methodology minimizes the stress during sampling, compared to other invasive techniques that could affect the health or biological stability of the animal. The early sex determination in birds is of particular importance when considering ex situ conservation programs, production

  12. Epidemiological studies on animal and human trichinellosis in Estonia

    Directory of Open Access Journals (Sweden)

    Järvis T.

    2001-06-01

    Full Text Available From 1992 to 1999, muscle samples from 814 sylvatic animals and 1,173 domestic and synanthropic animals were collected in 15 districts of Estonia ; the prevalence of trichinellosis ranged from 1.0 % to 79.4 % for sylvatic animals and from 0.6 % to 24.5 % for domestic or synanthropic animals and for animals from fur-bearing farms. The most important reservoirs of Trichinella in nature were the raccoon dog, the red fox, the lynx and the wolf. Three species of Trichinella (T. spiralis, T. nativa, and T. britovi were identified by several types of PCR-based analyses. Meat from sylvatic animals was the main source of Trichinella infection for humans.

  13. The polymerase chain reaction (PCR): general methods.

    Science.gov (United States)

    Waters, Daniel L E; Shapter, Frances M

    2014-01-01

    The polymerase chain reaction (PCR) converts very low quantities of DNA into very high quantities and is the foundation of many specialized techniques of molecular biology. PCR utilizes components of the cellular machinery of mitotic cell division in vitro which respond predictably to user inputs. This chapter introduces the principles of PCR and discusses practical considerations from target sequence definition through to optimization and application.

  14. Detección sensible y específica de Mycobacterium tuberculosis a partir de muestras clínicas, mediante la amplificación de un elemento repetitivo de la familia REP13E12

    OpenAIRE

    Christian Baldeviano V; Carmen Luna C; Tatiana Cáceres N; Roger Calderón E

    2007-01-01

    Objetivos: Diseñar e implementar un método de PCR para la detección sensible y específica de M. tuberculosis, orientado al oportuno diagnóstico de la tuberculosis mediante su adecuada aplicación en muestras clínicas. Materiales y métodos: Se diseñaron oligonucleótidos específicos para la amplificación de un fragmento de 318pb luego de una alineación múltiple de secuencias REP13E12 de M. tuberculosis. Se realizó la estandarización del método de PCR y se evaluó la sensibilidad y especificidad d...

  15. Genotyping of the Holstein-Friesian crossbred cattle for CD18 gene using PCR-RFLP

    Directory of Open Access Journals (Sweden)

    A. S. Khade

    2014-05-01

    Full Text Available Aim: The present study was undertaken in Holstein-Friesian (HF crossbred cattle with the objective to find out genotype of HF crossbred cattle for Bovine Leucocyte Adhesion Deficiency (BLAD by using PCR-RFLP. Materials and Methods: 50 blood samples were collected from HF crossbred cattle and subjected to PCR. The amplified PCR products were digested using Taq I restriction enzyme at 65 oC overnight. After restriction digestion, the final PCR products were electrophoresed on 2.5 % agarose gel. Results: All the 50 animals under present investigation were found to be normal as the amplified PCR product upon digestion with Taq I restriction enzyme, revealed two bands of 313 bp and 54 bp for normal animals. Conclusions: In the present investigation D128G carrier frequency was found to be 0 %. However, recent reports suggest that the mutant gene has already been observed in the HF crossbred cattle population of India, which makes it necessary to screen the animals to avoid the risk of spreading BLAD in the breeding cattle population.

  16. Identification of Chlamydial species in crocodiles and chickens by PCR-HRM curve analysis.

    Science.gov (United States)

    Robertson, T; Bibby, S; O'Rourke, D; Belfiore, T; Agnew-Crumpton, R; Noormohammadi, A H

    2010-10-26

    Recently, a PCR protocol (16SG), targeting 16S rRNA gene coupled with high resolution melt (HRM) curve analysis was developed in our laboratory and shown to reliably detect and identify the seven different Chlamydiaceae spp. In this study, the potential of this method was assessed for detection and differentiation of Chlamydiosis in clinical specimens. Of the total number of 733 specimens from a range of animal species, 219 (30%) were found positive by 16SG PCR. When a sufficient amount of DNA was available (64 submissions), amplicons generated by the 16SG PCR were subjected to HRM curve analysis and results were compared to that of nucleotide sequencing. In all instances, the infecting Chlamydiaceae spp. was genotyped according to the identity of its nucleotide sequence to a reference species. Analysis of the HRM curves and nucleotide sequences from 16SG PCR amplicons also revealed the occurrence of a Chlamydophila-like, a Parachlamydia-like and a variant of Chlamydophila psittaci in chickens. These results reveal the potential of 16SG PCR-HRM curve analysis for rapid and simultaneous detection and identification of Chlamydiaceae spp. in animals and demonstrate the capacity of this system for rapid identification of new Chlamydiaceae spp. in animals during routine diagnostic testings.

  17. DETERMINACIÓN DE SEXO EN AVES MEDIANTE HERRAMIENTAS MOLECULARES

    Directory of Open Access Journals (Sweden)

    NUBIA E. MATTA CAMACHO

    2009-01-01

    Full Text Available La ausencia de dimorfismo sexual en los estadios juveniles y durante la edad adulta de gran cantidad de especies de aves, dificulta o imposibilita la determinación del sexo basados en el fenotipo. El empleo de marcadores moleculares para determinar el sexo de las aves es una herramienta útil debido a la exactitud y rapidez de los resultados y a su vez se constituye en un método que minimiza el estrés durante la toma de muestra, comparado con otras técnicas invasivas que pudieran afectar la salud o estabilidad biológica del animal. La determinación temprana del sexo en aves resulta de especial relevancia cuando se consideran programas de conservación ex situ, producción, explotación y estudios de ecología de poblaciones. Esta revisión presenta las metodologías usadas para determinar el sexo, haciendo especial énfasis en herramientas moleculares, presentando sus ventajas y limitaciones.

  18. Restriction endonucleases digesting DNA in PCR buffer

    Institute of Scientific and Technical Information of China (English)

    LIU Xue-dong; ZHENG Dong; ZHOU Yan-na; MAO Wei-wei; MA Jian-zhang

    2005-01-01

    Six commonly used restriction endonucleases (Res) (Acc I, Ban II, EcoR I, Hind III, Sac I, Sca I) were tested for their ability to directly digest DNA completely in the Polymerase Chain Reaction (PCR) buffers. The results showed that: with the requirement for additional magnesium supplemented as activator, Res, except EcoR I appeared star activity, completely digested unmethylated lambda DNA after overnight incubation in PCR buffer and functioned as equally well as in recommended Restriction Enzyme Buffer provided with each enzyme; all Res tested completely digested PCR products in PCR buffer, it implied digestion of PCR products may often be performed directly in the PCR tube without the requirement for any precipitation or purification steps; and the concentration of MgCl2 from 2.5 mmol·L-1 to 10 mmol·L-1 did not significantly affect activity of Res in PCR buffer. This simplified method for RE digestion of PCR products could have applications in restriction fragment length polymorphism (RFLP) analysis and single-stranded conformational polymorphism (SSCP) analysis of large PCR products. However, usage of this procedure for cloning applications needs further data.

  19. Detection of Eperythrozoon wenyoni by PCR assay

    Institute of Scientific and Technical Information of China (English)

    Jian WANG; Yutao ZHU; Jianhua QIN; Fumei ZHANG; Yuelan ZHAO

    2009-01-01

    The objective of this research was to develop a detection method for Eperythrozoon wenyoni infection using polymerase chain reaction (PCR) assay technique. A pair of primers was designed and synthesized according to the conservative sequence 16S rRNA. The PCR assay was performed with the primers. A 985-bp fragment was amplified by using PCR. The amplified fragments with the expected size were identified by EcoR I restriction digestion. The crossing-reaction, specific-reaction and duplicate-reaction indicated that the PCR method is a specific, sensitive, fast and effective method for diagnosing E. Wenyoni infection at group level.

  20. Real-time PCR in microfluidic devices

    Science.gov (United States)

    Becker, Holger; Hlawatsch, Nadine; Klemm, Richard; Moche, Christian; Hansen-Hagge, Thomas; Gärtner, Claudia

    2014-03-01

    A central method in a standard biochemical laboratory is represented by the polymerase chain reaction (PCR), therefore many attempts have been performed so far to implement this technique in lab-on-a-chip (LOC) devices. PCR is an ideal candidate for miniaturization because of a reduction of assay time and decreased costs for expensive bio-chemicals. In case of the "classical" PCR, detection is done by identification of DNA fragments electrophoretically separated in agarose gels. This method is meanwhile frequently replaced by the so-called Real-Time-PCR because here the exponential increase of amplificates can be observed directly by measurement of DNA interacting fluorescent dyes. Two main methods for on-chip PCRs are available: traditional "batch" PCR in chambers on a chip using thermal cycling, requiring about 30 minutes for a typical PCR protocol and continuous-flow PCR, where the liquid is guided over stationary temperature zones. In the latter case, the PCR protocol can be as fast as 5 minutes. In the presented work, a proof of concept is demonstrated for a real-time-detection of PCR products in microfluidic systems.

  1. Pitfalls in PCR troubleshooting: Expect the unexpected?

    Science.gov (United States)

    Schrick, Livia; Nitsche, Andreas

    2016-01-01

    PCR is a well-understood and established laboratory technique often used in molecular diagnostics. Huge experience has been accumulated over the last years regarding the design of PCR assays and their set-up, including in-depth troubleshooting to obtain the optimal PCR assay for each purpose. Here we report a PCR troubleshooting that came up with a surprising result never observed before. With this report we hope to sensitize the reader to this peculiar problem and to save troubleshooting efforts in similar situations, especially in time-critical and ambitious diagnostic settings.

  2. Pitfalls in PCR troubleshooting: Expect the unexpected?

    Directory of Open Access Journals (Sweden)

    Livia Schrick

    2016-01-01

    Full Text Available PCR is a well-understood and established laboratory technique often used in molecular diagnostics. Huge experience has been accumulated over the last years regarding the design of PCR assays and their set-up, including in-depth troubleshooting to obtain the optimal PCR assay for each purpose. Here we report a PCR troubleshooting that came up with a surprising result never observed before. With this report we hope to sensitize the reader to this peculiar problem and to save troubleshooting efforts in similar situations, especially in time-critical and ambitious diagnostic settings.

  3. Companion Animals. [Information Packet.

    Science.gov (United States)

    National Anti-Vivisection Society, Chicago, IL.

    This collection of articles reprinted from other National Anti-Vivisection Society (NAVS) publications was compiled to educate the public on issues of importance to NAVS concerning companion animals. Topics covered include spaying and neutering, animal safety, pet theft, and the use of cats and dogs in research. The article on spaying and…

  4. Animals as disgust elicitors

    DEFF Research Database (Denmark)

    Kasperbauer, Tyler Joshua

    2015-01-01

    This paper attempts to explain how and why nonhuman animals elicit disgust in human beings. I argue that animals elicit disgust in two ways. One is by triggering disease–protection mechanisms, and the other is by eliciting mortality salience, or thoughts of death. I discuss how these two types of...

  5. Indian draught animals power

    Directory of Open Access Journals (Sweden)

    K. L. Phaniraja

    Full Text Available With the modernization of agriculture, the use of mechanical power in agriculture has increased but draught animal power (DAP continues to be used on Indian farms due to small holdings and hill agriculture. More than 55% of the total cultivated area is still being managed by using draught animals as against about 20% by tractors. India possessed the finest breeds of draught animals. Bullocks, buffaloes and camels are the major draught animals for field operations. Horses, mules, donkeys, yak and mithun are the pack animals for transport. The quality of work from the draught animals depends upon the power developed by them. The design of traditional implements is based on long experience and these have served the purpose of the farmers. However there is plenty of scope to improve the design based on animal-machine-environment interaction so as to have more output and increased efficiency without jeopardizing animal health. [Vet World 2009; 2(10.000: 404-407

  6. Designing for animals

    NARCIS (Netherlands)

    Bouwhuis, T.

    2012-01-01

    This "designers' manual" is made during the TIDO-course AR0531 Smart & Bioclimatic Design. Providing living space for animals in cities is an underexposed subject in the practice of urban designers. We encounter the results of conflicting situations between humans and animals almost every day, and

  7. Hazardous marine animals.

    Science.gov (United States)

    Auerbach, P S

    1984-08-01

    Both traumatic injury and the damage inflicted by envenomating marine animals are considered in this article. Among the creatures causing traumatic injury are sharks, barracudas, moray eels, and needlefish. Envenomating animals include sponges, coelenterates, coral, various mollusks, sea urchins, sea cucumbers, stingrays, sea snakes, and others.

  8. Animals in the Classroom

    Science.gov (United States)

    Roy, Ken

    2011-01-01

    Use of animals in middle school science classrooms is a curriculum component worthy of consideration, providing proper investigation and planning are addressed. A responsible approach to this action, including safety, must be adopted for success. In this month's column, the author provides some suggestions on incorporating animals into the…

  9. Political Communication with Animals

    NARCIS (Netherlands)

    E. Meijer

    2013-01-01

    In this article I sketch the outlines of a theory of political human-animal conversations, based on ideas about language that I borrow from Ludwig Wittgenstein’s later work, in particular his notion of language-games. I present this theory as a supplement to the political theory of animal rights Sue

  10. The Classroom Animal: Snails.

    Science.gov (United States)

    Kramer, David S.

    1985-01-01

    Points out that snails are interesting and easily-managed classroom animals. One advantage of this animal is that it requires no special attention over weekends or holidays. Background information, anatomy, reproduction, and feeding are discussed, along with suggestions for housing aquatic and/or land snails. (DH)

  11. Animal Care Use Committees.

    Science.gov (United States)

    Snyder, Margaret D.; And Others

    1992-01-01

    Describes the structure, activities, responsibilities, and practices of animal care and use committees established to review classroom activities and student research using animals. Provides six hypothetical situations with suggested solutions to test a committee's decision-making ability. Includes a proposed activity form for teachers. (MDH)

  12. Small Animal Care.

    Science.gov (United States)

    Livesey, Dennis W.; Fong, Stephen

    This small animal care course guide is designed for students who will be seeking employment in veterinary hospitals, kennels, grooming shops, pet shops, and small-animal laboratories. The guide begins with an introductory section that gives the educational philosophy of the course, job categories and opportunities, units of instruction required…

  13. Animal ethics dilemma

    DEFF Research Database (Denmark)

    Dich, Trine; Hansen, Tina; Algers, Anne

    2006-01-01

    'Animal Ethics Dilemma' is a freely available computer-supported learning tool (www.animalethicsdilemma.net or www.aedilemma.net) which has been developed primarily for veterinary undergraduates but is applicable also to students in other fields of animal science. The objectives of the computer...... program are to promote students' understanding of the ethics related to animal use, to illustrate ethical dilemmas that arise in animal use, to broaden students' moral imagination, and to enable students to differentiate between types of ethical argument. The program comprises five case studies: (1......) the blind hens; (2) ANDi the genetically modified monkey; (3) euthanasia of a healthy dog; (4) animal slaughter; and (5) rehabilitation of seals. Special consideration has been given to enhancing the pedagogic value of the program. Students can control their learning by selecting a variety of ways...

  14. Workshop on molecular animation.

    Science.gov (United States)

    Bromberg, Sarina; Chiu, Wah; Ferrin, Thomas E

    2010-10-13

    From February 25 to 26, 2010, in San Francisco, the Resource for Biocomputing, Visualization, and Informatics (RBVI) and the National Center for Macromolecular Imaging (NCMI) hosted a molecular animation workshop for 21 structural biologists, molecular animators, and creators of molecular visualization software. Molecular animation aims to visualize scientific understanding of biomolecular processes and structures. The primary goal of the workshop was to identify the necessary tools for producing high-quality molecular animations, understanding complex molecular and cellular structures, creating publication supplementary materials and conference presentations, and teaching science to students and the public. Another use of molecular animation emerged in the workshop: helping to focus scientific inquiry about the motions of molecules and enhancing informal communication within and between laboratories.

  15. Sketching with animation

    DEFF Research Database (Denmark)

    Vistisen, Peter

    , the aim is to present a range of analytical arguments and experimental results that indicate the need for a systematic approach to realising the potential of animation within design sketching. This will establish the foundation for what we label animation-based sketching.......This book offers a contribution to the theory, method and techniques involved in the use of animation as a tool for temporal design sketching. Lifted from its traditional role as a genre of entertainment and art and reframed in the design domain, animation offers support during the early phases...... of exploring and assessing the potential of new and emerging digital technologies. This approach is relatively new and has been touched upon by few academic contributions in the past. Thus, the aim of the text is not to promote a claim that sketching with animation is an inherently new phenomenon. Instead...

  16. Is animal experimentation fundamental?

    Science.gov (United States)

    d'Acampora, Armando José; Rossi, Lucas Félix; Ely, Jorge Bins; de Vasconcellos, Zulmar Acciolli

    2009-01-01

    The understanding about the utilization of experimental animals in scientific research and in teaching is many times a complex issue. Special attention needs to be paid to attain the understanding by the general public of the importance of animal experimentation in experimental research and in undergraduate medical teaching. Experimental teaching and research based on the availability of animals for experimentation is important and necessary for the personal and scientific development of the physician-to-be. The technological arsenal which intends to mimic experimentation animals and thus fully replace their use many times does not prove to be compatible with the reality of the living animal. The purpose of this paper is to discuss aspects concerning this topic, bringing up an issue which is complex and likely to arouse in-depth reflections.

  17. Cupper in animal tissues

    Directory of Open Access Journals (Sweden)

    Maximino Huerta Bravo

    2010-12-01

    Full Text Available Cupper is an essential element for plants, animals and humans. Under certain circumstances, cupper excessive consumption could result in animal and human intoxication. In order to ensure safe and innocuous and safe foods for Mexicans, government create legislation as Norma Oficial Mexicana to establish the maximum levels of residues, particularly cupper in liver, kidney and muscle of human consumption animals. Liver in Mexico ruminant animals regularly contain 60 mg Cu/kg, which is the legal limit for this metal. This demands a review of the actual legislation. The strict application of this Norma will limit the commercialization of these viscera, since approximately 50% will exceed the legal limit for cupper. A potential hazard for human health, especially young people, is found in the constant ovine liver consumption feed with animal excretes with higher amount of supplementary cupper.

  18. Towards an animated JPEG

    Science.gov (United States)

    Theytaz, Joël.; Yuan, Lin; McNally, David; Ebrahimi, Touradj

    2016-09-01

    Recently, short animated image sequences have become very popular in social networks. Most animated images are represented in GIF format. In this paper we propose an animated JPEG format, called aJPEG, which allows the standard JPEG format to be extended in a backward compatible way in order to cope with animated images. After presenting the proposed format, we illustrate it using two prototype applications: the first in form of a GIF-to-aJPEG converter on a personal computer and the second in form of an aJPEG viewer on a smart phone. The paper also reports the performance evaluation of aJPEG when compared to GIF. Experimental results show that aJPEG outperforms animated GIF in both file size overhead and image quality.

  19. Discrimination of Shark species by simple PCR of 5S rDNA repeats

    Directory of Open Access Journals (Sweden)

    Danillo Pinhal

    2008-01-01

    Full Text Available Sharks are suffering from intensive exploitation by worldwide fisheries leading to a severe decline in several populations in the last decades. The lack of biological data on a species-specific basis, associated with a k-strategist life history make it difficult to correctly manage and conserve these animals. The aim of the present study was to develop a DNA-based procedure to discriminate shark species by means of a rapid, low cost and easily applicable PCR analysis based on 5S rDNA repeat units amplification, in order to contribute conservation management of these animals. The generated agarose electrophoresis band patterns allowed to unequivocally distinguish eight shark species. The data showed for the first time that a simple PCR is able to discriminate elasmobranch species. The described 5S rDNA PCR approach generated species-specific genetic markers that should find broad application in fishery management and trade of sharks and their subproducts.

  20. Survey for Toxoplasma gondii by PCR detection in meat for human consumption in Colombia.

    Science.gov (United States)

    Franco-Hernandez, Erika N; Acosta, Alejandro; Cortés-Vecino, Jesús; Gómez-Marín, Jorge Enrique

    2016-02-01

    The overall risk for toxoplasmosis in meat produced in Colombia is unknown. We analyzed by PCR assay meat samples for human consumption in two types of plants in Colombia: 120 samples from class I plants (60 samples from chicken, 30 from swine and 30 from beef) and 60 from class II plants (30 samples from beef and 30 from swine). Presence of Toxoplasma DNA was established by targeted B1 nested PCR assay. We detected 79 (43%) samples that were positive by B1 nested PCR (33 from chicken, 22 from beef, and 24 from pork). No differences were found by region or species. Eleven positive samples were confirmed by sequencing of the B1 repeated region. Some polymorphisms were detected without relation with clonal groups nor meat species. Food animals are highly exposed to Toxoplasma in Colombia. Detailed studies are needed to establish the reasons for differences in Toxoplasma prevalence between farms, regarding practices of animal food production.

  1. Animal Diseases and Your Health

    Science.gov (United States)

    Animal diseases that people can catch are called zoonoses. Many diseases affecting humans can be traced to animals or animal products. You can get a disease directly from an animal, or indirectly, through the ...

  2. Evaluation of AMPLICOR Neisseria gonorrhoeae PCR using cppB nested PCR and 16S rRNA PCR.

    Science.gov (United States)

    Farrell, D J

    1999-02-01

    Certain strains of Neisseria subflava and Neisseria cinerea are known to produce false-positive results with the AMPLICOR Neisseria gonorrhoeae PCR (Roche Diagnostic Systems, Branchburg, N.J.). The analytical sensitivity and analytical specificity of three PCR tests were assessed with 3 geographically diverse N. gonorrhoeae strains and 30 non-N. gonorrhoeae Neisseria spp. The sensitivities of the in-house nested cppB gene and the 16S rRNA PCR methods were greater than that of the AMPLICOR N. gonorrhoeae PCR with purified DNA from all 3 N. gonorrhoeae strains. Six of 14 clinical strains of N. subflava (1 from a vaginal swab, 5 from respiratory sites) produced false-positive AMPLICOR N. gonorrhoeae PCR results and were negative by the two other PCR methods. When applied to 207 clinical specimens selected from a population with a high prevalence ( approximately 9%) of infection, the results for 15 of 96 (15.6%) AMPLICOR-positive specimens and 14 of 17 (82.3%) AMPLICOR-equivocal specimens were not confirmed by the more sensitive nested cppB PCR method. Only 2 of 94 (2.1%) of AMPLICOR N. gonorrhoeae PCR-negative specimens from the same population tested positive by the nested cppB method. These results suggest that for this population the AMPLICOR N. gonorrhoeae PCR test is suitable as a screening test only and all positive results should be confirmed by a PCR method that is more specific and at least as sensitive. This study also illustrates that caution should be used when introducing commercially available nucleic acid amplification-based diagnostic tests into the regimens of tests used for populations not previously tested with these products.

  3. Sex determination in 6 bovid species by duplex PCR.

    Science.gov (United States)

    Prashant; Gour, Digpal S; Dubey, Prem P; Jain, Anubhav; Gupta, Subhash C; Joshi, Balwinder K; Kumar, Dinesh

    2008-01-01

    Sex determination in domestic animals is of potential value to livestock breeding programs. The aim of this study was to develop a simple and accurate PCR-based sex determination protocol, which can be applicable to 6 major domesticated species of the family Bovidae, viz. Bos frontalis, B. grunniens, B. indicus, Bubalus bubalis, Capra hircus, and Ovis aries. In silico analysis was done to identify conserved DNA sequence in the HMG box region of the sex-determining region of the Y-chromosome (SRY gene) across the bovids. Duplex PCR assay, including the SRY gene and the GAPDH housekeeping gene, was optimized by using genomic DNA extracted from blood samples of known sex. It was possible to identify the sex of animals by amplifying both gender-specific (SRY) and autosomal (GAPDH) genes simultaneously in the duplex reaction, with the male yielding two bands and the female one band. The protocol was subjected to a blind test that showed a 100 percent specificity and accuracy, thus it can be used in sex determination in livestock breeding programs.

  4. Método de eliminación de trihalometanos y/o contaminantes emergentes mediante plasma

    OpenAIRE

    Molina, Ricardo; Erra Serrabasa, Pilar; Bertrán, Enric; Jover Comas, Eric; Reyes Contreras, Carolina

    2009-01-01

    Método de eliminación de trihalometanos y/o contaminantes emergentes mediante plasma. Se describe un método de eliminación de trihalometanos y contaminantes refractarios en medios acuosos mediante la aplicación directa de plasma para conseguir la degradación de los compuestos contaminantes presentes en el agua.

  5. Antibiotic resistance gene discovery in food-producing animals.

    Science.gov (United States)

    Allen, Heather K

    2014-06-01

    Numerous environmental reservoirs contribute to the widespread antibiotic resistance problem in human pathogens. One environmental reservoir of particular importance is the intestinal bacteria of food-producing animals. In this review I examine recent discoveries of antibiotic resistance genes in agricultural animals. Two types of antibiotic resistance gene discoveries will be discussed: the use of classic microbiological and molecular techniques, such as culturing and PCR, to identify known genes not previously reported in animals; and the application of high-throughput technologies, such as metagenomics, to identify novel genes and gene transfer mechanisms. These discoveries confirm that antibiotics should be limited to prudent uses.

  6. Digital PCR dynamic range is approaching that of real-time quantitative PCR.

    Science.gov (United States)

    Jones, Gerwyn M; Busby, Eloise; Garson, Jeremy A; Grant, Paul R; Nastouli, Eleni; Devonshire, Alison S; Whale, Alexandra S

    2016-12-01

    Digital PCR (dPCR) has been reported to be more precise and sensitive than real-time quantitative PCR (qPCR) in a variety of models and applications. However, in the majority of commercially available dPCR platforms, the dynamic range is dependent on the number of partitions analysed and so is typically limited to four orders of magnitude; reduced compared with the typical seven orders achievable by qPCR. Using two different biological models (HIV DNA analysis and KRAS genotyping), we have demonstrated that the RainDrop Digital PCR System (RainDance Technologies) is capable of performing accurate and precise quantification over six orders of magnitude thereby approaching that achievable by qPCR.

  7. A simplified method to prepare PCR template DNA for screening of transgenic and knockout mice.

    Science.gov (United States)

    Ren, S; Li, M; Cai, H; Hudgins, S; Furth, P A

    2001-03-01

    Polymerase chain reaction (PCR) amplification of DNA is the most widely used technique for screening of large numbers of genetically engineered transgenic or knockout mice (Mus musculus). In this report, we present a new DNA preparation procedure for running diagnostic PCR. In this procedure, mouse ear tissue was used directly for PCR after the tissue underwent brief digestion in a solution containing only proteinase K. Using this method, we have successfully screened several lines of single, double, and triple transgenic and knockout mice. The results are reliable and reproducible. The advantage of this new method is that DNA purification by organic extraction or isolation kit was omitted. DNA purification is the limiting factor in terms of time and money when screening transgenic and knockout mice by PCR. In addition, using ear instead of tail tissue can reduce distress of animals because the samples can be obtained when the mice are labeled by ear punch.

  8. A duplex PCR for the rapid and simultaneous detection of Brucella spp. in human blood samples

    Institute of Scientific and Technical Information of China (English)

    Reza Mirnejad; Mozafar mohamadi; Vahbeh Piranfar; Seied Mojtaba Mortazavi; Reza Kachuei

    2013-01-01

    Objective: To design a duplex PCR for rapid and simultaneous detection of Brucella species. in human blood samples. Methods: Fifty-two peripheral bloods samples were collected from suspicious patients with brucellosis. Following DNA extraction, PCR assay were performed, using three primers that could simultaneously identify and differentiate three major species of pathogenic Brucella in humans and animals. Results: Of the 52 peripheral bloods samples tested, 25 sample (48%) showed positive reactions in PCR. Twelve samples were positive for Brucella abortus (B. abortus) (23%), 13 for Brucella melitensis (B. melitensis) (25%) and 0 for Brucella ovis (B. ovis) (0%). Conclusions: This work de=monstrates that in case where specific primers were utilized, duplex PCR has proved to be a simple, fast, and relatively inexpensive method for simultaneous detection of important species of Brucella in clinical samples.

  9. Canine distemper virus detection by different methods of One-Step RT-qPCR

    Directory of Open Access Journals (Sweden)

    Claudia de Camargo Tozato

    2016-01-01

    Full Text Available ABSTRACT: Three commercial kits of One-Step RT-qPCR were evaluated for the molecular diagnosis of Canine Distemper Virus. Using the kit that showed better performance, two systems of Real-time RT-PCR (RT-qPCR assays were tested and compared for analytical sensitivity to Canine Distemper Virus RNA detection: a One-Step RT-qPCR (system A and a One-Step RT-qPCR combined with NESTED-qPCR (system B. Limits of detection for both systems were determined using a serial dilution of Canine Distemper Virus synthetic RNA or a positive urine sample. In addition, the same urine sample was tested using samples with prior centrifugation or ultracentrifugation. Commercial kits of One-Step RT-qPCR assays detected canine distemper virus RNA in 10 (100% urine samples from symptomatic animals tested. The One-Step RT-qPCR kit that showed better results was used to evaluate the analytical sensitivity of the A and B systems. Limit of detection using synthetic RNA for the system A was 11 RNA copies µL-1 and 110 RNA copies µl-1 for first round System B. The second round of the NESTED-qPCR for System B had a limit of detection of 11 copies µl-1. Relationship between Ct values and RNA concentration was linear. The RNA extracted from the urine dilutions was detected in dilutions of 10-3 and10-2 by System A and B respectively. Urine centrifugation increased the analytical sensitivity of the test and proved to be useful for routine diagnostics. The One-Step RT-qPCR is a fast, sensitive and specific method for canine distemper routine diagnosis and research projects that require sensitive and quantitative methodology.

  10. Validation of RNAi by real time PCR

    DEFF Research Database (Denmark)

    Josefsen, Knud; Lee, Ying Chiu

    2011-01-01

    Real time PCR is the analytic tool of choice for quantification of gene expression, while RNAi is concerned with downregulation of gene expression. Together, they constitute a powerful approach in any loss of function studies of selective genes. We illustrate here the use of real time PCR to veri...

  11. PCR specific for Actinobacillus pleuropneumoniae serotype 3

    DEFF Research Database (Denmark)

    Zhou, L.; Jones, S.C.P.; Angen, Øystein

    2008-01-01

    , but the method has liminations, for example, cross-reactions between serotypes 3, 6, and 8. This study describes the development of a serotype 3-specific PCR, based on the capsule locus, which can be used in a multiplex format with the organism's specific gene apxIV. The PCR test was evaluated on 266 strains...

  12. Testing for Genetically Modified Foods Using PCR

    Science.gov (United States)

    Taylor, Ann; Sajan, Samin

    2005-01-01

    The polymerase chain reaction (PCR) is a Nobel Prize-winning technique that amplifies a specific segment of DNA and is commonly used to test for the presence of genetic modifications. Students use PCR to test corn meal and corn-muffin mixes for the presence of a promoter commonly used in genetically modified foods, the cauliflower mosaic virus 35S…

  13. Digital PCR for detection of citrus pathogens

    Science.gov (United States)

    Citrus trees are often infected with multiple pathogens of economic importance, especially those with insect or mite vectors. Real-time/quantitative PCR (qPCR) has been used for high-throughput detection and relative quantification of pathogens; however, target reference or standards are required. I...

  14. Real-time PCR and PCR-tandem Mass Spectrometry for Biodetection

    Science.gov (United States)

    2005-10-01

    Real - time PCR and PCR- tandem mass spectrometry for biodetection Alvin Fox, University of South Carolina, School of Medicine Report Documentation...TYPE N/A 3. DATES COVERED - 4. TITLE AND SUBTITLE Real - time PCR and PCRtandem mass spectrometry for biodetection 5a. CONTRACT NUMBER 5b...interspace region Bacillus subtilis W23 standard Blank Barn dust House dust Cycle Real - time PCR (16s rRNA) - environmental samples Real - time

  15. Addressing PCR Biases in Environmental Microbiology Studies

    Science.gov (United States)

    Sipos, Rita; Székely, Anna; Révész, Sára; Márialigeti, Károly

    Each step of a molecular environmental microbiology study is prone to errors, though the qualitative and quantitative biases of PCR amplification could result in the most serious biases. One has to be aware of this fact, and well-characterized PCR biases have to be avoided by using target-optimized PCR protocols. The most important tasks are primer and thermal profile optimization. We have shown that primer mismatches, even in the case of universal primers, can cause almost complete missing of common taxa from clone libraries, for example. Similarly high annealing temperatures can drastically distort community composition of the sample in the PCR product. Strategies of primer selection and PCR thermal profile design are discussed in detail.

  16. COLD-PCR: Applications and Advantages.

    Science.gov (United States)

    Zuo, Zhuang; Jabbar, Kausar J

    2016-01-01

    Co-amplification at lower denaturation temperature-based polymerase chain reaction (COLD-PCR) is a single-step amplification method that results in the enhancement of both known and unknown minority alleles during PCR, irrespective of mutation type and position. This method is based on exploitation of the critical temperature, Tc, at which mutation-containing DNA is preferentially melted over wild type. COLD-PCR can be a good strategy for mutation detection in specimens with high nonneoplastic cell content, small specimens in which neoplastic cells are difficult to micro-dissect and therefore enrich, and whenever a mutation is suspected to be present but is undetectable using conventional PCR and sequencing methods. We describe in this chapter our COLD-PCR-based pyrosequencing method for KRAS mutation detection in various clinical samples using DNA extracted from either fresh or fixed paraffin-embedded tissue specimens.

  17. Animal welfare and eggs

    DEFF Research Database (Denmark)

    Andersen, Laura Mørch

    This paper identifies revealed willingness to pay for animal welfare using a panel mixed logit model allowing for correlation between willingness to pay for different types of production. We utilize a unique household level panel, combining real purchases with survey data on perceived public...... and private good attributes of different types of eggs. We find that the estimated correlations are consistent with the levels of animal welfare, and that consumers perceiving a stronger connection between animal welfare and the organic label have higher willingness to pay for organic eggs, even when we...

  18. Animals eponyms in dermatology

    Directory of Open Access Journals (Sweden)

    Nidhi Jindal

    2014-01-01

    Full Text Available The world of Dermatology is flooded with inflexions among clinical conditions and signs and syndromes; making it interesting, but a tougher subject to remember. Signs and syndromes have always fascinated residents, but simultaneously burdened their minds, as these attractive names are difficult to remember. This work was undertaken to review dermatological conditions and signs based on commonly encountered daily words and objects like animals, etc. Fifty dermatological conditions were found to be based on animal eponyms. For example, the usage of animal terminology in dermatology like leonine facies is present in leprosy, sarcoidosis, mycosis fungoides (MF, and airborne contact dermatitis (ABCD.

  19. Environmentally friendly animal litter

    Science.gov (United States)

    Chett, Boxley; McKelvie, Jessica

    2013-08-20

    A method of making an animal litter that includes geopolymerized ash, wherein, the animal litter is made from a quantity of a pozzolanic ash mixed with a sufficient quantity of water and an alkaline activator to initiate a geopolymerization reaction that forms geopolymerized ash. After the geopolymerized ash is formed, it is dried, broken into particulates, and sieved to a desired size. These geopolymerized ash particulates are used to make a non-clumping or clumping animal litter. Odor control may be accomplished with the addition of a urease inhibitor, pH buffer, an odor eliminating agent, and/or fragrance.

  20. Women Protecting Endangered Animals

    Institute of Scientific and Technical Information of China (English)

    1997-01-01

    ON the Yongding River, 40 kilometers south of Beijing lies the Beijing Center for Breeding Endangered Animals.Built more than 10 years ago it is the only rare and endangered animal base in China, incorporating such functions as Scientific research, raising, breeding and medical treatment. There are more than 30 national and international rare species, with a total of more than 1,000 animals. Among them, the snub-nosed golden monkey, Chinese monal pheasant and eared pheasant account for the largest number of man-bred species in the world.

  1. Computer facial animation

    CERN Document Server

    Parke, Frederic I

    2008-01-01

    This comprehensive work provides the fundamentals of computer facial animation and brings into sharper focus techniques that are becoming mainstream in the industry. Over the past decade, since the publication of the first edition, there have been significant developments by academic research groups and in the film and games industries leading to the development of morphable face models, performance driven animation, as well as increasingly detailed lip-synchronization and hair modeling techniques. These topics are described in the context of existing facial animation principles. The second ed

  2. Pre-PCR processing: strategies to generate PCR-compatible samples.

    Science.gov (United States)

    Rådström, Peter; Knutsson, Rickard; Wolffs, Petra; Lövenklev, Maria; Löfström, Charlotta

    2004-02-01

    Polymerase chain reaction (PCR) is recognized as a rapid, sensitive, and specific molecular diagnostic tool for the analysis of nucleic acids. However, the sensitivity and kinetics of diagnostic PCR may be dramatically reduced when applied directly to biological samples, such as blood and feces, owing to PCR-inhibitory components. As a result, pre-PCR processing procedures have been developed to remove or reduce the effects of PCR inhibitors. Pre-PCR processing comprises all steps prior to the detection of PCR products, that is, sampling, sample preparation, and deoxyribonucleic acid (DNA) amplification. The aim of pre-PCR processing is to convert a complex biological sample with its target nucleic acids/cells into PCR-amplifiable samples by combining sample preparation and amplification conditions. Several different pre-PCR processing strategies are used: (1) optimization of the DNA amplification conditions by the use of alternative DNA polymerases and/or amplification facilitators, (2) optimization of the sample preparation method, (3) optimization of the sampling method, and (4) combinations of the different strategies. This review describes different pre-PCR processing strategies to circumvent PCR inhibition to allow accurate and precise DNA amplification.

  3. Mathematical analysis of the real time array PCR (RTA PCR) process

    NARCIS (Netherlands)

    Dijksman, J.F.; Pierik, A.

    2012-01-01

    Real time array PCR (RTA PCR) is a recently developed biochemical technique that measures amplification curves (like with quantitative real time Polymerase Chain Reaction (qRT PCR)) of a multitude of different templates in a sample. It combines two different methods in order to profit from the adva

  4. Transmission of Helicobacter pyori in an animal model.

    Science.gov (United States)

    Cellini, L; Marzio, L; Ferrero, G; Del Vino, A; Di Campli, E; Grossi, L; Toracchio, S; Artese, L

    2001-01-01

    An experimental murine model was studied to evaluate the orogastrointestinal colonization of Helicobacter pylori and the animal-to-animal transmission. Balb/C mice were infected with H. pylori and housed with uninoculated mice in cages with and without a grate on the floor. Mice were killed after 7, 14, 30, and 45 days, and samples from the esophagus, stomach, small intestine, colon, and rectum were analyzed for H. pylori by PCR and immunohistochemistry and for histological changes. Bacterial colonization was assessed also by culture from stomach samples. H. pylori was cultured by stomach samples of infected mice at 7, 14, and 30 days. Using PCR and immunohistochemistry, H. pylori was detected in inoculated and uninoculated mice in all areas examined, with an high percentage of positive samples in the esophagus and stomach. Moreover transmission was detected, without differences, regardless of whether mice were housed with or without a grate on the floor, supporting an orooral animal transmission.

  5. Polymerase chain reaction and real-time PCR for diagnosing of Leishmania infantum chagasi in dogs.

    Science.gov (United States)

    Ramos, Rafael Antonio do Nascimento; Ramos, Carlos Alberto do Nascimento; Jusi, Márcia Mariza Gomes; de Araújo, Flábio Ribeiro; Machado, Rosangela Zacarias; Faustino, Maria Aparecida da Glória; Alves, Leucio Câmara

    2012-01-01

    The importance of dogs as a reservoir for Leishmania infantumchagasi in urban environments has stimulated numerous studies assessing diagnostic techniques. When performed properly, such procedures are an important step in preventing leishmaniasis in humans. Molecular methods have become prominent for this purpose. The aim of the present study was to determine the performance of the polymerase chain reaction (PCR) and real-time PCR (qPCR) for diagnosing of canine visceral leishmaniasis (CVL) using different biological samples. For this, 35 dogs from an area endemic for CVL were used. Bone marrow aspirate and lymph node and spleen fragments from these dogs were used for the molecular diagnosis. In the present study, qPCR was able to detect a greater number of positive animals than seen with PCR. Among the different biological samples used, there was no significant difference in L. infantumchagasi DNA detection between PCR and qPCR. However, considering that lymph nodes are easy to acquire, these can be considered to be the best samples for making molecular diagnoses of L. infantum chagasi infection.

  6. Simultaneous detection and differentiates of Brucella abortus and Brucella melitensis by combinatorial PCR

    Institute of Scientific and Technical Information of China (English)

    Reza Mirnejad; Reza Hosseini Doust; Reza Kachuei; Seied Mojtaba Mortazavi; Mehdi Khoobdel; Ali Ahamadi

    2012-01-01

    Objective:To evaluate simultaneous detection and differentiates of Brucella abortus(B. abortus) and Brucella melitensis (B. melitensis) through the combinatorial PCR method. Methods:This study was designed using three primers that could simultaneously identify and differentiate two major species of pathogenic Brucella in humans and animals. Identification and differentiation of each species using the size of the PCR product were determined. To determine the specificity of the method, bacteria close to the genus Brucella were used. Finally, to confirm PCR products, In addition to the products sequence, RFLP was performed on PCR products using restriction enzymes. Results:The method of optimized combinatorial PCR in this study could simultaneously detect and differentiate B. abortus and B. melitensis with high specificity and sensitivity in clinical samples. Differentiation of species is based on the resulting bands;therefore, the band 494 bp for B. abortus and 733 bp for B. melitensis were obtained. RFLP and sequencing results confirmed PCR results. Conclusions:The results of this study shows that without routine diagnostic methods such as culture and serology tests, using the molecular method of combinatorial PCR, important species of Brucella can be simultaneously identified and differentiated in clinical samples.

  7. Comparison of Microscopy and PCR-RFLP for detection of Anaplasma marginale in carrier cattle

    Directory of Open Access Journals (Sweden)

    P Shayan

    2010-09-01

    Full Text Available Background and Objectives: In Iran, anaplasmosis is normally diagnosed with traditional Giemsa staining method. This is not applicable for identification of the carrier animals. The aim of this study was to compare the detection of Anaplasma marginale in two different numbers of microscopic fields (50 and 100 using conventional Giemsa staining method compared with the PCR-RFLP technique."nMaterials and Methods: In this study, examinations were performed on 150 blood samples from cattle without clinical signs. Sensitivity and specificity of two microscopic fields (50 and 100 fields were compared with A. marginale specific PCR-RFLP. The degree of agreement between PCR-RFLP and the two microscopic tests was determined by Kappa (κ values with 95% confidence intervals."nResults: PCR-RFLP showed that 58 samples were A. marginale, while routine microscopy showed erythrocytes harboring Anaplasma like structures in 16 and 75 blood samples determined in 50 and 100 microscopic fields respectively. Examination of 50 and 100 microscopic fields showed 25.8% and 91.4% sensitivity and 99% and 76.1% specificity compared to 100% sensitivity and specificity by PCR-RFLP. The Kappa coefficient between PCR-RFLP and Microscopy (50 fields indicated a fair level of agreement (0.29. The Kappa coefficient between PCR-RFLP and Microscopy (100 fields indicated a good level of agreement (0.64"nConclusion: Our results showed that the microscopic examination remains the convenient technique for day-to-day diagnosis of clinical cases in the laboratory but for the detection of carrier animal with low bacteremia, microscopy with 100 fields is preferable to Microscopy with 50 fields and molecular methods such as PCR-RFLP can be used as a safe method for identifying cattle persistently infected with A. marginale.

  8. Animal transportation networks.

    Science.gov (United States)

    Perna, Andrea; Latty, Tanya

    2014-11-01

    Many group-living animals construct transportation networks of trails, galleries and burrows by modifying the environment to facilitate faster, safer or more efficient movement. Animal transportation networks can have direct influences on the fitness of individuals, whereas the shape and structure of transportation networks can influence community dynamics by facilitating contacts between different individuals and species. In this review, we discuss three key areas in the study of animal transportation networks: the topological properties of networks, network morphogenesis and growth, and the behaviour of network users. We present a brief primer on elements of network theory, and then discuss the different ways in which animal groups deal with the fundamental trade-off between the competing network properties of travel efficiency, robustness and infrastructure cost. We consider how the behaviour of network users can impact network efficiency, and call for studies that integrate both network topology and user behaviour. We finish with a prospectus for future research.

  9. A northern animal kingdom

    Institute of Scientific and Technical Information of China (English)

    RainerThomm

    2005-01-01

    I began photographing wild animals at Baiquan in 2002,what is really propelling me to go back time and time again,though,is the unforgettable experience of tracking down and getting shots of red foxes and shika.

  10. [Alternatives to animal testing].

    Science.gov (United States)

    Fabre, Isabelle

    2009-11-01

    The use of alternative methods to animal testing are an integral part of the 3Rs concept (refine, reduce, replace) defined by Russel & Burch in 1959. These approaches include in silico methods (databases and computer models), in vitro physicochemical analysis, biological methods using bacteria or isolated cells, reconstructed enzyme systems, and reconstructed tissues. Emerging "omic" methods used in integrated approaches further help to reduce animal use, while stem cells offer promising approaches to toxicologic and pathophysiologic studies, along with organotypic cultures and bio-artificial organs. Only a few alternative methods can so far be used in stand-alone tests as substitutes for animal testing. The best way to use these methods is to integrate them in tiered testing strategies (ITS), in which animals are only used as a last resort.

  11. Animal culture: chimpanzee conformity?

    Science.gov (United States)

    van Schaik, Carel P

    2012-05-22

    Culture-like phenomena in wild animals have received much attention, but how good is the evidence and how similar are they to human culture? New data on chimpanzees suggest their culture may even have an element of conformity.

  12. Animal models of scoliosis.

    Science.gov (United States)

    Bobyn, Justin D; Little, David G; Gray, Randolph; Schindeler, Aaron

    2015-04-01

    Multiple techniques designed to induce scoliotic deformity have been applied across many animal species. We have undertaken a review of the literature regarding experimental models of scoliosis in animals to discuss their utility in comprehending disease aetiology and treatment. Models of scoliosis in animals can be broadly divided into quadrupedal and bipedal experiments. Quadrupedal models, in the absence of axial gravitation force, depend upon development of a mechanical asymmetry along the spine to initiate a scoliotic deformity. Bipedal models more accurately mimic human posture and consequently are subject to similar forces due to gravity, which have been long appreciated to be a contributing factor to the development of scoliosis. Many effective models of scoliosis in smaller animals have not been successfully translated to primates and humans. Though these models may not clarify the aetiology of human scoliosis, by providing a reliable and reproducible deformity in the spine they are a useful means with which to test interventions designed to correct and prevent deformity.

  13. [Spuriously unhealthy animal fats].

    Science.gov (United States)

    Cichosz, Grazyna; Czeczot, Hanna

    2011-11-01

    Animal fats are generally considered as a source of saturated fatty acids and cholesterol, identified with arteriosclerosis and its clinical complications (cardiovascular diseases with heart attack, stroke, cerebral claudication). The real reason of arteriosclerosis are inflammation states of blood vessel endothelium caused by oxidative stress, hiperhomocysteinemia, hipertrigliceridemia, presence of artificial trans isomers and excess of eicosanoids originated from poliunsaturated fatty acids n-6. Present status of science proves that both saturated fatty acids and cholesterol present in animal food can not cause inflammation state. Moreover, animal fats are source of antioxidants active both in food and in human organism. Due to high oxidative stability animal fats do not make threat to human health. Milk fat, though high content of saturated fatty acids and cholesterol, possesses comprehensive pro-health activity--against arteriosclerosis and cancerogenesis.

  14. Animal-free toxicology

    DEFF Research Database (Denmark)

    Knudsen, Lisbeth E

    2013-01-01

    Human data on exposure and adverse effects are the most appropriate for human risk assessment, and modern toxicology focuses on human pathway analysis and the development of human biomarkers. Human biomonitoring and human placental transport studies provide necessary information for human risk...... assessment, in accordance with the legislation on chemical, medicine and food safety. Toxicology studies based on human mechanistic and exposure information can replace animal studies. These animal-free approaches can be further supplemented by new in silico methods and chemical structure......-activity relationships. The inclusion of replacement expertise in the international Three Rs centres, the ongoing exploration of alternatives to animal research, and the improvement of conditions for research animals, all imply the beginning of a paradigm shift in toxicology research toward the use of human data....

  15. Computer animation of clouds

    Energy Technology Data Exchange (ETDEWEB)

    Max, N.

    1994-01-28

    Computer animation of outdoor scenes is enhanced by realistic clouds. I will discuss several different modeling and rendering schemes for clouds, and show how they evolved in my animation work. These include transparency-textured clouds on a 2-D plane, smooth shaded or textured 3-D clouds surfaces, and 3-D volume rendering. For the volume rendering, I will present various illumination schemes, including the density emitter, single scattering, and multiple scattering models.

  16. Trade, Environment & Animal Welfare

    DEFF Research Database (Denmark)

    Morrison, Peter; Nielsen, Laura

    2013-01-01

    Regulation of animal welfare and the environment under the WTO GATT and GATS Agreements - including introduction of the innovative idea of limiting consumption abroad (mode 2) for e.g. bull fights.......Regulation of animal welfare and the environment under the WTO GATT and GATS Agreements - including introduction of the innovative idea of limiting consumption abroad (mode 2) for e.g. bull fights....

  17. On Animal Metaphor

    Institute of Scientific and Technical Information of China (English)

    李凡凡

    2007-01-01

    Nowadays it is common to talk about metaphor. In fact, metaphor is a kind of comparison. Because of comparison and association,familiar objects become strange and glamorous. Animal metaphors can involve either nominal form or verb forms. A person's crying may be called barking. A woman may be called a cat, or a goose, etc. Animal metaphor is connected tightly with our life and helps language development. We can utilize them to make our life and languages more colorful.

  18. Snow White Trench (Animation)

    Science.gov (United States)

    2008-01-01

    [figure removed for brevity, see original site] Click on image for animation This animation shows the evolution of the trench called 'Snow White' that NASA's Phoenix Mars Lander began digging on the 22nd Martian day of the mission after the May 25, 2008, landing. The Phoenix Mission is led by the University of Arizona, Tucson, on behalf of NASA. Project management of the mission is by NASA's Jet Propulsion Laboratory, Pasadena, Calif. Spacecraft development is by Lockheed Martin Space Systems, Denver.

  19. Cytogenetics in animal production

    OpenAIRE

    2010-01-01

    Cytogenetics applied to domestic animals is a useful biotechnology to be applied in the genetic improvement of livestock. Indeed, it can be used to select reproducers free chromosome abnormalities which are responsible for abnormal body conformation (aneuploidy), lower fertility (balanced chromosome abnormalities) or sterility (sex chromosome abnormalities). Cytogenetics may also be applied to assess environmental pollution by studying animals living in hazardous areas and using them as biolo...

  20. Real-Time PCR (RT-PCR) Assays for Burkholderia mallei and B. pseudomallei

    Science.gov (United States)

    2005-10-01

    1 Real - time PCR (RT-PCR) Assays for Burkholderia mallei and B. pseudomallei Vipin K. Rastogi1, Tu-chen Cheng1, Lisa Collins1 and Jennifer Bagley2 1...A 3. DATES COVERED - 4. TITLE AND SUBTITLE Real - time PCR (RT-PCR) Assays for Burkholderia mallei and B.pseudomallei 5a. CONTRACT NUMBER 5b...risk. There is currently no real - time PCR assay for detection of both of these pathogens. Primers and probes corresponding to specific genomic regions

  1. Material Biocompatibility for PCR Microfluidic Chips

    KAUST Repository

    Kodzius, Rimantas

    2010-04-23

    As part of the current miniaturization trend, biological reactions and processes are being adapted to microfluidics devices. PCR is the primary method employed in DNA amplification, its miniaturization is central to efforts to develop portable devices for diagnostics and testing purposes. A problem is the PCR-inhibitory effect due to interaction between PCR reagents and the surrounding environment, which effect is increased in high-surface-are-to-volume ration microfluidics. In this study, we evaluated the biocompatibility of various common materials employed in the fabrication of microfluidic chips, including silicon, several kinds of silicon oxide, glasses, plastics, wax, and adhesives. Two-temperature PCR was performed with these materials to determine their PCR-inhibitory effect. In most of the cases, addition of bovine serum albumin effectively improved the reaction yield. We also studied the individual PCR components from the standpoint of adsorption. Most of the materials did not inhibit the DNA, whereas they did show noticeable interaction with the DNA polymerase. Our test, instead of using microfluidic devices, can be easily conducted in common PCR tubes using a standard bench thermocycler. Our data supports an overview of the means by which the materials most bio-friendly to microfluidics can be selected.

  2. Design and Assessment of a Real Time Reverse Transcription-PCR Method to Genotype Single-Stranded RNA Male-Specific Coliphages (Family Leviviridae).

    Science.gov (United States)

    A real-time, reverse transcription-PCR (RT-qPCR) assay was developed to differentiate the four genogroups of male-specific ssRNA coliphages (FRNA) (family Leviviridae). As FRNA display a trend of source-specificity (human sewage or animal waste) at the genogroup level, this assa...

  3. Quantitative DNA Analysis Using Droplet Digital PCR.

    Science.gov (United States)

    Vossen, Rolf H A M; White, Stefan J

    2017-01-01

    Droplet digital PCR (ddPCR) is based on the isolated amplification of thousands of individual DNA molecules simultaneously, with each molecule compartmentalized in a droplet. The presence of amplified product in each droplet is indicated by a fluorescent signal, and the proportion of positive droplets allows the precise quantification of a given sequence. In this chapter we briefly outline the basis of ddPCR, and describe two different applications using the Bio-Rad QX200 system: genotyping copy number variation and quantification of Illumina sequencing libraries.

  4. Evaluation of PCR methods for detection of Brucella strains from culture and tissues.

    Science.gov (United States)

    Çiftci, Alper; İça, Tuba; Savaşan, Serap; Sareyyüpoğlu, Barış; Akan, Mehmet; Diker, Kadir Serdar

    2017-03-03

    The genus Brucella causes significant economic losses due to infertility, abortion, stillbirth or weak calves, and neonatal mortality in livestock. Brucellosis is still a zoonosis of public health importance worldwide. The study was aimed to optimize and evaluate PCR assays used for the diagnosis of Brucella infections. For this aim, several primers and PCR protocols were performed and compared with Brucella cultures and biological material inoculated with Brucella. In PCR assays, genus- or species-specific oligonucleotide primers derived from 16S rRNA sequences (F4/R2, Ba148/928, IS711, BruP6-P7) and OMPs (JPF/JPR, 31ter/sd) of Brucella were used. All primers except for BruP6-P7 detected the DNA from reference Brucella strains and field isolates. In spiked blood, milk, and semen samples, F4-R2 primer-oriented PCR assays detected minimal numbers of Brucella. In spiked serum and fetal stomach content, Ba148/928 primer-oriented PCR assays detected minimal numbers of Brucella. Field samples collected from sheep and cattle were examined by bacteriological methods and optimized PCR assays. Overall, sensitivity of PCR assays was found superior to conventional bacteriological isolation. Brucella DNA was detected in 35.1, 1.1, 24.8, 5.0, and 8.0% of aborted fetus, blood, milk, semen, and serum samples by PCR assays, respectively. In conclusion, PCR assay in optimized conditions was found to be valuable in sensitive and specific detection of Brucella infections of animals.

  5. Tamizaje de deleciones en pacientes con distrofia muscular de Duchenne (DMD) o Becker-Kiener (BMD) mediante PCR multiplex en Costa Rica, 1998-2000

    OpenAIRE

    Sancho-Fernández, Vanessa M.; Saborio, Manuel; de Céspedes-Montealegre, Carlos; Azofeifa-Navas, Jorge

    2001-01-01

    Artículo científico -- Universidad de Costa Rica, Instituto de Investigaciones en Salud. 2001 Objetivo: Iniciar los estudios genético moleculares sobre las distrofinopatías en Costa Rica. Materiales y Métodos: Treinta y un pacientes varones, diagnosticados con distrofia muscular, que podrían ser distrofinopáticos fueron reevaluados clínicamente. Veintitrés mostraron un fenotipo de DMD y dos de BMD. Seis no mostraron síntomas definitivos de distrofinopatías. ADN de los pacientes fue analiza...

  6. Comparison of LAMP and PCR for molecular mass screening of sand flies for Leishmania martiniquensis infection

    Science.gov (United States)

    Tiwananthagorn, Saruda; Kato, Hirotomo; Yeewa, Ranchana; Muengpan, Amontip; Polseela, Raxsina; Leelayoova, Saovanee

    2017-01-01

    BACKGROUND Leishmaniasis caused by Leishmania martiniquensis infection has been reported in human and domestic animals of Martinique Island, Germany, Switzerland, USA, Myanmar and Thailand. The peculiar clinical features of disseminated cutaneous and visceral forms co-existence render the urgent need of specific diagnostic tool to identify the natural sand fly vectors for effective prevention and control strategies. Loop-mediated isothermal amplification (LAMP) of 18S rRNA gene as well as polymerase chain reaction (PCR) of minicircle kinetoplast DNA gene (PCR-mkDNA) have never been applied to detect L. martiniquensis and L. siamensis in sand fly vectors. OBJECTIVE The present study was aimed to validate malachite green-LAMP (MG-LAMP) and PCR-mkDNA techniques to detect L. martiniquensis in sand fly vectors, compared with the conventional PCR of internal transcribed spacer 1 (PCR-ITS1). METHODS We compared the validity of LAMP of 18S rRNA gene and PCR-mkDNA, to PCR-ITS1 in simulation model of L. martiniquensis infection in Sergentomyia gemmea sand flies. Attributable to the sensitivity and specificity, PCR-mkDNA was consecutively applied to detect L. martiniquensis in 380 female sand fly individuals captured in the newly identified affected region of Lamphun Province, Thailand. FINDINGS AND MAIN CONCLUSIONS Results showed that PCR-mkDNA could detect at least one promastigote per sand fly, which was 10-time superior to LAMP and PCR-ITS1. In addition, PCR-mkDNA was more specific, able to differentiate L. martiniquensis from other viscerotropic Leishmania species, such as L. siamensis, L. (L.) donovani, and L. (L.) infantum. Consecutively, mass screening of L. martiniquensis in 380 female sand fly individuals by PCR-mkDNA was implemented in a new affected area of Thailand where a patient with leishmaniasis/HIV co-infection resides; however Leishmania DNA was undetected. In conclusion, PCR-mkDNA is a promising tool for molecular mass screening of L. martiniquensis

  7. Becoming Sheep, Becoming Animal

    DEFF Research Database (Denmark)

    Grum, Charlotte; Svabo, Connie

    2016-01-01

    Proposal for Performance Research, in response to the call Turning Animal: As a part of a 2015 group exhibition exploring the history and local myths of a woman living in a Danish heath landscape 150 years ago, artist Charlotte Grum connected herself to a live sheep for 4 hours a day, 5 days a we...... support the written account – together with graphic figurations of the many human and non-human actors playing a part of the mattering of “Becoming Sheep”, with an equal intention of performing multiple positions within and through the text......., for 5 weeks, turning the two into a hybrid relational assemblage, intra-acting and becoming with the heath habitat, the other by-passing human and non-human animals, the changing weather and their fluctuating biological needs. She wanted to explore the discursive and material effects of a site......-specific human-nonhuman animal intra-action, to challenge the gendered and anthropocentric reading of a particular historical subject and to explore the messy constituents of the very categories of women and animals. In general she is occupied with how to animate and perform the intra-active entanglement...

  8. Animal models for osteoporosis

    Science.gov (United States)

    Turner, R. T.; Maran, A.; Lotinun, S.; Hefferan, T.; Evans, G. L.; Zhang, M.; Sibonga, J. D.

    2001-01-01

    Animal models will continue to be important tools in the quest to understand the contribution of specific genes to establishment of peak bone mass and optimal bone architecture, as well as the genetic basis for a predisposition toward accelerated bone loss in the presence of co-morbidity factors such as estrogen deficiency. Existing animal models will continue to be useful for modeling changes in bone metabolism and architecture induced by well-defined local and systemic factors. However, there is a critical unfulfilled need to develop and validate better animal models to allow fruitful investigation of the interaction of the multitude of factors which precipitate senile osteoporosis. Well characterized and validated animal models that can be recommended for investigation of the etiology, prevention and treatment of several forms of osteoporosis have been listed in Table 1. Also listed are models which are provisionally recommended. These latter models have potential but are inadequately characterized, deviate significantly from the human response, require careful choice of strain or age, or are not practical for most investigators to adopt. It cannot be stressed strongly enough that the enormous potential of laboratory animals as models for osteoporosis can only be realized if great care is taken in the choice of an appropriate species, age, experimental design, and measurements. Poor choices will results in misinterpretation of results which ultimately can bring harm to patients who suffer from osteoporosis by delaying advancement of knowledge.

  9. The potential advantages of digital PCR for clinical virology diagnostics.

    Science.gov (United States)

    Hall Sedlak, Ruth; Jerome, Keith R

    2014-05-01

    Digital PCR (dPCR), a new nucleic acid amplification technology, offers several potential advantages over real-time or quantitative PCR (qPCR), the current workhorse of clinical molecular virology diagnostics. Several studies have demonstrated dPCR assays for human cytomegalovirus or HIV, which give more precise and reproducible results than qPCR assays without sacrificing sensitivity. Here we review the literature comparing dPCR and qPCR performance in viral molecular diagnostic assays and offer perspective on the future of dPCR in clinical virology diagnostics.

  10. Calibrated user-friendly reverse transcriptase-PCR assay

    DEFF Research Database (Denmark)

    Bor, M V; Sørensen, B S; Rammer, P;

    1998-01-01

    We report a competitive reverse transcriptase-PCR (RT-PCR) assay and a calibrated user-friendly RT-PCR assay (CURT-PCR) for epidermal growth factor receptor (EGFR) mRNA. A calibrator was prepared from isolated rat liver RNA, and the amount of EGFR mRNA was determined by competitive RT-PCR. In CUR...

  11. Real time PCR. Application in dengue studies

    Directory of Open Access Journals (Sweden)

    Jeanette Prada-Arismendy

    2011-06-01

    Full Text Available PCR (polymerase chain reaction is a routinely used tool in every diagnostic and research laboratory. This technique has been used in detection of mutations and pathogens, forensic investigation, and even is the base tool for human genome sequencing. A modification of PCR technique, real time PCR, allows the quantification of nucleic acids with higher sensibility, specificity and reproducibility. This article is intended to clarify the foundations of real-time PCR, using an application model for virology. In the actual work, it was quantified the viral load of dengue virus serotype 2 produced from infected murine macrophages; the obtained results in this work established that murine strain BALB/c presents a greater susceptibility to dengue virus infection, which establishes BALB/c murine strain as a best model of study for investigation of dengue virus infection physiopathology.

  12. Universally Primed PCR (UP-PCR) and its applications for taxonomy in Trichoderma

    Institute of Scientific and Technical Information of China (English)

    Mette Lübeck

    2004-01-01

    @@ Universally Primed PCR (UP-PCR) is a PCR fingerprinting method that has demonstrated its applicability in different aspects of mycology. These applications constitute analysis of genome structures, identification of species, analysis of population and species diversity, revealing of genetic relatedness at infra-and inter-species level, and identification of UP-PCR markers at different taxonomic levels (strain, group and/or species) . A further development of the UP-PCR technique is an UP-PCR product cross hybridisation assay that facilitates investigation of sequence similarity (homology) of UP-PCR products and grouping of strains into UP-PCR hybridisation groups. This separates the strains into entities with high genetic similarity (DNA homology) . UP-PCR has been used as an aid in taxonomy and species delineation, and to monitor biocontrol strains following their release into the environment by fingerprint characterisation of pure cultures and through direct detection in soil by amplification of UP-PCR-derived SCAR markers. The technique has been applied to Trichoderma strains in particularly with the aims of strain recognition and classification.

  13. Phoenix Lidar Operation Animation

    Science.gov (United States)

    2008-01-01

    [figure removed for brevity, see original site] Click on image for animation This is an animation of the Canadian-built meteorological station's lidar, which was successfully activated on Sol 2. The animation shows how the lidar is activated by first opening its dust cover, then emitting rapid pulses of light (resembling a brilliant green laser) into the Martian atmosphere. Some of the light then bounces off particles in the atmosphere, and is reflected back down to the lidar's telescope. This allows the lidar to detect dust, clouds and fog. The Phoenix Mission is led by the University of Arizona, Tucson, on behalf of NASA. Project management of the mission is by NASA's Jet Propulsion Laboratory, Pasadena, Calif. Spacecraft development is by Lockheed Martin Space Systems, Denver.

  14. Animals and ICE

    DEFF Research Database (Denmark)

    van Hemmen, J Leo; Christensen-Dalsgaard, Jakob; Carr, Catherine E

    2016-01-01

    experimental and mathematical foundation, it is known that there is a low-frequency regime where the internal time difference (iTD) as perceived by the animal may well be 2-5 times higher than the external ITD, the interaural time difference, and that there is a frequency plateau over which the fraction i......TD/ITD is constant. There is also a high-frequency regime where the internal level (amplitude) difference iLD as perceived by the animal is much higher than the interaural level difference ILD measured externally between the two ears. The fundamental tympanic frequency segregates the two regimes. The present special...... issue devoted to "internally coupled ears" provides an overview of many aspects of ICE, be they acoustic, anatomical, auditory, mathematical, or neurobiological. A focus is on the hotly debated topic of what aspects of ICE animals actually exploit neuronally to localize a sound source....

  15. Theriocide: Naming Animal Killing

    Directory of Open Access Journals (Sweden)

    Piers Beirne

    2014-08-01

    Full Text Available In this essay I recommend ‘theriocide’ as the name for those diverse human actions that cause the deaths of animals. Like the killing of one human by another, theriocide may be socially acceptable or unacceptable, legal or illegal. It may be intentional or unintentional and may involve active maltreatment or passive neglect. Theriocide may occur one-on-one, in small groups or in large-scale social institutions. The numerous and sometimes intersecting sites of theriocide include intensive rearing regimes; hunting and fishing; trafficking; vivisection; militarism; pollution; and human-induced climate change. If the killing of animals by humans is as harmful to them as homicide is to humans, then the proper naming of such deaths offers a remedy, however small, to the extensive privileging of human lives over those of other animals. Inevitably, the essay leads to a shocking question: Is theriocide murder?

  16. Animal Poetry and Empathy

    Directory of Open Access Journals (Sweden)

    Tirza Brüggemann

    2017-04-01

    Full Text Available This article discusses how our ideas of empathy are influenced by the dichotomy of mind versus body, also known as Cartesian dualism. Within the aesthetic field, this dichotomy is seen when researchers define narrative empathy as imaginatively reconstructing the fictional character’s thoughts and feelings. Conversely, the empathy aroused by a non-narrative work of art is seen as an unconscious bodily mirroring of movements, postures or moods. Thinking dualistically does not only have consequences for what we consider human nature; it also affects our view on animals. To show the untenability of dualistic thinking, this article focuses on the animal poetry genre. Using the ideas of the French phenomenologist Maurice Merleau-Ponty, I analyze two animal poems: “Inventing a Horse” by Meghan O’Rourke and “Spermaceti” by Les Murray. The analysis of these two poems suggests that the presiding ideas about aesthetic empathy and empathy in general need re-evaluation.

  17. Programación por metas Energía alternativa mediante biomasa.

    Directory of Open Access Journals (Sweden)

    Guerrero Casas, Flor María

    2003-01-01

    Full Text Available En este trabajo se presenta un modelo multicriterio de localización de centrales de generación de energía eléctrica mediante biomasa. Los objetivos considerados son: (1 minimizar el coste total de la operación, (2 maximizar la producción de electricidad obtenida, (3 maximizar la distancia entre plantas, (4 maximizar la aceptación social y (5 establecer las plantas o ampliaciones en aquellos lugares donde exista una mayor predisposición por parte de las administraciones locales. Finalmente, se concluye con una aplicación práctica mediante programación por metas ponderadas para la región andaluza, considerando los residuos procedentes del olivar como fuente de energía.

  18. Transgene detection by digital droplet PCR.

    Directory of Open Access Journals (Sweden)

    Dirk A Moser

    Full Text Available Somatic gene therapy is a promising tool for the treatment of severe diseases. Because of its abuse potential for performance enhancement in sports, the World Anti-Doping Agency (WADA included the term 'gene doping' in the official list of banned substances and methods in 2004. Several nested PCR or qPCR-based strategies have been proposed that aim at detecting long-term presence of transgene in blood, but these strategies are hampered by technical limitations. We developed a digital droplet PCR (ddPCR protocol for Insulin-Like Growth Factor 1 (IGF1 detection and demonstrated its applicability monitoring 6 mice injected into skeletal muscle with AAV9-IGF1 elements and 2 controls over a 33-day period. A duplex ddPCR protocol for simultaneous detection of Insulin-Like Growth Factor 1 (IGF1 and Erythropoietin (EPO transgenic elements was created. A new DNA extraction procedure with target-orientated usage of restriction enzymes including on-column DNA-digestion was established. In vivo data revealed that IGF1 transgenic elements could be reliably detected for a 33-day period in DNA extracted from whole blood. In vitro data indicated feasibility of IGF1 and EPO detection by duplex ddPCR with high reliability and sensitivity. On-column DNA-digestion allowed for significantly improved target detection in downstream PCR-based approaches. As ddPCR provides absolute quantification, it ensures excellent day-to-day reproducibility. Therefore, we expect this technique to be used in diagnosing and monitoring of viral and bacterial infection, in detecting mutated DNA sequences as well as profiling for the presence of foreign genetic material in elite athletes in the future.

  19. Animal violence demystified

    Directory of Open Access Journals (Sweden)

    Deepa Natarajan

    2010-04-01

    Full Text Available Violence has been observed in humans and animals alike, indicating its evolutionary/ biological significance. However, violence in animals has often been confounded with functional forms of aggressive behavior. Currently, violence in animals is identified primarily as either a quantitative behavior (an escalated, pathological and abnormal form of aggression characterized primarily by short attack latencies, and prolonged and frequent harm-oriented conflict behaviors or a qualitative one (characterized by attack bites aimed at vulnerable parts of the opponent’s body and context independent attacks regardless of the environment or the sex and type of the opponent. Identification of an operational definition for violence thus not only helps in understanding its potential differences from adaptive forms of aggression but also in the selection of appropriate animal models for both. To begin with, we address this issue theoretically by drawing parallels from research on aggression and appeasement in humans and other animals. We also provide empirical evidences for violence in mice selected for high aggression by comparing our findings with other currently available potentially violent rodent models. The following violence-specific features namely 1. Display of low levels of pre-escalatory/ritualistic behaviors. 2. Immediate and escalated offense durations with low withdrawal rates despite the opponent’s submissive supine and crouching/defeat postures. 3. Context independent indiscriminate attacks aimed at familiar/unfamiliar females, anaesthetized males and opponents and in neutral environments. 4. Orientation of attack-bites toward vulnerable body parts of the opponent resulting in severe wounding 5. Low pre-frontal serotonin (5-HT levels upon repeated aggression. 6. Low basal heart rates and hyporesponsive hypothalamus-pituitary-adrenocortical (HPA axis were identified uniquely in the short attack latency (SAL mice suggesting a qualitative

  20. Brucella abortus is Prevalent in Both Humans and Animals in Bangladesh.

    Science.gov (United States)

    Rahman, A K M A; Saegerman, C; Berkvens, D; Melzer, F; Neubauer, H; Fretin, D; Abatih, E; Dhand, N; Ward, M P

    2017-01-09

    To determine the role of different Brucella (B.) spp. in Bangladesh, 62 animal samples and 500 human sera were tested. Animal samples from cattle, goats and sheep (including milk, bull semen, vaginal swabs and placentas) were cultured for Brucella spp. Three test-positive human sera and all animal samples were screened by Brucella genus-specific real-time PCR (RT-PCR), and positive samples were then tested by IS711 RT-PCR to detect B. abortus and B. melitensis DNA. Only B. abortus DNA was amplified from 13 human and six animal samples. This is the first report describing B. abortus as the aetiological agent of brucellosis in occupationally exposed humans in Bangladesh. Of note is failure to detect B. melitensis DNA, the species most often associated with human brucellosis worldwide. Further studies are required to explore the occurrence of Brucella melitensis in Bangladesh.

  1. Development of a real-time SYBR Green PCR assay for the rapid detection of Dermatophilus congolensis

    Science.gov (United States)

    Martínez, Remigio; Benitez-Medina, José Manuel; Risco, David; García, Waldo Luis; Rey, Joaquín; Alonso, Juan Manuel; de Mendoza, Javier Hermoso

    2013-01-01

    Methods such as real time (RT)-PCR have not been developed for the rapid detection and diagnosis of Dermatophilus (D.) congolensis infection. In the present study, a D. congolensis-specific SYBR Green RT-PCR assay was evaluated. The detection limit of the RT-PCR assay was 1 pg of DNA per PCR reaction. No cross-reaction with nucleic acids extracted from Pseudomonas aeruginosa, Mycobacterium tuberculosis, Staphylococcus aureus, or Austwickia chelonae was observed. Finally, the RT-PCR assay was used to evaluate clinical samples collected from naturally infected animals with D. congolensis. The results showed that this assay is a fast and reliable method for diagnosing dermatophilosis. PMID:23820221

  2. Use of a Mycoplasma suis-PCR protocol for screening a population of captive peccaries (Tayassu tajacu and Tayassu pecari).

    Science.gov (United States)

    Vieira, Rafael Felipe da Costa; Molento, Marcelo Beltrão; Guimarães, Ana Marcia Sa; Santos, Andrea Pires Dos; Bonat, Marcelo; Javorouski, Manoel Lucas; Popp, Luciene; Santos, Leonilda Correia Dos; Moraes, Wanderlei; Cubas, Zalmir Silvino; Vieira, Thállitha Samih Wischral Jayme; Vidotto, Odilon; Filho, Ivan Roque Barros; Biondo, Alexander Welker; Messick, Joanne Belle

    2011-01-01

    Mycoplasma suis is a hemotropic bacteria of red blood cells and the causative agent of swine eperythrozoonosis. Diagnosis of infection may be reached by direct examination of blood smears; however, the use of polymerase chain reaction (PCR) of the 16S RNA gene of M. suis improves the sensitivity and specificity of detection. The aim of this study was to screen peccaries (Tayassu tajacu and T. pecari) for M. suis infection using a specific conventional PCR. A total of 28 blood samples from captive collared and white-lipped peccaries were collected, DNA extracted and a specific M. suis PCR assay performed. All samples were negatives by both blood smear examination and PCR testing. To verify the presence of amplifiable DNA, PCR for beta-actin gene was performed in all samples. This study was part of an active surveillance program, which is crucial for monitoring animal health status, particularly in wildlife species.

  3. Animation of MARDI Instrument

    Science.gov (United States)

    2008-01-01

    [figure removed for brevity, see original site] Click on image to view the animation This animation shows a zoom into the Mars Descent Imager (MARDI) instrument onboard NASA's Phoenix Mars Lander. The Phoenix team will soon attempt to use a microphone on the MARDI instrument to capture sounds of Mars. The Phoenix Mission is led by the University of Arizona, Tucson, on behalf of NASA. Project management of the mission is by NASA's Jet Propulsion Laboratory, Pasadena, Calif. Spacecraft development is by Lockheed Martin Space Systems, Denver.

  4. A naked-eye colorimetric "PCR developer"

    Science.gov (United States)

    Valentini, Paola; Pompa, Pier Paolo

    2016-04-01

    Despite several advances in molecular biology and diagnostics, Polymerase Chain Reaction (PCR) is currently the gold standard for nucleic acids amplification and detection, due to its versatility, low-cost and universality, with estimated market of several billion dollars/year. Nevertheless, PCR still relies on the laborious, time-consuming, and multi-step gel electrophoresis-based detection, which includes gel casting, electrophoretic run, gel staining, and gel visualization. In this work, we propose a "PCR developer", namely a universal one-step, one-tube method, based on controlled aggregation of gold nanoparticles (AuNPs), to detect PCR products by naked eye in few minutes, with no need for any instrumentation. We demonstrated the specificity and sensitivity of the PCR developer on different model targets, suitable for a qualitative detection in real-world diagnostics (i.e., gene rearrangements, genetically modified organisms, and pathogens). The PCR developer proved to be highly specific and ultra-sensitive, discriminating down to few copies of HIV viral DNA, diluted in an excess of interfering human genomic DNA, which is a clinically relevant viral load. Hence, it could be a valuable tool for both academic research and clinical applications.

  5. Miglioramento del profilo biofarmaceutico dell'albendazolo mediante co-cristallizzazione

    OpenAIRE

    Pranzo, Marco Beniamino

    2010-01-01

    Lo scopo del presente lavoro di tesi è stato quello di studiare il possibile miglioramento del profilo biofarmaceutico dell’ albendazolo attraverso una modificazione delle proprietà chimico-fisiche del farmaco stesso. Tale obiettivo è stato perseguito mediante: - ricristallizzazione del farmaco in solventi differenti al fine di isolare eventuali polimorfi; - studio delle interazioni allo stato solido tra il farmaco e vari CCFs al fine di ottenere un co-cristallo con caratteristiche ...

  6. Materiales conductores mediante funcionalización de polímeros con nanomateriales conductores

    OpenAIRE

    Gutierrez, Manuel; Llobera, Andreu; Fernández Sánchez, César; Jiménez Jorquera, Cecilia; Mendoza Gómez, Ernest

    2010-01-01

    La presente invención se refiere un material conductor que comprende un polímero funcionalizado y a un nanomaterial conductor unidos mediante enlace covalente. La invención también se refiere a un procedimiento de obtención del material conductor sin modificar las propiedades ópticas del polímero y a su uso para la fabricación de sensores o electrodos.

  7. Aislamiento de Mycobacterium avium subsp. Paratuberculosis de fecas en rebaños lecheros infectados mediante el Método de Cornell modificado Isolation of Mycobacterium avium subsp. Paratuberculosis from bovine feces of infected dairy herds by the Cornell’s Method modified

    Directory of Open Access Journals (Sweden)

    J. P SOTO

    2002-01-01

    Full Text Available Con la finalidad de aumentar la tasa de aislamiento de Mycobacterium avium subsp. paratuberculosis (Map, a partir de muestras de fecas bovinas, se evaluó un nuevo procedimiento de descontaminación y cultivo de muestras fecales en 250 animales clínicamente sanos, provenientes de 14 rebaños infectados del sur de Chile. Para la descontaminación de las muestras previo al cultivo se utilizó una solución al 0.9% de cloruro de hexadecilpiridinio (HPC y una solución antibiótica con amfotericina B, vancomicina y ácido nalidíxico. Para el aislamiento del agente se utilizó el medio de Herrold con yema de huevo y micobactina J adicionado de una solución antibiótica similar a la empleada para la descontaminación de las muestras. En el 16% (40 de las muestras analizadas fue posible aislar Map con un 7.6% de contaminación con hongos sólo a partir de la octava semana de incubación. La identidad de las cepas aisladas fue confirmada en el 100% de los casos mediante PCR, utilizando partidores específicos para este agente (P90 y P91. La alta tasa de aislamiento, la especificidad del medio de cultivo y la baja tasa de contaminación de los cultivos, durante el prolongado período de incubación, hacen de este procedimiento una buena alternativa de diagnóstico de Paratuberculosis bovinaIn order to improve the isolation rate of Mycobacterium avium subsp. paratuberculosis (Map from bovine feces a new bacteriological procedure for decontamination and cultivation of fecal samples was evaluated in 250 samples collected from asymptomatic animals in 14 infected dairy herds in southern Chile. Before culture all samples were treated with a decontaminant solution containing 0.9% hexadecylpiridinium chloride and an antibiotic solution containing amphotericin B, vancomycin and nalidixic acid. Herrold Egg Yolk Medium (HEYM and mycobactin J supplemented with the same antibiotics mentioned above was used for the isolation of the agent. Map was isolated from 40

  8. Animal Bites of the Hand

    Science.gov (United States)

    ... animals include cats, rodents, rabbits, ferrets, farm animals, monkeys and reptiles. The major concern of all animal ... the tabs at the top (Video, Articles/WEB, Images, JHS, Products/Vendors), or the filters on the ...

  9. Animal Watching: Outdoors and In.

    Science.gov (United States)

    McLure, John W.

    2001-01-01

    Describes using domesticated, wild, or feral animals to teach students about nature and animal behavior. Connections can be made with psychology, economics, genetics, history, art, and other disciplines. The study of animal behavior provides opportunities for harmless student experimentation. (SAH)

  10. Animal Care in the Classroom.

    Science.gov (United States)

    Llewellyn, Gerald C.

    1979-01-01

    Discusses housing facilities for living animals in the classroom or laboratory. The construction of animal cages from materials obtained locally is described. Space recommendations for laboratory animals and cages are also included. (HM)

  11. Determination of PCR efficiency in chelex-100 purified clinical samples and comparison of real-time quantitative PCR and conventional PCR for detection of Chlamydia pneumoniae

    Directory of Open Access Journals (Sweden)

    Jensen Jørgen

    2002-07-01

    Full Text Available Abstract Background Chlamydia pneumoniae infection has been detected by serological methods, but PCR is gaining more interest. A number of different PCR assays have been developed and some are used in combination with serology for diagnosis. Real-time PCR could be an attractive new PCR method; therefore it must be evaluated and compared to conventional PCR methods. Results We compared the performance of a newly developed real-time PCR with a conventional PCR method for detection of C. pneumoniae. The PCR methods were tested on reference samples containing C. pneumoniae DNA and on 136 nasopharyngeal samples from patients with a chronic cough. We found the same detection limit for the two methods and that clinical performance was equal for the real-time PCR and for the conventional PCR method, although only three samples tested positive. To investigate whether the low prevalence of C. pneumoniae among patients with a chronic cough was caused by suboptimal PCR efficiency in the samples, PCR efficiency was determined based on the real-time PCR. Seventeen of twenty randomly selected clinical samples had a similar PCR efficiency to samples containing pure genomic C. pneumoniae DNA. Conclusions These results indicate that the performance of real-time PCR is comparable to that of conventional PCR, but that needs to be confirmed further. Real-time PCR can be used to investigate the PCR efficiency which gives a rough estimate of how well the real-time PCR assay works in a specific sample type. Suboptimal PCR efficiency of PCR is not a likely explanation for the low positivity rate of C. pneumoniae in patients with a chronic cough.

  12. Sampling and Pooling Methods for Capturing Herd Level Antibiotic Resistance in Swine Feces using qPCR and CFU Approaches

    DEFF Research Database (Denmark)

    Schmidt, Gunilla Veslemøy; Mellerup, Anders; Christiansen, Lasse Engbo;

    2015-01-01

    in the same swine fecal samples. The results showed that the qPCR assays were capable of detecting differences in antibiotic resistance levels in individual animals that the coliform bacteria colony forming units (CFU) could not. Also, the qPCR assays more accurately quantified antibiotic resistance genes...... when comparing individual sampling and pooling methods. qPCR on pooled samples was found to be a good representative for the general resistance level in a pig herd compared to the coliform CFU counts. It had significantly reduced relative standard deviations compared to coliform CFU counts in the same...

  13. Development and Application of a PCR Approach for Detection of Bovis, Sheep, Pig, and Chicken Derived Materials in Feedstuff

    Institute of Scientific and Technical Information of China (English)

    LUO Jia-qin; WANG Jia-qi; BU Deng-pan; LI Dan; WANG Li; WEI Hong-yang; ZHOU Ling-yun

    2008-01-01

    A PCR method for detection of bovis, sheep, pig, and chicken derived materials in feedstuff was established, and the existing method was improved according to the research on general primer and species-specific primers. First, general primer designed according to 16S rRNA gene sequence of bovis, sheep, pig, chicken, fish, and horse mtDNA was used for primary detection of animal derived materials in feedstuff. Species-specific primers designed according to conserved sequence of mtDNA of bovis, sheep, pig, and chicken were used for amplification of a 271, 274, 149, and 266 bp fragment, respectively. Further confirmation of the detection result was then carried out. PCR method for detection of animal derived materials in unknown feedstuff was developed by using general primer, relevant PCR system, and PCR condition. Also a PCR method for detection of each species (bovines, sheep, pig, and chicken) was designed by using our species-specific primers. High sensitivity and specificity of our method were confirmed with a minimum detection level of 0.1%. Method for detection of animal derived materials in this research is not only cheap and easy for operation but also precise and reliable results can be obtained. It could be one of the effective methods for the detection of animal derived materials in feedstuff.

  14. Comparison of clinical samples for visceral Leishmaniasis diagnosis in asymptomatic dogs by PCR hybridization

    Energy Technology Data Exchange (ETDEWEB)

    Ferreira, Sidney A.; Ituassu, Leonardo T.; Melo, Maria N. [Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, MG (Brazil). Dept. de Parasitologia], e-mail: saninoalmeida@gmail.com, e-mail: Itituassu@yahoo.com.br, e-mail: melo@icb.ufmg.br; Leite, Rodrigo S.; Andrade, Antero S.R. [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN-CNEN/MG), Belo Horizonte, MG (Brazil)], e-mail: rleite2005@gmail.com, e-mail: antero@cdtn.br

    2009-07-01

    The canine visceral leishmaniasis (CVL) diagnosis still represents a challenge because of complexity of this disease. The aim of present study was to compare different clinical samples for diagnosis of CVL by Polymerase Chain Reaction (PCR) combined with hybridization of {sup 32}P labeled probes. Bone marrow (BM), skin biopsy (SB), peripheral blood (PB) and conjunctival swab (CS) were used in this work. With this purpose 40 asymptomatic dogs, all positive by parasitological test, were obtained. From each animal were collected SB with sterile punches from ear internal surface, 1.0 mL of PB, BM aspirates from sternum and CS from both lower eyelid. Each clinical sample was submitted to suitable DNA purification process and PCR-hybridization assays. The positive results obtained with PCR were 55%, 25%, 30% and 22.5% for CS, BM, SB and PB respectively while the PCR followed by hybridization showed a positivity of 87.5%, 50%, 45% and 27.5% respectively. The hybridization assay was able to increase the PCR positivity in all kinds of clinical samples. The best performance was obtained using CS samples. We concluded that the PCR associated with DNA radioactive probes was a very sensitive tool for diagnosis of CVL in asymptomatic dogs and the CS has an important potential for regular screening of dogs. (author)

  15. Propidium Monoazide Coupled with PCR Predicts Infectivity of Enteric Viruses in Swine Manure and Biofertilized Soil.

    Science.gov (United States)

    Fongaro, Gislaine; Hernández, Marta; García-González, María Cruz; Barardi, Célia Regina Monte; Rodríguez-Lázaro, David

    2016-03-01

    The use of propidium monoazide (PMA) coupled with real-time PCR (RT-qPCR or qPCR for RNA or DNA viruses, respectively) was assessed to discriminate infectious enteric viruses in swine raw manure, swine effluent from anaerobic biodigester (AB) and biofertilized soils. Those samples were spiked either with infectious and heat-inactivated human adenovirus-2 (HAdV-2) or mengovirus (vMC0), and PMA-qPCR/RT-qPCR allowed discriminating inactivated viruses from the infective particles, with significant reductions (>99.9%). Then, the procedure was further assayed to evaluate the presence and stability of two non-cultivable viruses (porcine adenovirus and rotavirus A) in natural samples (swine raw manure, swine effluent from AB and biofertilized soils); it demonstrated viral inactivation during the storage period at 23 °C. As a result, the combination of PMA coupled to real-time PCR can be a promising alternative for prediction of viral infectivity in comparison to more labour-intensive and costly techniques such as animal or tissue-culture infectivity methods, and for those viruses that do not have currently available cell culture techniques.

  16. Quantification of GPCR mRNA using real-time RT-PCR.

    Science.gov (United States)

    Brattelid, Trond; Levy, Finn Olav

    2011-01-01

    Characterisation of G-protein-coupled receptor (GPCR) mRNA expression under normal, different pharmacological and pathological conditions in experimental animal models and human tissue biopsies by quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) is a valuable approach to understand the regulation of GPCR expression. RT-qPCR is specific and sensitive with a broad dynamic range, which allows precise quantification of mRNA species of interest. In addition to measuring the relative levels of mRNA in a tissue or changes in expression levels between groups of genes of interest, RT-qPCR is also used to identify splice variants and single nucleotide polymorphisms (SNPs) of GPCRs. Even though RT-qPCR has become the standard method for quantification of gene expression, RT-qPCR is sensitive to RNA quality, assay design, normalisation approach and data analysis. This protocol is meant as a guide to RT-qPCR methodology with references to the best standard methods available at present.

  17. Species identification of Asini Corii Collas (donkey glue) by PCR amplification of cytochrome b gene.

    Science.gov (United States)

    Kumeta, Yukie; Maruyama, Takuro; Asama, Hiroshi; Yamamoto, Yutaka; Hakamatsuka, Takashi; Goda, Yukihiro

    2014-01-01

    Asini Corii Collas (ACC; donkey glue) is a crude drug used to promote hematopoiesis and arrest bleeding. Because adulteration of the drug with substances from other animals such as horses, cattle, and pigs has been found, we examined PCR methods based on the sequence of the cytochrome b gene for source species identification. Two strategies for extracting DNA from ACC were compared, and the ion-exchange resin procedure was revealed to be more suitable than the silica-based one. Using DNA extracted from ACC by the ion-exchange resin procedure, PCR methods for species-specific detection of donkey, horse, cattle, and pig substances were established. When these species-specific PCR methods were applied to ACC, amplicons were obtained only by the donkey-specific PCR. Cattle-specific PCR detected as little as 0.1% admixture of cattle glue in the ACC. These results suggest that the species-specific PCR methods established in this study would be useful for simple and easy detection of adulteration of ACC.

  18. Detection of Streptococcus dysgalactiae subsp. equisimilis in equine nasopharyngeal swabs by PCR.

    Science.gov (United States)

    Preziuso, Silvia; Laus, Fulvio; Tejeda, Aurora Romero; Valente, Carlo; Cuteri, Vincenzo

    2010-03-01

    Streptococcus (S.) dysgalactiae subsp. equisimilis is responsible for severe diseases in humans, including primary bacteraemia, pneumonia, endocarditis, and toxic shock syndrome. Infection in some animal species can also occur, although a few studies have looked into cross-species infectivity. In horses, S. equisimilis is generally considered infrequent or opportunistic, but has recently been isolated from cases of strangles-like disease. Rapid and sensitive diagnostic techniques could enable epidemiological studies and effective investigation of outbreaks involving these bacteria. In this study, PCR protocols previously described in cattle and in humans to detect the species S. dysgalactiae and the subspecies equisimilis were evaluated to detect specific sequences in equine samples. For this purpose, 99 monolateral nasal swabs were collected from horses from stud farms with a history of S. equisimilis infection and were tested blindly by bacteriological isolation and by single and duplex PCR. DNA for PCR was extracted both from the colonies grown on agar media and from enrichment broth aliquots after incubation with nasal swab samples. S. equisimilis was identified by bacteriological isolation in 23 out of 99 swab samples, and PCR assays on these colonies were fully concordant with bacteriological identification (kappa statistic = 1.00). In addition, PCR of the enrichment broth aliquots confirmed the bacteriological results and detected S. equisimilis in 6 samples more than the bacteriological examination (kappa statistic = 0.84). The PCR protocols appeared to be reliable for the rapid identification of S. equisimilis in equine nasal swab samples, and could be useful for microbiological diagnosis.

  19. Simple Animations with Excels

    Science.gov (United States)

    Blickensderfer, Roger

    2010-01-01

    In recent years there has been a rapid expansion in the use of animated drawings for teaching physics. The benefits to the students are obvious. Rather than looking at still pictures in a textbook, they can observe a physical event and see how it plays out over time.

  20. Transgenic Farm Animals

    Science.gov (United States)

    The development of recombinant DNA technology has enabled scientists to isolate single genes, analyze and modify their nucleotide structure(s), make copies of these isolated genes, and insert copies of these genes into the genome of plants and animals. The transgenic technology of adding genes to li...

  1. Animals that Live Longest

    Institute of Scientific and Technical Information of China (English)

    饶扬志

    2000-01-01

    Reptiles(爬行类) are animals that live longest. The turtle's(海龟)long life is legendary(传奇的), no one has ever been able to calculate the exact age of the turtle, and for good reason, tortoises live a lot longer than humans do.

  2. In and Out (Animation)

    Science.gov (United States)

    2004-01-01

    This animation links two images taken by the front hazard avoidance camera on the Mars Exploration Rover Spirit. The rover is stowing and unstowing its robotic arm, or instrument deployment device. The device is designed to hold and maneuver the various instruments on board that will help scientists get up-close and personal with martian rocks and soil.

  3. Do Animals Have Memes?

    NARCIS (Netherlands)

    Reader, S.M.; Laland, K.N.

    1999-01-01

    Imitation has been put forward as a defining feature of memetic transmission. Since there is currently poor evidence for imitation in non-human animals, such definitions have been interpreted as restricting meme theory to the study of human behaviour patterns and birdsong. We believe this is a mista

  4. Farm animal welfare

    DEFF Research Database (Denmark)

    Sandøe, Peter; Christiansen, Stine Billeschou; Appleby, M. C.

    2003-01-01

    An experimental survey was undertaken to explore the links between the characteristics of a moral issue, the degree of moral intensity/moral imperative associated with the issue (Jones, 1991), and people’s stated willingness to pay (wtp) for policy to address the issue. Two farm animal welfare...

  5. Modelling Farm Animal Welfare.

    Science.gov (United States)

    Collins, Lisa M; Part, Chérie E

    2013-05-16

    The use of models in the life sciences has greatly expanded in scope and advanced in technique in recent decades. However, the range, type and complexity of models used in farm animal welfare is comparatively poor, despite the great scope for use of modeling in this field of research. In this paper, we review the different modeling approaches used in farm animal welfare science to date, discussing the types of questions they have been used to answer, the merits and problems associated with the method, and possible future applications of each technique. We find that the most frequently published types of model used in farm animal welfare are conceptual and assessment models; two types of model that are frequently (though not exclusively) based on expert opinion. Simulation, optimization, scenario, and systems modeling approaches are rarer in animal welfare, despite being commonly used in other related fields. Finally, common issues such as a lack of quantitative data to parameterize models, and model selection and validation are discussed throughout the review, with possible solutions and alternative approaches suggested.

  6. Modelling Farm Animal Welfare

    Directory of Open Access Journals (Sweden)

    Chérie E. Part

    2013-05-01

    Full Text Available The use of models in the life sciences has greatly expanded in scope and advanced in technique in recent decades. However, the range, type and complexity of models used in farm animal welfare is comparatively poor, despite the great scope for use of modeling in this field of research. In this paper, we review the different modeling approaches used in farm animal welfare science to date, discussing the types of questions they have been used to answer, the merits and problems associated with the method, and possible future applications of each technique. We find that the most frequently published types of model used in farm animal welfare are conceptual and assessment models; two types of model that are frequently (though not exclusively based on expert opinion. Simulation, optimization, scenario, and systems modeling approaches are rarer in animal welfare, despite being commonly used in other related fields. Finally, common issues such as a lack of quantitative data to parameterize models, and model selection and validation are discussed throughout the review, with possible solutions and alternative approaches suggested.

  7. Animal brucellosis in Egypt.

    Science.gov (United States)

    Wareth, Gamal; Hikal, Ahmed; Refai, Mohamed; Melzer, Falk; Roesler, Uwe; Neubauer, Heinrich

    2014-11-13

    Brucellosis is a highly contagious zoonosis that affects the public health and economic performance of endemic as well as non-endemic countries. In developing nations, brucellosis is often a very common but neglected disease. The purpose of this review is to provide insight about brucellosis in animal populations in Egypt and help to understand the situation from 1986 to 2013. A total of 67 national and international scientific publications on serological investigations, isolation, and biotyping studies from 1986 to 2013 were reviewed to verify the current status of brucellosis in animal populations in Egypt. Serological investigations within the national surveillance program give indirect proof for the presence of brucellosis in cattle, buffaloes, sheep, goats, and camels in Egypt. Serologic testing for brucellosis is a well-established procedure in Egypt, but most of the corresponding studies do not follow the scientific standards. B. melitensis biovar (bv) 3, B. abortus bv 1, and B. suis bv 1 have been isolated from farm animals and Nile catfish. Brucellosis is prevalent nationwide in many farm animal species. There is an obvious discrepancy between official seroprevalence data and data from scientific publications. The need for a nationwide survey to genotype circulating Brucellae is obvious. The epidemiologic situation of brucellosis in Egypt is unresolved and needs clarification.

  8. Cytogenetics in animal production

    Directory of Open Access Journals (Sweden)

    L. Iannuzzi

    2010-04-01

    Full Text Available Cytogenetics applied to domestic animals is a useful biotechnology to be applied in the genetic improvement of livestock. Indeed, it can be used to select reproducers free chromosome abnormalities which are responsible for abnormal body conformation (aneuploidy, lower fertility (balanced chromosome abnormalities or sterility (sex chromosome abnormalities. Cytogenetics may also be applied to assess environmental pollution by studying animals living in hazardous areas and using them as biological indicators (sentinels. Chromosomes also represent optimal biological structures to study the evolution among related (bovids and unrelated (bovidshumans species, especially using comparative FISH-mapping which is one of the most powerful tools to establish the correct order of loci along chromosomes. These comparisons allow us to transfer useful information from richer genomes (human to those of domestic animals. Moreover, the use of specific molecular markers and the FISH-technique on both mitotic and extended (fiber-FISH chromosomes, has heralded a new era of cytogenetics, allowing swift extension of genetic physical maps, better anchoring of both linkage and RH-maps to specific chromosome regions, and use in a variety of applications (clinical cases, embryo and sperm analyses, evolution. In this study a brief review of these fields of the animal cytogenetics is presented.

  9. Animation of Antimicrobial Resistance

    Medline Plus

    Full Text Available ... links HHS U.S. Department of Health and Human Services U.S. Food and Drug Administration A to Z Index Follow FDA En Español Search FDA Submit search Popular Content Home Food Drugs Medical Devices Radiation-Emitting Products Vaccines, Blood & Biologics Animal & Veterinary Cosmetics Tobacco Products ...

  10. Laboratory animal allergy.

    NARCIS (Netherlands)

    Hollander, A.

    1997-01-01

    The main objective of the study presented in this thesis was to estimate the prevalence rate of laboratory animal allergy and to determine its association with risk factors, like allergen exposure level, atopy, gender and other host factors. A cross-sectional survey was undertaken among 540 workers

  11. Pathological anxiety in animals

    NARCIS (Netherlands)

    Ohl, F.; Arndt, S.S.; Staay, van der F.J.

    2008-01-01

    selective breeding programmes in domestic and laboratory animals generally focus on physiological and/or anatomical characteristics. However, selection may have an (unintended) impact on other characteristics and may lead to dysfunctional behaviour that can affect biological functioning and, as a co

  12. Can Animals Think?

    Institute of Scientific and Technical Information of China (English)

    1994-01-01

    For centuries, philosophers argued that thinking and language sepa-rate humans from other species. The lesser creatures, Rene Descartes con-tended in I637, are little more than automatons, sleepwalking through lifewithout a mote of self-awareness. Later, scientists had reason to be skep-tical of claims concerning animal intelligence. At the turn of the century,

  13. Specific PCR and real-time PCR assays for detection and quantitation of 'Candidatus Phytoplasma phoenicium'.

    Science.gov (United States)

    Jawhari, Maan; Abrahamian, Peter; Sater, Ali Abdel; Sobh, Hana; Tawidian, Patil; Abou-Jawdah, Yusuf

    2015-02-01

    Almond witches' broom (AlmWB) is a fast-spreading lethal disease of almond, peach and nectarine associated with 'Candidatus Phytoplasma phoenicium'. The development of PCR and quantitative real-time PCR (qPCR) assays for the sensitive and specific detection of the phytoplasma is of prime importance for early detection of 'Ca. P. phoenicium' and for epidemiological studies. The developed qPCR assay herein uses a TaqMan(®) probe labeled with Black Hole Quencher Plus. The specificity of the PCR and that of the qPCR detection protocols were tested on 17 phytoplasma isolates belonging to 11 phytoplasma 16S rRNA groups, on samples of almond, peach, nectarine, native plants and insects infected or uninfected with the phytoplasma. The developed assays showed high specificity against 'Ca. P. phoenicium' and no cross-reactivity against any other phytoplasma, plant or insect tested. The sensitivity of the developed PCR and qPCR assays was similar to the conventional nested PCR protocol using universal primers. The qPCR assay was further validated by quantitating AlmWB phytoplasma in different hosts, plant parts and potential insect vectors. The highest titers of 'Ca. P. phoenicium' were detected in the phloem tissues of stems and roots of almond and nectarine trees, where they averaged from 10(5) to 10(6) genomic units per nanogram of host DNA (GU/ng of DNA). The newly developed PCR and qPCR protocols are reliable, specific and sensitive methods that are easily applicable to high-throughput diagnosis of AlmWB in plants and insects and can be used for surveys of potential vectors and alternative hosts.

  14. Reatividade animal Confinement reactivity

    Directory of Open Access Journals (Sweden)

    Walsiara Estanislau Maffei

    2009-07-01

    Full Text Available A reatividade é definida como a reação do animal quando contido num ambiente de contenção móvel. Ela é quantificada por meio do teste de reatividade animal em ambiente de contenção móvel - REATEST®. Este teste consiste num dispositivo eletrônico acoplado à balança e num software específico. O dispositivo capta a movimentação que o animal provoca na balança, durante 20 segundos e a envia para o software que a processa determinando a reatividade do animal numa escala contínua de pontos. Pontuações maiores são de animais mais reativos (mais agressivo. A reatividade foi criada com os objetivos de solucionar os problemas até então existentes na seleção para temperamento e de permitir estimação de parâmetros genéticos mais confiáveis. Ela é uma característica objetiva que tem grande variabilidade fenotípica e é de quantificação rápida, fácil e segura, além de poder ser quantificada em qualquer tipo de balança, o que permite maior aplicabilidade. Ela não interfere nas práticas de manejo das fazendas porque é quantificada no momento da pesagem dos animais. Sua herdabilidade na raça Nelore é de 0,39 ao ano e 0,23 ao sobreano e suas correlações genéticas com ganho de peso diário são de -0,28 do nascimento até desmama e de -0,49 do desmame até ano. Já suas correlações genéticas com desenvolvimento do perímetro escrotal do ano ao sobreano variam de -0,25 e -0,41.The confinement reactivity (CR has been used as a measure of temperament in Brazil and it is defined as the animal reaction when contained in the scale. It is quantified through the animal reactivity test - REATEST®. This test consists of an electronic device coupled to the scale and of specific software. The device captures the movement that the animal provokes in the scale, during 20 seconds and sends it for the software that processes this movement and determines the animal CR in a continuous scale of points. Higher punctuations belong to

  15. A method for amplification of unknown flanking sequences based on touchdown PCR and suppression-PCR.

    Science.gov (United States)

    Gao, Song; He, Dan; Li, Guangquan; Zhang, Yanhua; Lv, Huiying; Wang, Li

    2016-09-15

    Thermal asymmetric staggered PCR is the most widely used technique to obtain the flanking sequences. However, it has some limitations, including a low rate of positivity, and complex operation. In this study, a improved method of it was made based on suppression-PCR and touchdown PCR. The PCR fragment obtained by the amplification was used directly for sequencing after gel purification. Using this improved method, the positive rate of amplified flanking sequences of the ATMT mutants reached 99%. In addition, the time from DNA extraction to flanking sequence analysis was shortened to 2 days with about 6 dollars each sample.

  16. Signal and noise in bridging PCR

    Directory of Open Access Journals (Sweden)

    Thaler David S

    2002-07-01

    Full Text Available Abstract Background In a variant of the standard PCR reaction termed bridging, or jumping, PCR the primer-bound sequences are originally on separate template molecules. Bridging can occur if, and only if, the templates contain a region of sequence similarity. A 3' end of synthesis in one round of synthesis that terminates in this region of similarity can prime on the other. In principle, Bridging PCR (BPCR can detect a subpopulation of one template that terminates synthesis in the region of sequence shared by the other template. This study considers the sensitivity and noise of BPCR as a quantitative assay for backbone interruptions. Bridging synthesis is also important to some methods for computing with DNA. Results In this study, BPCR was tested over a 328 base pair segment of the E. coli lac operon and a signal to noise ratio (S/N of approximately 10 was obtained under normal PCR conditions with Taq polymerase. With special precautions in the case of Taq or by using the Stoffel fragment the S/N was improved to 100, i.e. 1 part of cut input DNA yielded the same output as 100 parts of intact input DNA. Conclusions In the E. coli lac operator region studied here, depending on details of protocol, between 3 and 30% per kilobase of final PCR product resulted from bridging. Other systems are expected to differ in the proportion of product that is bridged consequent to PCR protocol and the sequence analyzed. In many cases physical bridging during PCR will have no informational consequence because the bridged templates are of identical sequence, but in a number of special cases bridging creates, or, destroys, information.

  17. Determinación de la estructura poblacional de litopenaeus vannamei mediante issrs a lo largo de la costa ecuatoriana

    OpenAIRE

    Fresneda Rodríguez, Adriana

    2003-01-01

    Determinación de la estructura poblacional de Litopenaeus vannamei mediante ISSRs a lo largo de la costa ecuatoriana Muestras de Litopenaeus vannamei fueron colectadas en siete localidades a lo largo de la costa ecuatoriana desde Esmeraldas hasta Machala.

  18. Fostering Kinship with Animals: Animal Portraiture in Humane Education

    Science.gov (United States)

    Kalof, Linda; Zammit-Lucia, Joe; Bell, Jessica; Granter, Gina

    2016-01-01

    Visual depictions of animals can alter human perceptions of, emotional responses to, and attitudes toward animals. Our study addressed the potential of a slideshow designed to activate emotional responses to animals to foster feelings of kinship with them. The personal meaning map measured changes in perceptions of animals. The participants were…

  19. Nested PCR for specific diagnosis of Taenia solium taeniasis.

    Science.gov (United States)

    Mayta, Holger; Gilman, Robert H; Prendergast, Emily; Castillo, Janeth P; Tinoco, Yeny O; Garcia, Hector H; Gonzalez, Armando E; Sterling, Charles R

    2008-01-01

    Taeniasis due to Taenia solium is a disease with important public health consequences, since the larval stage is not exclusive to the animal intermediate, the pig, but also infects humans, causing neurocysticercosis. Early diagnosis and treatment of T. solium tapeworm carriers is important to prevent human cysticercosis. Current diagnosis based on microscopic observation of eggs lacks both sensitivity and specificity. In the present study, a nested-PCR assay targeting the Tso31 gene was developed for the specific diagnosis of taeniasis due to T. solium. Initial specificity and sensitivity testing was performed using stored known T. solium-positive and -negative samples. The assay was further analyzed under field conditions by conducting a case-control study of pretreatment stool samples collected from a population in an area of endemicity. Using the archived samples, the assay showed 97% (31/32) sensitivity and 100% (123/123) specificity. Under field conditions, the assay had 100% sensitivity and specificity using microscopy/enzyme-linked immunosorbent assay coproantigen testing as the gold standards. The Tso31 nested PCR described here might be a useful tool for the early diagnosis and prevention of taeniasis/cysticercosis.

  20. SIMULACIÓN DEL COLAPSO EN TÚNELES DOVELADOS MEDIANTE LA FORMACIÓN DE ARTICULACIONES EN VIGAS

    OpenAIRE

    Enrique Tenorio Montero; Gelacio Juárez Luna

    2013-01-01

    Se estudia el mecanismo de colapso en túneles construidos con dovelas mediante el método los de elementos finitos. Los túneles se discretizaron mediante elementos finitos viga, resortes longitudinales y rotacionales, estos últimos tienen la capacidad de simular el desarrollo de articulaciones y el comportamiento de las juntas longitudinales, considerando un comportamiento constitutivo momento-rotación de las dovelas. En los apoyos se colocaron resortes axiales elásticos que representan la rig...

  1. Capacidad discriminatoria y concordancia entre el ELISA-F29 y la PCR en individuos con infección por T. cruzi / Discriminatory Power and Concordance between ELISA-F29 and PCR in Individuals with Infection due to T.cruzi / Capacidade discriminatória e acordo entre o ELISA-F29 e a PCR em indivíduos infectados com T. cruzi

    OpenAIRE

    Edgar David Gómez-Laitón, MD.; Laura Andrea Polo-Ardila, Enf.; Yeny Zulay Castellanos-Domínguez, Bact., Esp., MsC; Víctor Mauricio Herrera-Galindo, MD., MSc., PhD.; Juan Carlos Villar-Centeno, MD., Esp., MSc., PhD

    2015-01-01

    Introducción: El diagnóstico de la infección por Trypanosoma cruzi (T. cruzi) se realiza rutinariamente mediante pruebas serológicas mientras que el empleo de métodos moleculares se encuentra aún en proceso de estandarización. Objetivo: Evaluar la capacidad discriminatoria y concordancia entre una prueba serológica y una molecular para determinar la infección por T. cruzi. Métodos: Se realizó Reacción en Cadena de la Polimerasa (PCR) y la prueba de ELISA-F29 en 95 muestras de participantes d...

  2. PCR+ In Diesel Fuels and Emissions Research

    Energy Technology Data Exchange (ETDEWEB)

    McAdams, H.T.

    2002-04-15

    In past work for the U.S. Department of Energy (DOE) and Oak Ridge National Laboratory (ORNL), PCR+ was developed as an alternative methodology for building statistical models. PCR+ is an extension of Principal Components Regression (PCR), in which the eigenvectors resulting from Principal Components Analysis (PCA) are used as predictor variables in regression analysis. The work was motivated by the observation that most heavy-duty diesel (HDD) engine research was conducted with test fuels that had been ''concocted'' in the laboratory to vary selected fuel properties in isolation from each other. This approach departs markedly from the real world, where the reformulation of diesel fuels for almost any purpose leads to changes in a number of interrelated properties. In this work, we present new information regarding the problems encountered in the conventional approach to model-building and how the PCR+ method can be used to improve research on the relationship between fuel characteristics and engine emissions. We also discuss how PCR+ can be applied to a variety of other research problems related to diesel fuels.

  3. 18S Ribosomal RNA Evaluation as Preanalytical Quality Control for Animal DNA

    Directory of Open Access Journals (Sweden)

    Cory Ann Leonard

    2016-01-01

    Full Text Available The 18S ribosomal RNA (rRNA gene is present in all eukaryotic cells. In this study, we evaluated the use of this gene to verify the presence of PCR-amplifiable host (animal DNA as an indicator of sufficient sample quality for quantitative real-time PCR (qPCR analysis. We compared (i samples from various animal species, tissues, and sample types, including swabs; (ii multiple DNA extraction methods; and (iii both fresh and formalin-fixed paraffin-embedded (FFPE samples. Results showed that 18S ribosomal RNA gene amplification was possible from all tissue samples evaluated, including avian, reptile, and FFPE samples and most swab samples. A single swine rectal swab, which showed sufficient DNA quantity and the demonstrated lack of PCR inhibitors, nonetheless was negative by 18S qPCR. Such a sample specifically illustrates the improvement of determination of sample integrity afforded by inclusion of 18S rRNA gene qPCR analysis in addition to spectrophotometric analysis and the use of internal controls for PCR inhibition. Other possible applications for the described 18S rRNA qPCR are preselection of optimal tissue specimens for studies or preliminary screening of archived samples prior to acceptance for biobanking projects.

  4. Direct chromatin PCR (DC-PCR: hypotonic conditions allow differentiation of chromatin states during thermal cycling.

    Directory of Open Access Journals (Sweden)

    Sergei Vatolin

    Full Text Available Current methods to study chromatin configuration are not well suited for high throughput drug screening since they require large cell numbers and multiple experimental steps that include centrifugation for isolation of nuclei or DNA. Here we show that site specific chromatin analysis can be achieved in one step by simply performing direct chromatin PCR (DC-PCR on cells. The basic underlying observation was that standard hypotonic PCR buffers prevent global cellular chromatin solubilization during thermal cycling while more loosely organized chromatin can be amplified. Despite repeated heating to >90 °C, 41 of 61 tested 5' sequences of silenced genes (CDKN2A, PU.1, IRF4, FOSB, CD34 were not amplifiable while 47 could be amplified from expressing cells. Two gene regions (IRF4, FOSB even required pre-heating of cells in isotonic media to allow this differentiation; otherwise none of 19 assayed sequences yielded PCR products. Cells with baseline expression or epigenetic reactivation gave similar DC-PCR results. Silencing during differentiation of CD34 positive cord blood cells closed respective chromatin while treatment of myeloma cells with an IRF4 transcriptional inhibitor opened a site to DC-PCR that was occupied by RNA polymerase II and NFκB as determined by ChIP. Translation into real-time PCR can not be achieved with commercial real-time PCR buffers which potently open chromatin, but even with simple ethidium bromide addition to standard PCR mastermix we were able to identify hits in small molecules screens that suppressed IRF4 expression or reactivated CDKN2A in myeloma cells using densitometry or visual inspection of PCR plates under UV light. While need in drug development inspired this work, application to genome-wide analysis appears feasible using phi29 for selective amplification of open cellular chromatin followed by library construction from supernatants since such supernatants yielded similar results as gene specific DC-PCR.

  5. Animal welfare and use of silkworm as a model animal.

    Science.gov (United States)

    Sekimizu, N; Paudel, A; Hamamoto, H

    2012-08-01

    Sacrificing model animals is required for developing effective drugs before being used in human beings. In Japan today, at least 4,210,000 mice and other mammals are sacrificed to a total of 6,140,000 per year for the purpose of medical studies. All the animals treated in Japan, including test animals, are managed under control of "Act on Welfare and Management of Animals". Under the principle of this Act, no person shall kill, injure, or inflict cruelty on animals without due cause. "Animal" addressed in the Act can be defined as a "vertebrate animal". If we can make use of invertebrate animals in testing instead of vertebrate ones, that would be a remarkable solution for the issue of animal welfare. Furthermore, there are numerous advantages of using invertebrate animal models: less space and small equipment are enough for taking care of a large number of animals and thus are cost-effective, they can be easily handled, and many biological processes and genes are conserved between mammals and invertebrates. Today, many invertebrates have been used as animal models, but silkworms have many beneficial traits compared to mammals as well as other insects. In a Genome Pharmaceutical Institute's study, we were able to achieve a lot making use of silkworms as model animals. We would like to suggest that pharmaceutical companies and institutes consider the use of the silkworm as a model animal which is efficacious both for financial value by cost cutting and ethical aspects in animals' welfare.

  6. Fur animal botulism hazard due to feed.

    Science.gov (United States)

    Myllykoski, J; Lindström, M; Bekema, E; Pölönen, I; Korkeala, H

    2011-06-01

    To assess the botulism hazard in fur animal feed production, 236 fur animal feed components and feed samples were analysed for Clostridium botulinum by detecting BoNT-encoding genes (botA, botB, botC, botE or botF) by PCR and for sulphite-reducing clostridia (SRC) by iron sulphite agar. The quality of the hazard analysis of critical control points (HACCP) -based in-house control system (IHCS) was evaluated with respect to botulism risk in feed plants (n=32). The overall prevalence of C. botulinum was 13% in different feed components and 5% in feed. The estimated MPN count of C. botulinum in feed components was 6.4 × 10(3)/kg at the highest and was shown to poorly correlate with SRC count. The critical control points in IHCSs were variable, and control limits were improperly set in most feed-producing plants. C. botulinum possesses a persistent safety hazard for fur animals by feed production, and control practices should be reassessed.

  7. Comparison of a quantitative real-time polymerase chain reaction (qPCR) with conventional PCR, bacterial culture and ELISA for detection of Mycobacterium avium subsp. paratuberculosis infection in sheep showing pathology of Johne's disease.

    Science.gov (United States)

    Sonawane, Ganesh G; Tripathi, Bhupendra N

    2013-12-01

    A quantitative real-time PCR (qPCR) assay employing IS900 gene specific primers of Mycobacterium avium subsp. parartuberculosis (MAP) was compared with conventional PCR, bacterial culture and enzyme-linked immunosorbent assay in 38 sheep showing granulomatous enteritis and lymphadenitis with and without demonstration of acid-fast bacilli (AFB). The lesions were classified as multibacillary (MB) (n = 23), which had diffuse granulomatous lesions with abundant AFB, and paucibacillary (PB) (n = 15), which had focal or multifocal granulomatous lesions with few or no AFB. In the multibacillary group (MB), IS900 PCR detected 19 (82.6%), and qPCR detected all 23 (100%) sheep positive for MAP in the intestine and lymph node tissues. In the paucibacillary group (PB), IS900 PCR detected 2 (13.3%), and qPCR detected all 15 (100%) sheep positive for MAP in tissues. When results of both groups were taken together, IS900 PCR detected 21(55.2%), and qPCR detected all 38 (100%) animals positive for MAP genome either in the intestine or lymph node tissues. On Herrold egg yolk medium, tissues of 14 (60.9%) MB and 5 (33.3%) PB sheep were found to be positive for MAP. Out of 27 sheep (PB = 8, MB = 19) tested by an ELISA, 21 (77.7%) were found to be positive for MAP antibody, of which 25% (2/8) and 100% (19/19) sheep were from PB and MB sheep, respectively. Based on the results of the present study, it was concluded that qPCR was a highly sensitive test in comparison to conventional PCR, ELISA and bacterial culture for the diagnosis of paratuberculosis on infected tissues especially from paucibacillary sheep.

  8. Animal bites - self-care

    Science.gov (United States)

    Bites - animals - self-care ... Most animal bites come from pets. Dog bites are common and most often happen to children. Cat bites are ... which can cause deeper puncture wounds. Most other animal bites are caused by stray or wild animals, ...

  9. Professional perspectives on animal hoarding.

    OpenAIRE

    Burniston, Francesca A.

    2016-01-01

    Objectives Although theoretical conceptualisations of animal hoarding have been published, few empirical studies have been conducted. The current study investigated animal hoarding from the perspectives of professionals who come into contact with people who hoard animals through their employment in various capacities, primarily in animal welfare. Design A qualitative research design was employed using inductive thematic analysis. Methods Twelve professionals who had experienc...

  10. Analysis of Animal Metaphorical Expression

    Institute of Scientific and Technical Information of China (English)

    姜晴川

    2016-01-01

    Animal metaphor, as a kind of metaphor, refers to a cognitive process in which some aspects of human beings are understood or experienced through the aspects of animals. The meanings of animal metaphor are based on people's experience, cultural background, custom and the ways of thinking. Animal metaphorical expression is an important part of human's language expressions and communication.

  11. Animals:Country symbols

    Institute of Scientific and Technical Information of China (English)

    周明

    2005-01-01

    A nim als have always been used to represent cer-tain hum an characteristics.Countries also use anim alsas sym bols.From eagles to lions,m any countries usean anim al to show its national spirit and character.1.U S:T he bald eagleThe im age of an eagle is on the U SPresident’s flag,and on the one-dollarbill.The bald eagle is a large,pow erful,brow n bird with a white head and tail.The term“bald”does not m ean that thisbird lacks feathers.Instead,it com es fromthe old word piebald,that m eans,“m arked w ith ...

  12. Animal Gaits and Symmetry

    Science.gov (United States)

    Golubitsky, Martin

    2012-04-01

    Many gaits of four-legged animals are described by symmetry. For example, when a horse paces it moves both left legs in unison and then both right legs and so on. The motion is described by two symmetries: Interchange front and back legs, and swap left and right legs with a half-period phase shift. Biologists postulate the existence of a central pattern generator (CPG) in the neuronal system that sends periodic signals to the legs. CPGs can be thought of as electrical circuits that produce periodic signals and can be modeled by systems with symmetry. In this lecture we discuss animal gaits; use gait symmetries to construct a simplest CPG architecture that naturally produces quadrupedal gait rhythms; and make several testable predictions about gaits.

  13. Spectral Animation Compression

    Institute of Scientific and Technical Information of China (English)

    Chao Wang; Yang Liu; Xiaohu Guo; Zichun Zhong; Binh Le; Zhigang Deng

    2015-01-01

    This paper presents a spectral approach to compress dynamic animation consisting of a sequence of homeomor-phic manifold meshes. Our new approach directly compresses the field of deformation gradient defined on the surface mesh, by decomposing it into rigid-body motion (rotation) and non-rigid-body deformation (stretching) through polar decompo-sition. It is known that the rotation group has the algebraic topology of 3D ring, which is different from other operations like stretching. Thus we compress these two groups separately, by using Manifold Harmonics Transform to drop out their high-frequency details. Our experimental result shows that the proposed method achieves a good balance between the reconstruction quality and the compression ratio. We compare our results quantitatively with other existing approaches on animation compression, using standard measurement criteria.

  14. Advances in Animal Cognition.

    Science.gov (United States)

    Vonk, Jennifer

    2016-11-30

    This editorial endorses a diverse approach to the study of animal cognition and emphasizes the theoretical and applied gains that can be made by embracing this approach. This diversity emerges from cross-talk among scientists trained in a variety of backgrounds and theoretical approaches, who study a variety of topics with a range of species. By shifting from an anthropocentric focus on humans and our closest living relatives, and the historic reliance on the lab rat or pigeon, modern students of animal cognition have uncovered many fascinating facets of cognition in species ranging from insects to carnivores. Diversity in both topic and species of study will allow researchers to better understand the complex evolutionary forces giving rise to widely shared and unique cognitive processes. Furthermore, this increased understanding will translate into more effective strategies for managing wild and captive populations of nonhuman species.

  15. Phoenix Animation Looking North

    Science.gov (United States)

    2008-01-01

    [figure removed for brevity, see original site] Click on image for animation This animation is a series of images, taken by NASA's Phoenix Mars Lander's Surface Stereo Imager, combined into a panoramic view looking north from the lander. The area depicted is beyond the immediate workspace of the lander and shows a system of polygons and troughs that connect with the ones Phoenix will be investigating in depth. The images were taken on sol 14 (June 8, 2008) or the 14th Martian day after landing. The Phoenix Mission is led by the University of Arizona, Tucson, on behalf of NASA. Project management of the mission is by NASA's Jet Propulsion Laboratory, Pasadena, Calif. Spacecraft development is by Lockheed Martin Space Systems, Denver.

  16. Phoenix Work Area Animation

    Science.gov (United States)

    2008-01-01

    [figure removed for brevity, see original site] Click on image for animation This animation from Sol 1 shows a mosaic of the Phoenix digging area in the Martian terrain. Phoenix scientists are very pleased with this view as the terrain features few rocks an optimal place for digging. The mast of the camera looks disjointed because the photos that comprise this mosaic were taken at different times of day. This video also show some of the lander's instrumentation. The Phoenix Mission is led by the University of Arizona, Tucson, on behalf of NASA. Project management of the mission is by NASA's Jet Propulsion Laboratory, Pasadena, Calif. Spacecraft development is by Lockheed Martin Space Systems, Denver.

  17. Animal Capture Agents

    Science.gov (United States)

    1990-01-01

    agents and delivery systems reviewed . Questionnaires were sent to 137 Air Force bases to obtain information about the chemical agents and delivery systems...used by animal control personnel. A literature review included chemical agents, delivery methods, toxicity information and emergency procedures from...34-like agent. Users should familiarize themselves with catatonia in general and particularly that its successful use as an immobilizer doesn’t necessarily

  18. Pain in aquatic animals.

    Science.gov (United States)

    Sneddon, Lynne U

    2015-04-01

    Recent developments in the study of pain in animals have demonstrated the potential for pain perception in a variety of wholly aquatic species such as molluscs, crustaceans and fish. This allows us to gain insight into how the ecological pressures and differential life history of living in a watery medium can yield novel data that inform the comparative physiology and evolution of pain. Nociception is the simple detection of potentially painful stimuli usually accompanied by a reflex withdrawal response, and nociceptors have been found in aquatic invertebrates such as the sea slug Aplysia. It would seem adaptive to have a warning system that allows animals to avoid life-threatening injury, yet debate does still continue over the capacity for non-mammalian species to experience the discomfort or suffering that is a key component of pain rather than a nociceptive reflex. Contemporary studies over the last 10 years have demonstrated that bony fish possess nociceptors that are similar to those in mammals; that they demonstrate pain-related changes in physiology and behaviour that are reduced by painkillers; that they exhibit higher brain activity when painfully stimulated; and that pain is more important than showing fear or anti-predator behaviour in bony fish. The neurophysiological basis of nociception or pain in fish is demonstrably similar to that in mammals. Pain perception in invertebrates is more controversial as they lack the vertebrate brain, yet recent research evidence confirms that there are behavioural changes in response to potentially painful events. This review will assess the field of pain perception in aquatic species, focusing on fish and selected invertebrate groups to interpret how research findings can inform our understanding of the physiology and evolution of pain. Further, if we accept these animals may be capable of experiencing the negative experience of pain, then the wider implications of human use of these animals should be considered.

  19. Instant Silverlight 5 animation

    CERN Document Server

    Polyak, Nick

    2013-01-01

    This book is written in simple, easy to understand format with lots of screenshots and step-by-step explanations. If you are a developer looking forward to create great user experience for your Silverlight applications with cool animations or create Silverlight banner ads, then this is the guide for you. It is assumed that the readers have some previous exposure to Silverlight or WPF.

  20. Animating the Carbon Cycle

    OpenAIRE

    2014-01-01

    Understanding the biogeochemical processes reg- ulating carbon cycling is central to mitigating atmospheric CO2 emissions. The role of living organisms has been accounted for, but the focus has traditionally been on contributions of plants and microbes. We develop the case that fully ‘‘animating’’ the carbon cycle requires broader consideration of the functional role of animals in mediating biogeochemical processes and quanti- fication of their effects on carbon storage and exchange among ter...

  1. Animal Models of Narcolepsy

    OpenAIRE

    Chen, Lichao; Brown, Ritchie E.; McKenna, James T.; McCarley, Robert W.

    2009-01-01

    Narcolepsy is a debilitating sleep disorder with excessive daytime sleepiness and cataplexy as its two major symptoms. Although this disease was first described about one century ago, an animal model was not available until the 1970s. With the establishment of the Stanford canine narcolepsy colony, researchers were able to conduct multiple neurochemical studies to explore the pathophysiology of this disease. It was concluded that there was an imbalance between monoaminergic and cholinergic sy...

  2. Diagnostic PCR tests for Microsporum audouinii, M. canis and Trichophyton infections

    DEFF Research Database (Denmark)

    Brillowska-Dabrowska, Anna; Swierkowska, Aleksandra; Lindhardt Saunte, Ditte Marie

    2010-01-01

    Since traditional diagnosis of dermatophyte infections is slow, we present a rapid new PCR test for detection of Trichophyton spp., Microsporum canis and M. audouinii infections. The performance of the test was evaluated with: 58 dermatophyte isolates; 10 yeast, mould and human DNA control samples......; 25 routine specimens from patients suspected of having dermatophytosis; 10 hair specimens from guinea pigs experimentally infected with M. canis; and two samples from un-infected control animals. DNA was prepared by a 10-min procedure from pure cultures as previously described. The 302 bp PCR product...... was obtained for 35/35 Trichophyton isolates (10 species included) and the 279 bp for 3/3 M. canis and 4/4 M. audouinii samples. None of the 2 E. floccosum, 11 M. gypseum, 3 M M. persicolor or 12 control samples (yeast, mould, human DNA) were positive with either of the two PCR tests. Among the patient...

  3. Detection of pork adulteration by highly-specific PCR assay of mitochondrial D-loop.

    Science.gov (United States)

    Karabasanavar, Nagappa S; Singh, S P; Kumar, Deepak; Shebannavar, Sunil N

    2014-02-15

    We describe a highly specific PCR assay for the authentic identification of pork. Accurate detection of tissues derived from pig (Sus scrofa) was accomplished by using newly designed primers targeting porcine mitochondrial displacement (D-loop) region that yielded an unique amplicon of 712 base pairs (bp). Possibility of cross-amplification was precluded by testing as many as 24 animal species (mammals, birds, rodent and fish). Suitability of PCR assay was confirmed in raw (n = 20), cooked (60, 80 and 100 °C), autoclaved (121 °C) and micro-oven processed pork. Sensitivity of detection of pork in other species meat using unique pig-specific PCR was established to be at 0.1%; limit of detection (LOD) of pig DNA was 10 pg (pico grams). The technique can be used for the authentication of raw, processed and adulterated pork and products under the circumstances of food adulteration related disputes or forensic detection of origin of pig species.

  4. History of animal bioacoustics

    Science.gov (United States)

    Popper, Arthur N.; Dooling, Robert J.

    2002-11-01

    The earliest studies on animal bioacoustics dealt largely with descriptions of sounds. Only later did they address issues of detection, discrimination, and categorization of complex communication sounds. This literature grew substantially over the last century. Using the Journal of the Acoustical Society of America as an example, the number of papers that fall broadly within the realm of animal sound production, communication, and hearing rose from two in the partial first decade of the journal in the 1930's, to 20 in the 1970's, to 92 in the first 2 years of this millennium. During this time there has been a great increase in the diversity of species studied, the sophistication of the methods used, and the complexity of the questions addressed. As an example, the first papers in JASA focused on a guinea pig and a bird. In contrast, since the year 2000 studies are often highly comparative and include fish, birds, dolphins, dogs, ants, crickets, and snapping shrimp. This paper on the history of animal bioacoustics will consider trends in work over the decades and discuss the formative work of a number of investigators who have spurred the field by making critical theoretical and experimental observations.

  5. Mathematical analysis of the real time array PCR (RTA PCR) process

    NARCIS (Netherlands)

    Dijksman J.F.; Pierik, A.

    2012-01-01

    Real Time Array PCR is a recently developed biochemical technique that measures amplification curves (like quantitative real time Polymerase Chain Reaction (qPCR)) of a multitude of different templates ina sample. It combines two different techniques to profit from theadvantages of both techniques,

  6. PCR melting profile (PCR MP - a new tool for differentiation of Candida albicans strains

    Directory of Open Access Journals (Sweden)

    Nowak Magdalena

    2009-11-01

    Full Text Available Abstract Background We have previously reported the use of PCR Melting Profile (PCR MP technique based on using low denaturation temperatures during ligation mediated PCR (LM PCR for bacterial strain differentiation. The aim of the current study was to evaluate this method for intra-species differentiation of Candida albicans strains. Methods In total 123 Candida albicans strains (including 7 reference, 11 clinical unrelated, and 105 isolates from patients of two hospitals in Poland were examined using three genotyping methods: PCR MP, macrorestriction analysis of the chromosomal DNA by pulsed-field gel electrophoresis (REA-PFGE and RAPD techniques. Results The genotyping results of the PCR MP were compared with results from REA-PFGE and RAPD techniques giving 27, 26 and 25 unique types, respectively. The results showed that the PCR MP technique has at least the same discriminatory power as REA-PFGE and RAPD. Conclusion Data presented here show for the first time the evaluation of PCR MP technique for candidial strains differentiation and we propose that this can be used as a relatively simple and cheap technique for epidemiological studies in short period of time in hospital.

  7. pcrEfficiency: a Web tool for PCR amplification efficiency prediction

    Directory of Open Access Journals (Sweden)

    Mallona Izaskun

    2011-10-01

    Full Text Available Abstract Background Relative calculation of differential gene expression in quantitative PCR reactions requires comparison between amplification experiments that include reference genes and genes under study. Ignoring the differences between their efficiencies may lead to miscalculation of gene expression even with the same starting amount of template. Although there are several tools performing PCR primer design, there is no tool available that predicts PCR efficiency for a given amplicon and primer pair. Results We have used a statistical approach based on 90 primer pair combinations amplifying templates from bacteria, yeast, plants and humans, ranging in size between 74 and 907 bp to identify the parameters that affect PCR efficiency. We developed a generalized additive model fitting the data and constructed an open source Web interface that allows the obtention of oligonucleotides optimized for PCR with predicted amplification efficiencies starting from a given sequence. Conclusions pcrEfficiency provides an easy-to-use web interface allowing the prediction of PCR efficiencies prior to web lab experiments thus easing quantitative real-time PCR set-up. A web-based service as well the source code are provided freely at http://srvgen.upct.es/efficiency.html under the GPL v2 license.

  8. Diagnosis of Cutaneous Leishmaniasis by Multiplex PCR

    Directory of Open Access Journals (Sweden)

    M Heiat

    2010-07-01

    Full Text Available Introduction: Annually, more than 14 million people are reported to be infected with Leishmaniasis all over the world. In Iran, this disease is seen in the form of cutaneous and visceral leishmaniasis, of which the cutaneous form is more wide spread. In recent years, cutaneous leishmaniaisis is diagnosed by PCR utilizing specific primers in order to amplify different parasite genes including ribosomal RNA genes, kinetoplast DNA or tandem repeating sequences. The aim of this research was to detect early stage cutaneous leishmaniasis using Multiplex-PCR technique. Methods: In this study, 67 samples were prepared from patients with cutaneous leishmaniasis. DNA was extracted with phenolchloroform. Each specimen was analyzed using two different pairs of PCR primers. The sensitivity of each PCR was optimized on pure Leishmania DNA prior to use for diagnosis. Two standard parasites L. major and L. tropica were used as positive control. Results: DNA amplification fragments were two 115 bp and 683 bp for AB and UL primers, respectively. The sensitivity of two primers was not equal for detection of L. major and L. tropica. The sensivity of PCR with AB primer was 35 cells, while that for UL primer was 40 cells. Conclusion: The results of this study indicate that PCR is a sensitive diagnostic assay for cutaneous leishmaniasis and could be employed as the new standard for routine diagnosis when species identification is not required. However, the ability to identify species is especially important in prognosis of the disease and in deciding appropriate therapy, especially in regions where more than one type of species and disease are seen by clinicians.

  9. The use of singleplex and nested PCR to detect Batrachochytrium dendrobatidis in free-living frogs

    Science.gov (United States)

    Coutinho, Selene Dall'Acqua; Burke, Julieta Catarina; de Paula, Catia Dejuste; Rodrigues, Miguel Trefaut; Catão-Dias, José Luiz

    2015-01-01

    Many microorganisms are able to cause diseases in amphibians, and in the past few years one of the most reported has been Batrachochytrium dendrobatidis. This fungus was first reported in Brazil in 2005; following this, other reports were made in specimens deposited in museum collections, captive and free-living frogs. The aim of this study was to compare singleplex and nested-PCR techniques to detect B. dendrobatidis in free-living and apparently healthy adult frogs from the Brazilian Atlantic Forest. The sample collection area was a protected government park, with no general entrance permitted and no management of the animals there. Swabs were taken from the skin of 107 animals without macroscopic lesions and they were maintained in ethanol p.a. Fungal DNA was extracted and identification of B. dendrobatidis was performed using singleplex and nested-PCR techniques, employing specific primers sequences. B. dendrobatidis was detected in 61/107 (57%) and 18/107 (17%) animals, respectively by nested and singleplex-PCR. Nested-PCR was statistically more sensible than the conventional for the detection of B. dendrobatidis (Chi-square = 37.1; α = 1%) and the agreement between both techniques was considered just fair (Kappa = 0.27). The high prevalence obtained confirms that these fungi occur in free-living frogs from the Brazilian Atlantic Forest with no macroscopic lesions, characterizing the state of asymptomatic carrier. We concluded that the nested-PCR technique, due to its ease of execution and reproducibility, can be recommended as one of the alternatives in epidemiological surveys to detect B. dendrobatidis in healthy free-living frog populations. PMID:26273273

  10. The use of singleplex and nested PCR to detect Batrachochytrium dendrobatidis in free-living frogs

    Directory of Open Access Journals (Sweden)

    Selene Dall'Acqua Coutinho

    2015-06-01

    Full Text Available Many microorganisms are able to cause diseases in amphibians, and in the past few years one of the most reported has been Batrachochytrium dendrobatidis. This fungus was first reported in Brazil in 2005; following this, other reports were made in specimens deposited in museum collections, captive and free-living frogs. The aim of this study was to compare singleplex and nested-PCR techniques to detect B. dendrobatidis in free-living and apparently healthy adult frogs from the Brazilian Atlantic Forest. The sample collection area was a protected government park, with no general entrance permitted and no management of the animals there. Swabs were taken from the skin of 107 animals without macroscopic lesions and they were maintained in ethanol p.a. Fungal DNA was extracted and identification of B. dendrobatidis was performed using singleplex and nested-PCR techniques, employing specific primers sequences. B. dendrobatidis was detected in 61/107 (57% and 18/107 (17% animals, respectively by nested and singleplex-PCR. Nested-PCR was statistically more sensible than the conventional for the detection of B. dendrobatidis (Chi-square = 37.1; α = 1% and the agreement between both techniques was considered just fair (Kappa = 0.27. The high prevalence obtained confirms that these fungi occur in free-living frogs from the Brazilian Atlantic Forest with no macroscopic lesions, characterizing the state of asymptomatic carrier. We concluded that the nested-PCR technique, due to its ease of execution and reproducibility, can be recommended as one of the alternatives in epidemiological surveys to detect B. dendrobatidis in healthy free-living frog populations.

  11. Writing clear animal activity proposals.

    Science.gov (United States)

    Pinson, David M

    2011-06-01

    Although IACUC-related topics are frequently discussed in the literature, there is little published information about how to write animal activity proposals. In this article, the author discusses key considerations in the writing and review of animal activity proposals. The author then describes a framework for developing and writing clear animal activity proposals that highlight animal welfare concerns. Though these recommendations are aimed at individuals writing and reviewing research proposals, the framework can be modified for other types of animal activity proposals.

  12. Comparison of different protocols for DNA preparation and PCR amplification of mitochondrial genes of tardigrades

    Directory of Open Access Journals (Sweden)

    Ralph O. SCHILL

    2007-09-01

    Full Text Available Phylogenetic relationships and molecular taxonomy within the Tardigrada have been given a lot of attention in recent years. Here I present the first comparison of different protocols for DNA preparation by investigating six commercial available DNA extraction kits and the CTAB method. Successful extraction of DNA from tardigrades depends strongly on the life-stage (embryo, adult, and on the condition of the specimens, respectively on the preservation (anhydrobiotic, ethanol. Although the extraction kits showed differences in the amount of extracted DNA, in all cases fresh tissue of live animals or embryos resulted in the best quality and quantity of DNA. A lesser amount of DNA was extractable from anhydrobiotic animals and embryos and the results of specimens fixed in ethanol were unsatisfactory. All used commercially available DNA extraction kits and PCR cocktails have been focused on vertebrate tissues, blood, cultured cells, bacteria and yeast. However, I used successfully the kits according to the manufacturer’s instruction without changes in the protocols for DNA extraction of tardigrades. Commercial kits provide a simple and convenient way to isolate pure genomic DNA of high-quality from tardigrades. Furthermore I tested eight different Taq polymerase enzymes for PCR amplification of mitochondrial genes of tardigrades. Each of the enzymes resulted in a PCR product, and even if the amount of the PCR products was quite different, it was possible to use it successful for direct sequencing. Summarizing, the successful PCR of the target DNA depends on the purity and quality of the DNA template and for this the species preservation is more critical than the extraction method or the PCR cocktail which can be optimized.

  13. Detection of Brucella abortus DNA in aborted goats and sheep in Egypt by real-time PCR

    OpenAIRE

    Wareth, Gamal; Melzer, Falk; Tomaso, Herbert; Roesler, Uwe; Neubauer, Heinrich

    2015-01-01

    Background Brucellosis is a major zoonoses affects wide range of domesticated as well as wild animals. Despite the eradication program of brucellosis in Egypt, the disease is still endemic among cattle, buffaloes, sheep, goats, and camels. Results In the present study, abortion occurred naturally among 25 animals (10 cows, 5 buffaloes, 9 Egyptian Baladi goats and 1 ewe) shared the same pasture were investigated by real-time polymerase chain reaction (RT-PCR). DNA of Brucella (B.) abortus was ...

  14. ADVANCES IN ANIMAL WELFARE FOR FREE-LIVING ANIMALS.

    Science.gov (United States)

    2016-04-01

    Over several decades, animal welfare has grown into its own free-standing field of scientific study, from its early beginnings in laboratory animal research to eventually include exhibited animals and farm animals. While it has always been present to some degree, consideration of animal welfare for free-ranging animals has lagged behind, developing as a field of study in the last 20 yr or so. Part of that increase was that animal welfare legislation was finally applied to studies being done on free-ranging animals. But it is the appreciation by the biologists and veterinarians working on wild animals, in which the quality of their results is largely controlled by the quality of the animals they use in their studies, which has resulted in increased attention to the well-being or welfare of the animals that they use. Other important influences driving the recognition of wildlife welfare have been changes in the public's expectations of how wild animals are dealt with, a shift in focus of wildlife professionals from managing animals that can be hunted or angled to include nongame species, the decrease in participation in hunting and fishing by members of the public, and the entry of large numbers of women into fish and wildlife agencies and departments and into veterinary medicine. Technical improvements have allowed the safe capture and handling of large or dangerous animals as immobilization drugs and equipment have been developed. The increasing use of sedating drugs allows for handling of animals with reduced stress and other impacts. A number of topics, such as toe-clipping, branding, defining which taxa can or cannot feel pain, catch-and-release fishing, and more, remain controversial within wildlife science. How we treat the wild animals that we deal with defines who we are as wildlife professionals, and animal welfare concerns and techniques for free-ranging animals will continue to develop and evolve.

  15. Characterization of some Brucella species from Zimbabwe by biochemical profiling and AMOS-PCR

    Directory of Open Access Journals (Sweden)

    Skjerve Eystein

    2009-12-01

    Full Text Available Abstract Background Bovine brucellosis caused by Brucella abortus is endemic in most large commercial and smallholder cattle farms of Zimbabwe, while brucellosis in other domestic animals is rare. The diagnosis of brucellosis is mainly accomplished using serological tests. However, some Brucella spp. have been isolated from clinical cases in the field and kept in culture collection but their biochemical profiles were not documented. We report biochemical profiling and AMOS-PCR characterization of some of these field isolates of Brucella originating from both commercial and smallholder cattle farming sectors of Zimbabwe. Findings Fourteen isolates of Brucella from culture collection were typed using biochemical profiles, agglutination by monospecific antisera, susceptibility to Brucella-specific bacteriophages and by AMOS-PCR that amplifies species- specific IS711. The results of the biochemical profiles for B. abortus biovar 1 (11 isolates and biovar 2 (2 isolates were consistent with those of reference strains. A single isolate from a goat originating from a smallholder mixed animal farm was identified as B. melitensis biovar 1. The AMOS-PCR produced DNA products of sizes 498 bp and 731 bp for B. abortus (biovar 1 and 2 and B. melitensis biovar 1, respectively. Conclusion We concluded that the biochemical profiles and AMOS-PCR characterization were consistent with their respective species and biovars. B. abortus biovar 1 is likely to be the predominant cause of brucellosis in both commercial and smallholder cattle farms in Zimbabwe.

  16. Determination of pig sex in meat and meat products using multiplex real time-PCR.

    Science.gov (United States)

    Abdulmawjood, A; Krischek, C; Wicke, M; Klein, G

    2012-07-01

    For specific production lines, European retail companies demand exclusively female pork meat. To control the quality of their suppliers the identification and a quantitative detection of the animal sex origin of the meat is therefore of importance for meat processors. To enable a fast and reliable detection of male pig meat, a real time-PCR-system was designed in the present study. This was based on the genes AMEL-X and AMEL-Y. The real time-PCR assay allowed the detection of male pig meat at a concentration of 1% yielding a detection probability of 100% while the detection probability investigating meat samples containing 0.1% male pig meat was 44.4%. The analytic sensitivity of this system was assessed to be PCR reaction. The assessment of the accuracy of the real time-PCR assay to correctly identify sex individuals was investigated with 62 pigs including males (n=29) and females (n=33) belonging to different breeds/lines. With the newly designed test all analysed animals were correctly sexed. No amplification was obtained with cow, goat, sheep, turkey and chicken genomic DNA. The presented assay can be used for sex diagnosis, for the detection of male pig meat and for meat quality control.

  17. Prevalence of Arcobacter spp. in humans, animals and foods of animal origin including sea food from India.

    Science.gov (United States)

    Patyal, A; Rathore, R S; Mohan, H V; Dhama, K; Kumar, A

    2011-10-01

    The present study reports the prevalence of Arcobacter, an emerging pathogen in human, animals and foods of animal origin in India. A total of 600 samples from various sources, viz. diarrhoeal stools of humans and dogs, faecal swabs of animals (pig, poultry), preputial washings of breeding bulls and food samples (chicken, pork, fish) were examined for presence of Arcobacter spp. Using cultural methods, a total of 63 Arcobacter spp. were isolated of 600 (10.50%) samples with highest isolation rate were from pig faeces (21.33%) followed by sea foods (17.33%), poultry faeces (14.67%), pork (16.00%), chicken meat (12.00%) and human stools (2.67%). The isolates were confirmed as arcobacters by genus-based PCR. PCR screening of all the enriched samples revealed the overall prevalence of Arcobacter spp. to be 12.00% with highest in pig (25.33%), followed by sea food (21.33%), poultry (17.33%), pork (16%), chicken meat (12%) and human stools (4.00%). No Arcobacter spp. was isolated or detected from diarrhoeal faecal samples of dogs and preputial washings. With multiplex PCR, three different species were detected (A. butzleri, A. cryaerophilus and A. skirrowii) with most of the samples showing mixed infections. There are only two recent reports from India; one with cultural isolation and another with PCR detection of Arcobacter spp. in stool samples of humans with clinical diarrhoea. In this context, our present report is the first report of isolation and detection of Arcobacter spp. from various sources of animals and foods including diarrhoeic human stool samples, utilizing both cultural and molecular tools identifying arcobacters at genus and species level. These results support the importance of arcobacters as an emerging food-borne pathogen, possessing zoonotic potential.

  18. Production of positive controls for calcivirus-specific PCR using recombinant baculiovirus technology

    OpenAIRE

    Butcher, S.A.; Gould, E. A.

    1997-01-01

    Recent advances in our knowledge of the genetic structure of human caliciviruses (HuCVs) and small round-structured viruses (SRSVs) have led to the development of polymerase chain reaction (PCR)-based molecular tests specific for these viruses. These methods have been developed to detect a number of human pathogenic viruses in environmental samples including water, sewage and shellfish. HuCVs and SRSVs are not culturable, and no animal model is currently available. Therefore there is no conve...

  19. Browsed twig environmental DNA: diagnostic PCR to identify ungulate species.

    Science.gov (United States)

    Nichols, Ruth V; Königsson, Helena; Danell, Kjell; Spong, Göran

    2012-11-01

    Ungulate browsing can have a strong effect on ecological processes by affecting plant community structure and composition, with cascading effects on nutrient cycling and animal communities. However, in the absence of direct observations of foraging, species-specific foraging behaviours are difficult to quantify. We therefore know relatively little about foraging competition and species-specific browsing patterns in systems with several browsers. However, during browsing, a small amount of saliva containing buccal cells is deposited at the bite site, providing a source of environmental DNA (eDNA) that can be used for species identification. Here, we describe extraction and PCR protocols for a browser species diagnostic kit. Species-specific primers for mitochondrial DNA were optimized and validated using twigs browsed by captive animals. A time series showed that about 50% of the samples will amplify up to 12 weeks after the browsing event and that some samples amplify up to 24 weeks after browsing (12.5%). Applied to samples of natural browsing from an area where moose (Alces alces), roe deer (Capreolus capreolus), fallow deer (Cervus dama) and red deer (Cervus elaphus) are sympatric, amplification success reached 75%. This method promises to greatly improve our understanding of multispecies browsing systems without the need for direct observations.

  20. Resolución de equivalencias financieras mediante ecuaciones con coeficientes borrosos

    Directory of Open Access Journals (Sweden)

    Maria Silvia Moriñigo

    2007-01-01

    Full Text Available A menudo sólo se conocen estimaciones de las variables financieras. Es usual que con objeto de utilizar modelos clásicos, apreciaciones como “una tasa de entre el 5% y el 7%”, se conviertan en cantidades exactas, como puede ser el promedio entre los valores extremos. En este trabajo se propone un enfoque más flexible que permite captar la incertidumbre mediante la utilización de algunos elementos de la teoría de los conjuntos borrosos. La imprecisión presente en el capital, interés y/o cantidad de períodos se modela mediante números borrosos triangulares. Al apelar a los enfoques clásicos para evaluar expresiones algebraicas con coeficientes borrosos que hacen uso del principio de extensión y aritmética de -cortes, se definen las extensiones fuzzy de las relaciones financieras elementales. Se obtienen las versiones fuzzy del valor actual y del valor final de un capital borroso y el VAN mediante la resolución de ecuaciones con coeficientes borrosos por el método de α - cortes. Estos desarrollos se aplican a distintos casos de estudio. Por último, se muestra que no siempre es posible hallar un análogo borroso de la TIR utilizando los métodos clásicos de resolución de ecuaciones con coeficientes borrosos. Se calcula valiéndose de un nuevo concepto de solución

  1. Biotechnology and DNA vaccines for aquatic animals

    Science.gov (United States)

    Kurath, G.

    2008-01-01

    Biotechnology has been used extensively in the development of vaccines for aquaculture. Modern molecular methods such as polymerase chain reaction (PCR), cloning and microarray analysis have facilitated antigen discovery, construction of novel candidate vaccines, and assessments of vaccine efficacy, mode of action, and host response. This review focuses on DNA vaccines for finfish to illustrate biotechnology applications in this field. Although DNA vaccines for fish rhabdoviruses continue to show the highest efficacy, DNA vaccines for several other viral and bacterial fish pathogens have now been proven to provide significant protection against pathogen challenge. Studies of the fish rhabdovirus DNA vaccines have elucidated factors that affect DNA vaccine efficacy as well as the nature of the fish innate and adaptive immune responses to DNA vaccines. As tools for managing aquatic animal disease emergencies, DNA vaccines have advantages in speed, flexibility, and safety, and one fish DNA vaccine has been licensed.

  2. Real-time PCR: Advanced technologies and applications

    Science.gov (United States)

    This book brings together contributions from 20 experts in the field of PCR, providing a broad perspective of the applications of quantitative real-time PCR (qPCR). The editors state in the preface that the aim is to provide detailed insight into underlying principles and methods of qPCR to provide ...

  3. Detection of Cryptosporidium and Giardia in agricultural and water environments in the Qinghai area of China by IFT and PCR.

    Science.gov (United States)

    Ma, Liqing; Sotiriadou, Isaia; Cai, Qigang; Karanis, Gabriele; Wang, Geping; Wang, Guanghua; Lu, Yan; Li, Xiuping; Karanis, Panagiotis

    2014-09-01

    Qinghai Province in northwest China is strongly influenced by agricultural activities and is an important source of food and drinking water. Here, we present findings regarding the occurrence and molecular epidemiology of Cryptosporidium and Giardia species based on a large-scale investigation of areas of Qinghai Province. The diagnosis and molecular detection of Cryptosporidium oocysts and Giardia cysts was carried out using immunofluorescence microscopy (IFT), whereas nested polymerase chain reaction (PCR) in fecal smears and water samples was used for the detection and molecular characterization of the species. In total, 561 samples (260 water samples and 301 fecal samples from animals) were collected and analyzed. Of the 260 water samples, 66 samples were Cryptosporidium-positive by IFT and 71 samples were positive by nested PCR; in addition, 39 samples were Giardia-positive by IFT and 40 samples were positive by nested PCR. Of the 301 fecal samples from animals, 98 samples were Cryptosporidium-positive by IFT and 61 samples were positive by nested PCR, whereas 52 samples were Giardia-positive by IFT and 31 samples were positive by nested PCR. We showed that the water supplies and animals investigated contained Cryptosporidium and Giardia (oo)cysts. Thus, we recommend that the Chinese Government and Chinese health authorities undertake control measures to protect the food and drinking water sources in Qinghai from these pathogenic protozoa.

  4. Clasificación automática mediante la CDU con el procedimiento en cadena

    OpenAIRE

    San Segundo Manuel, Rosa

    2002-01-01

    Actas de las I Jornadas de Tratamiento y Recuperación de Información (JOTRI), Valencia, España, 4-5 julio 2002 Se entiende por clasificación automática el proceso de agrupar según el contenido las referencias de los documentos o bien los propios documentos electróneos. Este proceso se realiza mediante programas capaces de comparar términos empleados utilizados en el documento. E incluso hay otras formas automáticas de clasificación que emplean procedimientos auto...

  5. Materiales de carbono para desalinización de aguas mediante desionización capacitiva

    OpenAIRE

    Díaz Baizán, Patricia

    2012-01-01

    El presente trabajo muestra los resultados obtenidos al emplear materiales de carbono de elevada área superficial en el proceso de desalinización de aguas mediante desionización capacitiva (CDI). Esta técnica electroquímica, basada en la formación de la doble capa eléctrica en la interfase electrodo/electrolito para retener las sales presentes en el agua tratada, es una alternativa energéticamente menos costosa a las tecnologías comúnmente empleadas como son la ósmosis inversa y la destilació...

  6. Terapia de exposición mediante Realidad Virtual en los trastornos alimentarios: presente y futuro

    OpenAIRE

    Ferran Vilalta Abella; Jana Pla Sanjuanelo; Marta Ferrer García; José Gutiérrez Maldonado

    2015-01-01

    El objetivo de este trabajo fue revisar el uso de la realidad virtual (RV) como técnica de exposición para el tratamiento de los trastornos de la conducta alimentaria (TCA). Para ello se analizó la literatura existente con el fin de responder a dos cuestiones: primero, ¿existe evidencia sobre la capacidad de los entornos y estímulos virtuales para provocar respuestas de ansiedad y craving en pacientes con TCA?; y, segundo, ¿la aplicación de técnicas de exposición mediante RV permite la dismin...

  7. Conocer el valor de la expresión corporal mediante la danza

    OpenAIRE

    Ochoa Ullate, Garbiñe

    2014-01-01

    Mediante este trabajo se pretende trabajar una serie de actividades y ejercicios para conocer la expresión corporal y la salud a través de de diferentes danzas africanas. Para la realización del trabajo se analizan las principales aportaciones pedagógicas de varios autores relacionados con la enseñanza de estos conceptos. Se trata de un trabajo empírico, con el cual se pretende analizar y estudiar los resultados de la propuesta didáctica para llegar a unas conclusiones acerca de si el ámbi...

  8. Tratamiento del tortícolis congénito mediante control estimular y fisioterapia

    OpenAIRE

    MARÍA ASUNCIÓN VÁZQUEZ VILÀ; SUSANA COLLADO VÁZQUEZ

    2004-01-01

    Presentamos un caso clínico de tortícolis congénito y su tratamiento mediante control estimular y fisioterapia, por la alta frecuencia del tortícolis en la práctica clínica y por los buenos resultados que en general presenta el tratamiento conservador en estos casos, principalmente cuando éste se inicia de forma precoz. El caso presentado es de una niña con tortícolis congénito izquierdo, sin patologías asociadas, en tratamiento desde los 15 días hasta los nueve meses de edad, con una buena e...

  9. Ensayo no destructivo de soldaduras en pernos conectores mediante inspección acústica

    Directory of Open Access Journals (Sweden)

    Aznar, A.

    2012-09-01

    Full Text Available Headed studs are nowadays the standard steel-concrete connectors because of their competitive advantages. Firstly, they provide a high degree of safety thanks to semiautomatic electric arc welding. These welds are not suitable for typical non-destructive tests. The analytical study comprises several models. The first vibration modes have been obtained. The experimental research has developed first the measurement of the natural frequencies of 28 headed-studs in the sonic range. Then they have been tested by non-destructive and destructive tests. Finally theirs tests have been compared with their respective frequency measurements. A clear relationship between the measured frequencies and the lack of penetration of the welds has been established, that confirms the analytical prediction of this effect of the internal weld imperfections. Therefore, the feasibility of simple and absolutely non-destructive tests of welded studs by in site measurement of natural frequencies in the sonic range has been clearly established in this work.

    Los pernos conectores aportan múltiples ventajas de uso, entre las que se encuentra el elevado margen de seguridad que ofrecen sus soldaduras ejecutadas mediante arco eléctrico. Estas soldaduras, aunque ampliamente fiables, son difícilmente comprobadas mediante ensayos no destructivos. El presente estudio plantea la inspección de soldaduras de pernos conectores mediante su espectro acústico. Analíticamente, la investigación se ha centrado en el cálculo de los primeros modos propios de vibración. Experimentalmente se han medido las frecuencias propias de resonancia de 28 pernos, en los que posteriormente se han llevado a cabo ensayos tanto no destructivos como destructivos. Se ha obtenido, tanto teórica como experimentalmente, una relación entre la frecuencia de vibración de los pernos conectores y la calidad de la soldadura. Por ello se verifica la posibilidad de inspección de estas

  10. PREDICCION DE PROPIEDADES DE MEZCLAS GASOLINA-ETANOL MEDIANTE ESPECTROSCOPIA DE INFRARROJO

    OpenAIRE

    Felipe Sánchez-Minero; Edgar Ramirez-Jimenez; Guadalupe Silva-Oliver; Berenice Hernandez-Castillejos

    2013-01-01

    El efecto del etanol en la reformulación de una gasolina fue estudiado mediante espectroscopia de infrarrojo (IR). Para ello, se prepararon seis mezclas gasolina-etanol con diferente contenido de alcohol (0, 20, 40, 60, 80 y 100% volumen). Los espectros IR de estas mezclas fueron obtenidos en la región de 4000 a 850 cm -1. A continuación, se determinó un factor característico del etanol (F CO), el cual relaciona el área bajo la curva de la banda a 1050 cm-1 y el área bajo la curva total del e...

  11. Diseño de una banda transportadora mediante guide de Matlab

    OpenAIRE

    Salinero Gervaso, Mario

    2013-01-01

    El objetivo principal del presente proyecto, es la elaboración de un programa para diseñar una banda transportadora mediante GUIDE de MatLab, conforme a las normas: DIN 22101, ISO 5048 y UNE 58204. Otro objetivo (secundario), por el que se ha desarrollado este software, es ayudar a los estudiantes de Ingeniería Industrial en el cálculo y diseño de una cinta transportador, para así facilitarles el entendimiento de parte del temario de la asignatura de "Transportes en la Ingen...

  12. Caracterización de bacterias aisladas en larvas de p.v. mediante rapds

    OpenAIRE

    Cedeño, Ricardo

    2003-01-01

    Caracterización de bacterias aisladas en larvas de P.V. mediante RAPDs La “Vibriosis luminiscente” reportada por varios laboratorios de larvas del Ecuador es asociada principalmente a la bacteria Vibrio harveyi. Los métodos de identificación bacteriana tradicionales se basan en la observación de características morfológicas y pruebas bioquímicas, sin embargo la identificación de especies bacterianas y en especial de Vibrios ambientales ha presentado ciertas dificultades debido al tiempo qu...

  13. PROMOCIÓN DE LA LECTURA E IDENTIDAD DEPORTIVA MEDIANTE TEXTOS DE HISTORIA DEL DEPORTE

    OpenAIRE

    2016-01-01

    En el artículo se expone una experiencia de promoción de la lectura desarrollada mediante un proyecto conjunto de la Facultad de Cultura Física y la biblioteca de la Universidad de Las Tunas. Se fundamenta en aspectos teóricos de la promoción de la lectura y de la identidad deportiva, ambas variables tienen salida práctica en actividades donde confluyen diversas manifestaciones del arte y la literatura con la actividad física y el deporte. El objetivo del trabajo consiste en reflexionar en to...

  14. Aprendizaje de procesos analíticos mediante clases prácticas con laboratorios virtuales

    OpenAIRE

    Sierra Fernández, J.; J. Bosque Sendra; García Campaña, A.; Blanc García, R.; Del Iruela, M.; Gámiz Gracia, L.; Alés Barrero, F.

    2009-01-01

    Este trabajo forma parte de un proyecto de innovación docente financiado por la Universidad de Granada durante el curso 2008-09 y pretende que el alumnado del Departamento de Química Analítica de los primeros cursos de Química, Ingeniería Química, Ciencias Ambientales y Farmacia sea capaz de interrelacionar los aspectos teóricos y prácticos de diversos procesos analíticos mediante el uso de laboratorios virtuales en las sesiones prácticas. Con este fin, las clases prácticas son planificadas y...

  15. Superación de la dislexia mediante técnicas operantes

    OpenAIRE

    Bernardo Gutiérrez, Inmaculada; Pérez Álvarez, Marino

    1993-01-01

    Numerosos trabajos e investigaciones han tratado de establecer una relación casi causal entre los déficits psicomotrices – lateralidad, orientación, ritmo, etc. – y la dislexia. El presente estudio se dirige a analizar mediante una intervención experimental esta supuesta relación, sobre ta que se sustenta, por otra parte, el tratamiento psicoeducativo tradicional. Se ha aplicado, así, una intervención psicomotriz (paquete terapéutico estándar) y otra conductual (específicamente lectora) a una...

  16. Regeneración ósea mediante estímulos mecánicos

    OpenAIRE

    Cocera Castillo, Daniel

    2013-01-01

    RESUMEN: Recientes investigaciones sugieren que la estimulación mecánica del tejido óseo puede provocar la regeneración y el refuerzo del mismo. A partir de esta hipótesis, se han analizado las distintas posibilidades de tratamiento o entrenamiento del hueso mediante técnicas manuales. Entre los resultados más destacables se encuentran como las cargas mecánicas son las responsables de una mejor salud del hueso, siendo capaces de ayudar en la regeneración de fracturas y aumentar la densidad en...

  17. DESULFURACIÓN DE RELAVE MEDIANTE LA FLOTACIÓN DE SULFUROS DE HIERRO

    OpenAIRE

    Luis Valderrama Campusano; B. Zazzali; J. Chamorro; Santander, M.

    2015-01-01

    En Chile la minería se desarrolla principalmente en  la concentración de cobre, molibdeno, oro y plata, mediante el proceso de flotación. Este genera grandes tonelajes de relaves que contienen diversas minerales, principalmente pirita. Dado que estos minerales se depositan en los tranques, estos pueden generar aguas ácidas; se propone como alternativa la desulfuración de los súlfuros por flotación. En una primera parte se estudió la flotabilidad de la pirita en un tubo Hallimond, utilizando c...

  18. Foundation Flash Cartoon Animation

    CERN Document Server

    Jones, Tim; Rosson, Allan S

    2008-01-01

    One of Flash s most common uses is still animation for cartoons, games, advertising etc, and this book takes a fresh look at the topic, breaking it down pre-production, production, and post production, and looking at each section in detail, and covering topics such as storyboarding, character libraries and camera mechanics like no Flash book has before. The book is written by members of the Emmy award winning ANIMAX team, who have created work for clients such as Disney, AOL, Fox, WWE, ESPN, and Sesame workshop. This book is an opportunity for them to share their secrets, and is written to sui

  19. Animation-based Sketching

    DEFF Research Database (Denmark)

    Vistisen, Peter

    This thesis is based on the results of a three-year long PhD-study at the Department of Communication and Psychology at Aalborg University. The thesis consist of five original papers, a book manuscript, as well as a linking text with the thesis’ research questions, research design, and summary...... experiments has been carried out, applying animation-based sketching in various contexts and at varying points in the design process. In the studies, I evaluate the viability of the approach, the practical integration into the design process, and map how consensus between stakeholders in design can...

  20. Storyboarding an Animated Film

    DEFF Research Database (Denmark)

    Frølunde, Lisbeth

    2009-01-01

    This paper applies notions of transformation to the analysis of data on semiotic processes related to making an animated film. The data derives from a study conducted in an upper secondary school in Copenhagen with students (18 years old) participating in a week-long workshop. The paper applies...... the concept of transduction with a focus on film storyboards: how students transform ideas when working with different modes (audio, visual) of representation. Data includes discourse analysis of semiotic processes and texts, referring to Social Semiotics and the methodology of Mediated Discourse Analysis...

  1. Telltale Animation (Sol 8)

    Science.gov (United States)

    2008-01-01

    This animation of the NASA's Phoenix Mars Lander's telltale was made from five images taken by Phoenix's Stereo Surface Imager (SSI) just after 1:10 PM local Mars time on the eighth Martian day of the mission, or Sol 8 (June 2, 2008). The images were taken with a blue filter (450 nanometer, R6) that focuses at items on the deck rather than the workspace or horizon. The Phoenix Mission is led by the University of Arizona, Tucson, on behalf of NASA. Project management of the mission is by NASA's Jet Propulsion Laboratory, Pasadena, Calif. Spacecraft development is by Lockheed Martin Space Systems, Denver.

  2. Telltale Animation (Sol 9)

    Science.gov (United States)

    2008-01-01

    This animation of the NASA's Phoenix Mars Lander's telltale was made from five images taken by Phoenix's Stereo Surface Imager (SSI) just after 4:37 PM local Mars time on the ninth Martian day of the mission, or Sol 9 (June 3, 2008). The images were taken with a blue filter (450 nanometer, R6) that focuses at items on the deck rather than the workspace or horizon. The Phoenix Mission is led by the University of Arizona, Tucson, on behalf of NASA. Project management of the mission is by NASA's Jet Propulsion Laboratory, Pasadena, Calif. Spacecraft development is by Lockheed Martin Space Systems, Denver.

  3. Aspergillus PCR: one step closer to standardization.

    NARCIS (Netherlands)

    White, P.L.; Bretagne, S.; Klingspor, L.; Melchers, W.J.G.; McCulloch, E.; Schulz, B.; Finnstrom, N.; Mengoli, C.; Barnes, R.A.; Donnelly, J.P.; Loeffler, J.

    2010-01-01

    PCR has been used as an aid in the diagnosis of invasive aspergillosis for almost 2 decades. A lack of standardization has limited both its acceptance as a diagnostic tool and multicenter clinical evaluations, preventing its inclusion in disease-defining criteria. In 2006, the European Aspergillus P

  4. The Power of Real-Time PCR

    Science.gov (United States)

    Valasek, Mark A.; Repa, Joyce J.

    2005-01-01

    In recent years, real-time polymerase chain reaction (PCR) has emerged as a robust and widely used methodology for biological investigation because it can detect and quantify very small amounts of specific nucleic acid sequences. As a research tool, a major application of this technology is the rapid and accurate assessment of changes in gene…

  5. PCR clonality detection in Hodgkin lymphoma.

    NARCIS (Netherlands)

    Hebeda, K.M.; Altena, M.C. van; Rombout, P.D.M.; Krieken, J.H.J.M. van; Groenen, P.J.T.A.

    2009-01-01

    B-cell clonality detection in whole tissue is considered indicative of B-cell non-Hodgkin lymphoma (NHL). We tested frozen tissue of 24 classical Hodgkin lymphomas (cHL) with a varying tumor cell load with the multiplex polymerase chain reaction (PCR) primer sets for IGH and IGK gene rearrangement (

  6. Real Time PCR: Principles and Application

    Directory of Open Access Journals (Sweden)

    Safie Amini

    2005-09-01

    Full Text Available The polymerase chain reaction (PCR has been used as the new golden standard for detecting a wide variety of templates across a range of scientific specialties and also as an essential tool in research laboratories. PCR has completely revolutionized the detection of RNA and DNA viruses(1. Real Time vs. Traditional PCRReal time chemistry allows the detection of PCR amplification during the early phase of the reaction. Measuring the kinetic of the reaction in the early phase of PCR provides a distinct advantage over traditional PCR detection. Traditional methods use agarose gel electrophoresis for detection of PCR amplification at the final phase or end point. End point detection is really time consuming; it takes several hours to have the result. On the other hand, results are based on size discrimination. Also, the result of end point is variable from sample to sample. While gels may not resolve this variability in yield, real time PCR is sensitive enough to detect this change.Some problems with end point detection are: poor precision, low sensitivity, short dynamic range (<2 log, low resolution, non-automated procedure, size-based discrimination only, and post PCR processing (carry-over contamination and results are not expressed as numbers(2.Detection of PCR Products in Real-timeReal-time PCR and RT-PCR allow accurate quantification of starting amounts of DNA, cDNA, and RNA targets. Fluorescence is measured during each cycle, which greatly increases the dynamic range of the reaction since the amount offluorescence is proportional to the amount of PCR product. PCR products can be detected using either fluorescent dyes that bind to double-stranded DNA or fluorescently labeled sequence-specific probes(3.SYBR Green ISYBR® Green I binds all double-stranded DNA molecules, emitting a fluorescent signal of a defined wavelength on binding. The excitation and emission maxima of SYBR Green I are at 494 nm and 521 nm, respectively, and are compatible for

  7. Discussing Animal Rights and Animal Research in the Classroom.

    Science.gov (United States)

    Herzog, Harold A.

    1990-01-01

    Reviews two prominent philosophical justifications for animal liberation and describes a simulation that facilitates class discussion of animal research issues. Students reported that the exercise increased their awareness of the issues and of the complexity of making ethical decisions. (DB)

  8. How Many Microorganisms Are Present? Quantitative Reverse Transcription PCR (qRT-PCR)

    Science.gov (United States)

    Price, Andy; Álvarez, Laura Acuña; Whitby, Corinne; Larsen, Jan

    Quantitative reverse transcription PCR (qRT-PCR) is a variation of conventional quantitative or real-time PCR, whereby mRNA is first converted into the complementary DNA (cDNA) by reverse transcription, the cDNA is then subsequently quantified by qPCR. The use of mRNA as the initial template allows the quantification of gene transcripts, rather than gene copy numbers. mRNA is only produced by actively metabolising cells and is produced by its corresponding gene to provide a 'blueprint' in order for a cell to manufacture a specific protein. Conventional qPCR detects not only DNA present in actively metabolising cells but also inactive and dead cells. qRT-PCR has the advantage that only actively metabolising cells are detected, hence provides a more reliable measure of microbial activity in oilfield samples. When qRT-PCR is combined with primers and probes for specific genes, the activity of microbial processes important in the oilfield, such as sulphate reduction, methanogenesis and nitrate reduction can be monitored.

  9. Digital PCR to assess gene-editing frequencies (GEF-dPCR) mediated by designer nucleases.

    Science.gov (United States)

    Mock, Ulrike; Hauber, Ilona; Fehse, Boris

    2016-03-01

    Genome editing using designer nucleases such as transcription activator-like effector nucleases (TALENs) or clustered regularly interspersed short palindromic repeats (CRISPR)-Cas9 nucleases is an emerging technology in basic and applied research. Whereas the application of editing tools, namely CRISPR-Cas9, has recently become very straightforward, quantification of resulting gene knockout rates still remains a bottleneck. This is particularly true if the product of a targeted gene is not easily detectable. To address this problem, we devised a novel gene-editing frequency digital PCR (GEF-dPCR) technique. GEF-dPCR exploits two differently labeled probes that are placed within one amplicon at the gene-editing target site to simultaneously detect wild-type and nonhomologous end-joining (NHEJ)-affected alleles. Taking advantage of the principle of dPCR, this enables concurrent quantification of edited and wild-type alleles in a given sample. We propose that our method is optimal for the monitoring of gene-edited cells in vivo, e.g., in clinical settings. Here we describe preparation, design of primers and probes, and setup and analysis of GEF-dPCR. The setup of GEF-dPCR requires up to 2 weeks (depending on the starting point); once the dPCR has been established, the protocol for sample analysis takes <1 d.

  10. Clinical Identification of Common Species of Dermatophytes by PCR and PCR-RFLP

    Institute of Scientific and Technical Information of China (English)

    丁娟; 李家文; 刘志香; 谭志建

    2004-01-01

    To find a fast and efficient way of identifying seven common dermatophytes in clinical practice, we used the techniques of polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (RFLP) targeting Topoisomerase Ⅱ gene. The DNA of 7 dermatophytes, along with Candida albicans, Aspergillus terreus and Aspergillus flavus were amplified by consensus primer dPsD1. They were then subjected to a second PCR with primers dPsD2 and species-specific primers PsT and PsME separately. 6 of the products generated by dPsD2 were digested with restriction enzyme Hinc Ⅱ. DNA fragments of 3390 bp and 2380 bp was amplified by using consensus primer dPsD1 and dPsD2 from the genomic DNA of each dermatophyte species separately. By combining the results of the two species-specific primer sets (PsT and PsME), all species of dermatophyte yielded unique sizes-set of PCR products expect for T. mentagrophytes and T. tonsurans.From the restriction profiles of Hinc Ⅱ , 6 of the 7 dermatophytoses were diagnosed to species level including T. mentagrophytes and T. tonsurans. By combining the results of the PCR and PCRRFLP, the 7 common dermatophytes can be identified to species level. It is conclude that the multiplex PCR and PCR-RFLP identification targeting the DNA topoisomerase Ⅱ gene is rapid and efficient.

  11. Quantitative PCR and Digital PCR for Detection of Ascaris lumbricoides Eggs in Reclaimed Water

    Directory of Open Access Journals (Sweden)

    Lucrecia Acosta Soto

    2017-01-01

    Full Text Available The reuse of reclaimed water from wastewater depuration is a widespread and necessary practice in many areas around the world and must be accompanied by adequate and continuous quality control. Ascaris lumbricoides is one of the soil-transmitted helminths (STH with risk for humans due to its high infectivity and an important determinant of transmission is the inadequacy of water supplies and sanitation. The World Health Organization (WHO recommends a limit equal to or lower than one parasitic helminth egg per liter, to reuse reclaimed water for unrestricted irrigation. We present two new protocols of DNA extraction from large volumes of reclaimed water. Quantitative PCR (qPCR and digital PCR (dPCR were able to detect low amounts of A. lumbricoides eggs. By using the first extraction protocol, which processes 500 mL of reclaimed water, qPCR can detect DNA concentrations as low as one A. lumbricoides egg equivalent, while dPCR can detect DNA concentrations as low as five A. lumbricoides egg equivalents. By using the second protocol, which processes 10 L of reclaimed water, qPCR was able to detect DNA concentrations equivalent to 20 A. lumbricoides eggs. This fact indicated the importance of developing new methodologies to detect helminth eggs with higher sensitivity and precision avoiding possible human infection risks.

  12. Ethical Inspection about laboratory animals.

    Science.gov (United States)

    Yang, Nai-bin; Pan, Xiao-jun; Cheng, Jing-jing; Lin, Jia-qiang; Zhu, Jia-yin

    2015-11-01

    Laboratory animals and animal experiments are foundations and important support conditions for life sciences, especially for medical research. The animal experiments have drawn extensive attention from the society because of the ethical issue. This paper takes Wenzhou Medical University as an example to give a brief introduction to the ethical review about laboratory animals in the university so as to further draw attention and concerns from the public about the ethical issue of laboratory animals. We successively introduce its scientific projects, nurturing environment and ethical review of laboratory animals.

  13. Detection of Brucella species in the milk of infected cattle, sheep, goats and camels by PCR.

    Science.gov (United States)

    Hamdy, Mahmoud E R; Amin, A S

    2002-05-01

    One hundred and three milk samples were collected from 52 cows, 21 ewes, 18 goats and 12 camels. The animals tested positive to at least one of the following: (1) standard tube agglutination test (SAT); (2) Rose Bengal plate test (RBPT); (3) milk ring test (MRT). All milk samples were examined by culture and single-step polymerase chain reaction (PCR) techniques for detection of Brucella species. The PCR assay amplified Brucella-DNA from 29 bovine milk samples, 10 from sheep, 13 from goats and one from a camel. The direct culture method detected Brucella organisms from 24 samples of cows' milk, 12 from sheep, 10 from goats and failed to detect any Brucella organisms from camels' milk. PCR detected up to 100 colony forming units (CFU) of B. abortus per millilitre of milk in 100% of diluted milk samples, and 1000 CFU of B. melitensis from 70% of milk samples. Although the overall sensitivity of the PCR was higher than the culture method, it should be possible to increase the sensitivity to detect lower numbers of Brucella organisms in field samples. The speed and sensitivity of the PCR assay suggest that this technique could be useful for detection of Brucella organisms in bovine milk, as well as in sheep, goat, and camels milk.

  14. Murine model of disseminated fusariosis: evaluation of the fungal burden by traditional CFU and quantitative PCR.

    Science.gov (United States)

    González, Gloria M; Márquez, Jazmín; Treviño-Rangel, Rogelio de J; Palma-Nicolás, José P; Garza-González, Elvira; Ceceñas, Luis A; Gerardo González, J

    2013-10-01

    Systemic disease is the most severe clinical form of fusariosis, and the treatment involves a challenge due to the refractory response to antifungals. Treatment for murine Fusarium solani infection has been described in models that employ CFU quantitation in organs as a parameter of therapeutic efficacy. However, CFU counts do not precisely reproduce the amount of cells for filamentous fungi such as F. solani. In this study, we developed a murine model of disseminated fusariosis and compared the fungal burden with two methods: CFU and quantitative PCR. ICR and BALB/c mice received an intravenous injection of 1 × 10(7) conidia of F. solani per mouse. On days 2, 5, 7, and 9, mice from each mice strain were killed. The spleen and kidneys of each animal were removed and evaluated by qPCR and CFU determinations. Results from CFU assay indicated that the spleen and kidneys had almost the same fungal burden in both BALB/c and ICR mice during the days of the evaluation. In the qPCR assay, the spleen and kidney of each mouse strain had increased fungal burden in each determination throughout the entire experiment. The fungal load determined by the qPCR assay was significantly greater than that determined from CFU measurements of tissue. qPCR could be considered as a tool for quantitative evaluation of fungal burden in experimental disseminated F. solani infection.

  15. ERIC-PCR genotypic characterization of Haemophilus parasuis isolated from Brazilian swine.

    Science.gov (United States)

    Moreno, Luisa Zanolli; Castilla, Karina Salvagni; de Gobbi, Débora Dirani Sena; Coutinho, Tania Alen; Ferreira, Thais Sebastiana Porfida; Moreno, Andrea Micke

    2011-10-01

    Haemophilus parasuis infection, known as Glässer's disease, is characterized by fibrinous polyserositis, arthritis and meningitis in piglets. Although traditional diagnosis is based on herd history, clinical signs, bacterial isolation and serotyping, the molecular-based methods are alternatives for species-specific tests and epidemiologic study. The aim of this study was to characterize H. parasuis strains isolated from different states of Brazil by serotyping, PCR and ERIC-PCR. Serotyping revealed serovar 4 as the most prevalent (24 %), followed by serovars 14 (14 %), 5 (12 %), 13 (8 %) and 2 (2 %), whereas 40 % of the strains were considered as non-typeable. From 50 strains tested 43 (86%) were positive to Group 1 vtaA gene that have been related to virulent strains of H.parasuis. ERIC-PCR was able to type isolates tested among 23 different patterns, including non-typeable strains. ERIC-PCR patterns were very heterogeneous and presented high similarity between strains of the same animal or farm origin. The results indicated ERIC-PCR as a valuable tool for typing H. parasuis isolates collected in Brazil.

  16. [Dermatophytes from animals].

    Science.gov (United States)

    Hasegawa, A

    2000-01-01

    Dermatophytes from animal dermatophytoses were investigated, especially for their teleomorphs and molecular characteristics. Microsporum canis, M. equinum, M. gypseum, M. nanum, Trichophyton equinum, T. mentagrophytes complex, T. rubrum and T. verrucosum were isolated as the etiological agents of animal ringworm in Japan. They were morphologically and biochemically identified and their perfect states were examined. The teleomorph of M. canis was first obtained and reported as Nanizzia otae in 1975. The isolates of M. canis of which the teleomorph was confirmed were all "-" excepted two Japanese isolates. Mating experiments indicated that the isolates of M. gypseum were "+" or "-" of A. gypseum and A. incurvatum, respectively. Most of the isolates of T. mentagrophytes complex of which a perfect state was detected were A. vanbreuseghemii. One isolate from a monkey was A. simii and one from a rabbit was A. benhamiae. However, the teleomorph remained unknown in many isolates. Molecular characteristics in random amplification of polymorphic DNA and Southern hybridization analyses were found to be effective to differentiate the species of Microsporum. Nucleotide sequences of chitin synthase 1 (CHS1) gene of dermatophytes were also analyzed for their phylogenetic relatedness. The phylogenetic analysis revealed four clusters: the first cluster consisted of A. benhamiae, A. simii, A. vanbreuseghemii, T. mentagrophytes var. interdigitale, T. rubrum and T. violaceum; the second of A. fulvum, A. gypseum and A. incurvatum; the third of A. grubyi and A. otae; and the fourth of Epidermaphyton floccosum, providing useful information for the classification and understanding of their evolution.

  17. Replicating animal mitochondrial DNA

    Directory of Open Access Journals (Sweden)

    Emily A. McKinney

    2013-01-01

    Full Text Available The field of mitochondrial DNA (mtDNA replication has been experiencing incredible progress in recent years, and yet little is certain about the mechanism(s used by animal cells to replicate this plasmid-like genome. The long-standing strand-displacement model of mammalian mtDNA replication (for which single-stranded DNA intermediates are a hallmark has been intensively challenged by a new set of data, which suggests that replication proceeds via coupled leading-and lagging-strand synthesis (resembling bacterial genome replication and/or via long stretches of RNA intermediates laid on the mtDNA lagging-strand (the so called RITOLS. The set of proteins required for mtDNA replication is small and includes the catalytic and accessory subunits of DNA polymerase y, the mtDNA helicase Twinkle, the mitochondrial single-stranded DNA-binding protein, and the mitochondrial RNA polymerase (which most likely functions as the mtDNA primase. Mutations in the genes coding for the first three proteins are associated with human diseases and premature aging, justifying the research interest in the genetic, biochemical and structural properties of the mtDNA replication machinery. Here we summarize these properties and discuss the current models of mtDNA replication in animal cells.

  18. Allele-Specific Quantitative PCR for Accurate, Rapid, and Cost-Effective Genotyping.

    Science.gov (United States)

    Lee, Han B; Schwab, Tanya L; Koleilat, Alaa; Ata, Hirotaka; Daby, Camden L; Cervera, Roberto Lopez; McNulty, Melissa S; Bostwick, Hannah S; Clark, Karl J

    2016-06-01

    Customizable endonucleases such as transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) enable rapid generation of mutant strains at genomic loci of interest in animal models and cell lines. With the accelerated pace of generating mutant alleles, genotyping has become a rate-limiting step to understanding the effects of genetic perturbation. Unless mutated alleles result in distinct morphological phenotypes, mutant strains need to be genotyped using standard methods in molecular biology. Classic restriction fragment length polymorphism (RFLP) or sequencing is labor-intensive and expensive. Although simpler than RFLP, current versions of allele-specific PCR may still require post-polymerase chain reaction (PCR) handling such as sequencing, or they are more expensive if allele-specific fluorescent probes are used. Commercial genotyping solutions can take weeks from assay design to result, and are often more expensive than assembling reactions in-house. Key components of commercial assay systems are often proprietary, which limits further customization. Therefore, we developed a one-step open-source genotyping method based on quantitative PCR. The allele-specific qPCR (ASQ) does not require post-PCR processing and can genotype germline mutants through either threshold cycle (Ct) or end-point fluorescence reading. ASQ utilizes allele-specific primers, a locus-specific reverse primer, universal fluorescent probes and quenchers, and hot start DNA polymerase. Individual laboratories can further optimize this open-source system as we completely disclose the sequences, reagents, and thermal cycling protocol. We have tested the ASQ protocol to genotype alleles in five different genes. ASQ showed a 98-100% concordance in genotype scoring with RFLP or Sanger sequencing outcomes. ASQ is time-saving because a single qPCR without post-PCR handling suffices to score

  19. In silico and in vitro evaluation of PCR-based assays for the detection of Bacillus anthracis chromosomal signature sequences

    NARCIS (Netherlands)

    Agren, J.; Hamidjaja, R.A.; Hansen, T.; Ruuls, R.C.; Thierry, S.; Vigre, H.; Janse, I.; Sundström, A.; Segerman, B.; Koene, M.G.J.; Löfström, Ch.; Rotterdam, van B.; Derzelle, S.

    2013-01-01

    Bacillus anthracis, the causative agent of anthrax, is a zoonotic pathogen that is relatively common throughout the world and may cause life threatening diseases in animals and humans. There are many PCR-based assays in use for the detection of B. anthracis. While most of the developed assays rely o

  20. Molecular characterization of Pasteurella multocida isolates obtained from poultry, ruminant, cats and dogs using RAPD and REP-PCR analysis

    Directory of Open Access Journals (Sweden)

    Hesamaddin Shirzad-Aski

    2016-09-01

    Full Text Available In the present study, Randomly Amplified Polymorphic DNA‌ (RAPD and Repetitive Extragenic Palindromic sequence-based Polymerase Chain Reaction (REP-PCR were used to characterize 131 isolates of Pasteurella multocida, originating from different healthy and diseased animal species obtained from several geographical regions of Iran. The RAPD and REP-PCR generated amplified products in the range of 300 to 3400 bp and 200 to 2850 bp, respectively. Among all of the P. multocida isolates, cluster analysis revealed that 63 clusters and nine untypable isolates and 81 clusters and six untypable isolates were produced with RAPD and REP-PCR methods, respectively. The results indicated that the REP-PCR method showed a slightly higher level of discrimination power in differentiating of P. multocida isolates as compared with RAPD. The results showed that a considerable level of genetic diversity exists among P. multocida isolates even in the isolates with the same animal or geographical origins. There was no host- and region-specific pattern. In addition, the isolates obtained from the healthy and diseased animal did not reveal any correlation genotypic profiles, which could be supported by the hypothesis that P. multocida is a strictly opportunistic pathogen. In conclusion, because of a large amount of genetic heterogeneityin the P. multocida isolates, Pasteurellosis may be caused by different clones in the same herd or animal.

  1. Pronósticos de inflación mediante técnicas bayesianas

    Directory of Open Access Journals (Sweden)

    Juan Diego Chavarría

    2015-11-01

    Full Text Available La efectividad de la política monetaria bajo un esquema de metas de inflación como el propuesto por el Banco Central de Costa Rica se basa en buena medida en el correcto y oportuno pronóstico de la inflación a corto y mediano plazo con el fin de diseñar de mejor forma las acciones de política monetaria. Así, el propósito de este trabajo es desarrollar una herramienta complementaria para elaborar pronósticos de inflación mediante un enfoque bayesiano. Para lo anterior se propone la utilización de la metodología Bayesian Model Averaging y de Weighted Average Least Squares. Los modelos de proyección especificados permitirían ampliar y complementar el análisis que se realiza actualmente con el Modelo Macroeconómico de Proyección Trimestral (MMPT del Banco Central de Costa Rica. Como resultado esta investigación muestra que, para datos de periodicidad mensual y a horizontes de pronóstico de 1 a 12 meses, es posible encontrar proyecciones mediante un proceso bayesiano que poseen una mayor capacidad predictiva en relación con aquellas producidas por un modelo autorregresivo.

  2. Graduación mediante funciones Gompertz-Makeham dinámicas.

    Directory of Open Access Journals (Sweden)

    Montes Suay, Francisco

    2004-01-01

    Full Text Available El objetivo de este trabajo es modelar la influencia del tiempo de calendario y la edad, en las probabilidades de muerte, qx. Para ello, se gradúa los datos de mortalidad de la Comunidad Valenciana correspondientes a los años 1980 a 1999. En primer lugar obtenemos el ajuste de las funciones Gompertz-Makeham con términos correspondientes a las variables edad y tiempo mediante modelos lineales generalizados. La elección del modelo que mejor se ajusta a los datos de cada periodo se lleva a cabo atendiendo a criterios de bondad de ajuste y de complejidad del mismo. Se trata de buscar un equilibrio entre ambos aspectos, para lo cual utilizamos medidas y contrastes implementados en S-plus. Del modelo obtenido para cada sexo hacemos un análisis más detallado mediante los tests no paramétricos habituales y análisis gráfico, para concluir la adecuación de éstos a la experiencia de mortalidad.

  3. Introducción de momentos internos de torsión mediante pretensado

    Directory of Open Access Journals (Sweden)

    Johannson, J.

    1975-02-01

    Full Text Available In this article the possibility is being studied of transferring bending moments to the ground by means of torsional strength of a beam. This avoids the need to subject columns to high torsional stresses. This system, by prestressing, induces internal moments which compensate the torsional moment. This reduces, or eliminates, under certain loading cases, the angular deflection due to torsion couples. Primarily, a study is carried out on this type torsion — with the example of a simple hollow beam, in corbel,En este artículo se estudia la posibilidad de transferir momentos al firme mediante la resistencia torsional de una viga, evitándose así soluciones de pilas o pilares solicitados fuertemente por torsión y creando, mediante pretensado, momentos internos que contrarresten directamente al momento torsor actuante, con lo cual se reduce e incluso se elimina, para ciertos casos de carga, el giro de la sección debido al par torsor. En primer lugar se lleva a cabo un estudio de este tipo de torsión con el ejemplo sencillo de una viga hueca, en voladizo, solicitada en un extremo por un par torsor.

  4. Ahorro energético mediante estrategias de iluminación natural optimizadas

    Directory of Open Access Journals (Sweden)

    Puigdomènech Franquesa, Joan

    1986-04-01

    Full Text Available Electrical charges in buildings and specially in those of commercial use, can be diminished by means of natural lighting strategies. Taking the climate features of our country into consideration, it is necessary to prevent the inconveniences caused by an en erg y excess in summer, so solar Controls are needed. The only practical way to achieve the suitable balance between thermal and light needs, so as to get a monthly or annual energetic balance optimization, is to operate with the computer. A programme with such characteristics is described here. Its application gives important sarings in non renouvable energy savings.Mediante estrategias de iluminación natural es posible disminuir las cargas eléctricas de los edificios y en especial los de uso comercial. Dadas las características climáticas de nuestro país es necesario prever los inconvenientes de un exceso de energía en verano, para lo cual es preciso disponer de controles solares. Encontrar el correcto equilibrio entre las necesidades térmicas y lumínicas en base a la optimización del balance energético mensual o anual es únicamente factible mediante el uso del ordenador. Un programa que responde a estas características es descrito en el presente trabajo, obteniéndose con su aplicación importantes ahorros en el consumo de energías no renovables.

  5. PROPUESTA DE CONEXIÓN DE ENTORNOS IPv6 MEDIANTE UN BACKBONE MPLS/IPv4

    Directory of Open Access Journals (Sweden)

    Nancy Yaneth Gelvez García

    2013-09-01

    Full Text Available Las redes actuales MPLS/IPv4 presentan las ventajas de poder implementar ingeniería de tráfico, así como realizar diferenciación de flujos mediante clases de servicio (CoS frente a las redes con enrutamiento IP tradicional. En aras de aprovechar cualidades estratégicas durante la etapa de coexistencia entre IPv4 e IPv6 existen 4 métodos para proveer conectividad a islas IPv6 [1] remotas a través de una infraestructura de core MPLS con IPv4 nativo [2], sin embargo una de las formas que permite un rápida y fácil provisión de la misma dados los mínimos requisitos de configuración y de equipos es la de disponer túneles IPv6 en los enrutadores de acceso (CE de la red. No obstante, sus cuatro variantes (manual, GRE, 6to4 e IPv6 compatible IPv4 [3] resultan adecuadas o no según las características inherentes de la red a interconectar; por tanto este artículo presenta las ventajas y desventajas propias de la utilización de cada técnica de entunelamiento como resultado de la interconexión con los cuatro tipos de túneles de una red emulada mediante GNS3+Dynamips.

  6. Animal Surgery and Resources Core

    Data.gov (United States)

    Federal Laboratory Consortium — The ASR services for NHLBI research animals include: animal model development, surgery, surgical support, post-operative care as well as technical services such as...

  7. Animal Control on Wapanocca NWR

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — Animal Control Management Plan for Wapanocca NWR and newspaper articles, and other correspondence regarding managing nuisance animals at the Refuge.

  8. High throughput detection of Coxiella burnetii by real-time PCR with internal control system and automated DNA preparation

    Directory of Open Access Journals (Sweden)

    Kramme Stefanie

    2008-05-01

    Full Text Available Abstract Background Coxiella burnetii is the causative agent of Q-fever, a widespread zoonosis. Due to its high environmental stability and infectivity it is regarded as a category B biological weapon agent. In domestic animals infection remains either asymptomatic or presents as infertility or abortion. Clinical presentation in humans can range from mild flu-like illness to acute pneumonia and hepatitis. Endocarditis represents the most common form of chronic Q-fever. In humans serology is the gold standard for diagnosis but is inadequate for early case detection. In order to serve as a diagnostic tool in an eventual biological weapon attack or in local epidemics we developed a real-time 5'nuclease based PCR assay with an internal control system. To facilitate high-throughput an automated extraction procedure was evaluated. Results To determine the minimum number of copies that are detectable at 95% chance probit analysis was used. Limit of detection in blood was 2,881 copies/ml [95%CI, 2,188–4,745 copies/ml] with a manual extraction procedure and 4,235 copies/ml [95%CI, 3,143–7,428 copies/ml] with a fully automated extraction procedure, respectively. To demonstrate clinical application a total of 72 specimens of animal origin were compared with respect to manual and automated extraction. A strong correlation between both methods was observed rendering both methods suitable. Testing of 247 follow up specimens of animal origin from a local Q-fever epidemic rendered real-time PCR more sensitive than conventional PCR. Conclusion A sensitive and thoroughly evaluated real-time PCR was established. Its high-throughput mode may show a useful approach to rapidly screen samples in local outbreaks for other organisms relevant for humans or animals. Compared to a conventional PCR assay sensitivity of real-time PCR was higher after testing samples from a local Q-fever outbreak.

  9. Implementation and validation of a sensitive PCR detection method in the eradication campaign against Aleutian mink disease virus

    DEFF Research Database (Denmark)

    Jensen, Trine Hammer; Christensen, Laurids Siig; Chriél, Mariann

    2011-01-01

    of Denmark. However, re-infections and new introductions of virus into farms require a confirmatory virological test to verify the positive test results of single animals and ultimately to investigate disease transmission. A one step PCR amplifying a 374-base fragment of the NS1 gene of AMDV was compared...... to the counter-current immune electrophoresis (CIE) routinely used in the serological screening programme. Mink organs (n = 299) obtained from 55 recently infected farms and 8 non-infected farms from 2008 to 2010 were tested by PCR, and the results were found to have a high correlation with the serological...... status of the mink. The relative diagnostic sensitivity of the PCR was 94.7%, and the relative diagnostic specificity was 97.9% when read in parallel with the CIE. PCR positive samples were sequenced and phylogenetic analysis revealed high similarity within the analysed AMDV strains and to AMDV strains...

  10. Sampling and Pooling Methods for Capturing Herd Level Antibiotic Resistance in Swine Feces using qPCR and CFU Approaches

    DEFF Research Database (Denmark)

    Schmidt, Gunilla Veslemøy; Mellerup, Anders; Christiansen, Lasse Engbo;

    2015-01-01

    for commonly detected antibiotic resistance genes were developed, and used to quantify antibiotic resistance genes in total DNA from swine fecal samples that were obtained using different sampling and pooling methods. In parallel, the number of antibiotic resistant coliform indicator bacteria was determined...... in the same swine fecal samples. The results showed that the qPCR assays were capable of detecting differences in antibiotic resistance levels in individual animals that the coliform bacteria colony forming units (CFU) could not. Also, the qPCR assays more accurately quantified antibiotic resistance genes......The aim of this article was to define the sampling level and method combination that captures antibiotic resistance at pig herd level utilizing qPCR antibiotic resistance gene quantification and culture-based quantification of antibiotic resistant coliform indicator bacteria. Fourteen qPCR assays...

  11. Validation and aplication of a polymerase chain reaction (PCR to detect porcine circovirus type 2 (PCV-2 in swine sera

    Directory of Open Access Journals (Sweden)

    Luisa Fernanda Villadiego Marmolejo

    2007-12-01

    Full Text Available Porcine circovirosis is an infectious-contagious syndrome caused by porcine circovirus type 2 (PCV-2 found mainly in recently weaned piglets causing dermatitis, neurological and reproductive disorders, pneumonia and encephalitis. The objectives of the present study were to validate a Polymerase Chain Reaction (PCR technique to detect PCV-2 in swine serum and to apply the validated technique in swine serum samples to detect PCV-2. After the application of two different PCRs, 100% of the surveyed animals were negative to PCV-2; furthermore, the PCR targeted to a region between ORFs 1 and 2 of the virus was found more sensitive when compared to another PCR targeted to the capsid protein gene. As a conclusion, PCR is a valid technique to detect PCV-2 in swine serum and the surveyed population was free of the virus.

  12. Animating facial images with drawings

    OpenAIRE

    Tunali, Gamze Dilek

    1996-01-01

    Ankara : Bilkent Univ., 1996. Thesis (Master's) -- Bilkent University, 1996. Includes bibliographical references leaves 54-56. The work presented here describes the power of 2D animation with texture mai^ping controlled by line drawings. Animation is specifically intended for facial animation and not restricted by the human face. We initially have a sequence of facial images which are taken from a video sequence of the same face and an image of another face to be animated...

  13. Directed animals in the gas

    OpenAIRE

    Borgne, Yvan Le; Marckert, Jean-François

    2006-01-01

    In this paper, we revisit the enumeration of directed animals using gas models. We show that there exists a natural construction of random directed animals on any directed graph together with a particle system that explains at the level of objects the formal link known between the density of the gas model and the generating function of directed animals counted according to the area. This provides some new methods to compute the generating function of directed animals counted according to area...

  14. Professor: The Animal Planet Optimization

    OpenAIRE

    Satish Gajawada

    2014-01-01

    This paper is dedicated to everyone who is interested in making this planet a better place to live. In the past, researchers have explored behavior of several animals separately. But there is scope to explore in the direction where various artificial animals together solve the optimization problem. In this paper, Satish Gajawada proposed The AnimalPlanet Optimization. The concept of this paper is to imitate all the animals on this planet. The idea is to solve the optimization problem where al...

  15. Rapid PCR thermocycling using microscale thermal convection.

    Science.gov (United States)

    Muddu, Radha; Hassan, Yassin A; Ugaz, Victor M

    2011-03-05

    Many molecular biology assays depend in some way on the polymerase chain reaction (PCR) to amplify an initially dilute target DNA sample to a detectable concentration level. But the design of conventional PCR thermocycling hardware, predominantly based on massive metal heating blocks whose temperature is regulated by thermoelectric heaters, severely limits the achievable reaction speed(1). Considerable electrical power is also required to repeatedly heat and cool the reagent mixture, limiting the ability to deploy these instruments in a portable format. Thermal convection has emerged as a promising alternative thermocycling approach that has the potential to overcome these limitations(2-9). Convective flows are an everyday occurrence in a diverse array of settings ranging from the Earth's atmosphere, oceans, and interior, to decorative and colorful lava lamps. Fluid motion is initiated in the same way in each case: a buoyancy driven instability arises when a confined volume of fluid is subjected to a spatial temperature gradient. These same phenomena offer an attractive way to perform PCR thermocycling. By applying a static temperature gradient across an appropriately designed reactor geometry, a continuous circulatory flow can be established that will repeatedly transport PCR reagents through temperature zones associated with the denaturing, annealing, and extension stages of the reaction (Figure 1). Thermocycling can therefore be actuated in a pseudo-isothermal manner by simply holding two opposing surfaces at fixed temperatures, completely eliminating the need to repeatedly heat and cool the instrument. One of the main challenges facing design of convective thermocyclers is the need to precisely control the spatial velocity and temperature distributions within the reactor to ensure that the reagents sequentially occupy the correct temperature zones for a sufficient period of time(10,11). Here we describe results of our efforts to probe the full 3-D velocity and

  16. AGATE animation - business theme

    Science.gov (United States)

    1998-01-01

    Business jet 2 of 6. Advanced General Aviation Technology Experiment (AGATE). Few objects convey wealth and power like a private airplane, but one day you won't have to be rich or famous to fly one. NASA is working with industry and other government agencies to develop the technology and vision for business and personal travel of the future. It's a future in which travelers fly to their destinations in small, safe, affordable and easy-to-use jets out of 'smart airports.' Future small aircraft may cost about as much as a luxury automobile. They will use 25% less fuel than today's airplanes with fuel efficiencies rivaling automobiles, but at four times highway speeds. The goal is to put 'wings on America' and enable doorstep-to-destination travel at four times the speed of highways, making it possible to go where you want, when you want - faster than ever. Image from AGATE 'business jet' video animation.

  17. Animating the Ethical Demand

    DEFF Research Database (Denmark)

    Vistisen, Peter; Jensen, Thessa; Poulsen, Søren Bolvig

    2015-01-01

    This paper addresses the challenge of attaining ethical user stances during the design process of products and services and proposes animation-based sketching as a design method, which supports elaborating and examining different ethical stances towards the user. The discussion is qualified...... dispositions, as well as create an incentive for ethical conduct in development and innovation processes. The ethical fulcrum evolves around Løgstrup’s Ethical Demand and his notion of spontaneous life manifestations. From this, three ethical stances are developed; apathy, sympathy and empathy. By exploring...... both apathetic and sympathetic views, the ethical reflections are more nuanced as a result of actually seeing the user experience simulated through different user dispositions. Exploring the three ethical stances by visualising real use cases with the technologies simulated as already being implemented...

  18. Animal models of narcolepsy.

    Science.gov (United States)

    Chen, Lichao; Brown, Ritchie E; McKenna, James T; McCarley, Robert W

    2009-08-01

    Narcolepsy is a debilitating sleep disorder with excessive daytime sleepiness and cataplexy as its two major symptoms. Although this disease was first described about one century ago, an animal model was not available until the 1970s. With the establishment of the Stanford canine narcolepsy colony, researchers were able to conduct multiple neurochemical studies to explore the pathophysiology of this disease. It was concluded that there was an imbalance between monoaminergic and cholinergic systems in canine narcolepsy. In 1999, two independent studies revealed that orexin neurotransmission deficiency was pivotal to the development of narcolepsy with cataplexy. This scientific leap fueled the generation of several genetically engineered mouse and rat models of narcolepsy. To facilitate further research, it is imperative that researchers reach a consensus concerning the evaluation of narcoleptic behavioral and EEG phenomenology in these models.

  19. Current status of animal welfare and animal rights in China.

    Science.gov (United States)

    Lu, Jiaqi; Bayne, Kathryn; Wang, Jianfei

    2013-11-01

    In the past few years, new social passions have sparked on the Chinese mainland. At the centre of these burgeoning passions is a focus on animal welfare, animal treatment, and even animal rights, by the public and academic sectors. With China's rapid economic changes and greater access to information from around the world, societal awareness of animal issues is rising very fast. Hastening this paradigm shift were several highly public incidents involving animal cruelty, including exposés on bear bile harvesting for traditional Chinese medicine, the thousands of dogs rescued from China's meat trade, and the call to boycott shark fin soup and bird nest soup. This article outlines the current status of campaigning by animal advocates in China (specifically the animal rights movement) from three interlinked perspectives: wildlife conservation, companion animal protection, and laboratory animal protection. By reviewing this campaigning, we attempt to present not only the political and social impact of the concept of animal rights, but also the perceptions of, and challenges to, animal rights activities in China.

  20. A quadruplex PCR (qxPCR) assay for adulteration in dairy products.

    Science.gov (United States)

    Agrimonti, Caterina; Pirondini, Andrea; Marmiroli, Marta; Marmiroli, Nelson

    2015-11-15

    This study describes the development of a quadruplex quantitative Real Time PCR (qxPCR) based on SYBR®GreenER chemistry, for rapid identification of DNA of cow, goat, sheep and buffalo in dairy products, and for quantification of cow DNA in these products. The platform was applied to: (i) mixes of milks at fixed percentages; (ii) cheeses prepared with the same mixes; (iii) commercial dairy products. The methodology enabled the detection of DNA from cow in mixes of milk and cheeses with a limit of detection (LOD) of 0.1%. When applied to commercial dairy products the qxPCR gave results comparable with each single-plex Real Time PCR. A good correlation (R(2)>0.9) between peaks' area of derivative of melting curves of amplicons and percentages of cow milk in milk mixes and cheeses, allows for an estimation of cow DNA in a dynamic range varying from 0.1-5% to 1-25%.

  1. The Animal Without A Head

    Institute of Scientific and Technical Information of China (English)

    万钧

    2002-01-01

    Have you ever seen an animal with out a head?there is such an animal! it has no tail or legs ,its body is full of holes it eats and breathes but never moves,it lives under water,the water brings the animal air.

  2. The Tree of Animal Life

    Science.gov (United States)

    Braude, Stan

    2007-01-01

    In this article, the author describes a short activity which introduces third- to fifth-grade students to animal classification. The Tree of Animal Life activity is a simple, sorting exercise that can help them see a bigger picture. The activity sets the stage for learning about animal taxonomy and introduces the characteristics of various animal…

  3. Animals in life and works

    Institute of Scientific and Technical Information of China (English)

    张振娟

    2014-01-01

    Many learners have made researches on animal words. Because animals have been keeping a good relationship with human beings since the human were born. So animals are important in daily life, they are also used in many works to express the Author’s thoughts.

  4. Clay Animals and Their Habitats

    Science.gov (United States)

    Adamson, Kay

    2010-01-01

    Creating clay animals and their habitats with second-grade students has long been one of the author's favorite classroom activities. Students love working with clay and they also enjoy drawing animal homes. In this article, the author describes how the students created a diorama instead of drawing their clay animal's habitat. This gave students…

  5. Animals in Environmental Education Research

    Science.gov (United States)

    Spannring, Reingard

    2017-01-01

    Over the past few decades, the increase in public and scholarly attention to human-animal relations has inspired an animal turn in a number of academic disciplines including environmental education research. This paper reviews the literature on animals in environmental education with respect to its theoretical foundations in critical pedagogy,…

  6. 9 CFR 79.4 - Designation of scrapie-positive animals, high-risk animals, exposed animals, suspect animals...

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Designation of scrapie-positive... (INCLUDING POULTRY) AND ANIMAL PRODUCTS SCRAPIE IN SHEEP AND GOATS § 79.4 Designation of scrapie-positive..., noncompliant flocks, and source flocks; notice to owners. (a) Designation. A designated scrapie...

  7. Oxygen and Early Animal Evolution

    Science.gov (United States)

    Xiao, S.

    2012-12-01

    It is often hypothesized that the rise of animals was triggered by an increase in O2 levels in the atmosphere and oceans. However, this hypothesis is remarkably difficult to test, because the timing of animal divergences is poorly resolved, the physiology of early animals is often unknown, estimates of past pO2 levels come with large error bars, and causal relationships between oxygenation and animal evolution are difficult to establish. Nonetheless, existing phylogenetic, paleontological, and geochemical data indicate that the evolution of macroscopic animals and motile macrometazoans with energetically expensive lifestyles may be temporally coupled with ocean oxygenation events in the Ediacaran Period. Thus, it is plausible that ocean oxygenation may have been a limiting factor in the early evolution of macroscopic, complex, and metabolically aggressive animals (particularly bilaterian animals). However, ocean oxygenation and animal evolution were likely engaged in two-way interactions: Ediacaran oxygenation may have initially lifted a physiological barrier for the evolution of animal size, motility, and active lifestyles, but subsequent animal diversification in the Paleozoic may have also changed oceanic redox structures. Viewed in a broader context, the early evolutionary history of animals was contingent upon a series of events, including genetic preparation (developmental genetics), environmental facilitation (oceanic oxygenation), and ecological escalation (Cambrian explosion), but the rise of animals to ecological importance also had important geobiological impacts on oceanic redox structures, sedimentary fabrics, and global geochemical cycles.

  8. Detection of Brucella spp. in bottlenose dolphins Tursiops truncatus by a real-time PCR using blowhole swabs.

    Science.gov (United States)

    Wu, Qingzhong; Conway, Jessica; Phillips, Kristen M; Stolen, Megan; Durden, Wendy N; Fauquier, Deborah; McFee, Wayne E; Schwacke, Lori

    2016-08-09

    Blowhole swabs are a simple and non-invasive method for collecting samples from cetaceans and can be used for screening large numbers of animals in the field. This study reports a real-time PCR assay for the detection of Brucella spp. using blowhole swab samples from bottlenose dolphins Tursiops truncatus stranded in the coastal region of Virginia, South Carolina and northern Florida, USA, between 2013 and 2015. We used real-time PCR results on lung samples from the same dolphins in order to estimate the relative sensitivity and specificity of real-time PCR of blowhole swabs. Brucella DNA was detected in lung tissue of 22% (18/81) and in blowhole swabs of 21% (17/81) of the sampled dolphins. The relative sensitivity and specificity of real-time PCR on blowhole swabs as compared to the real-time PCR on lung samples was 94% (17/18) and 100% (63/63), respectively. These results indicate that real-time PCR on blowhole swabs may be used as a non-invasive test for rapid detection of Brucella spp. in the respiratory tract of dolphins. To our knowledge, this is the first report on the use of blowhole swabs for detection of bacterial pathogens by real-time PCR in bottlenose dolphins.

  9. 9 CFR 95.20 - Animal manure.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Animal manure. 95.20 Section 95.20 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE EXPORTATION AND IMPORTATION OF ANIMALS (INCLUDING POULTRY) AND ANIMAL PRODUCTS SANITARY CONTROL OF...

  10. Chicken QTL mapping by multiplex PCR

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    To facilitate rapid determination of the chromosomal location of quantitative trait loci, the current approaches to gene mapping are improved using a multiplex PCR technique. The high-throughput linkage analysis method described here allows selection of 178 from 328 microsatellite markers through the multiplex PCR method combined with the semi-automatic fluorescence-labeled DNA analysis technology. Those polymorphism markers are distributed on 23 autosomes and one sex chromosome (chromosome Z), covering 3080cM genetic distance. The average marker density is 18cM, dispersed into 30 different sets. These selected polymorphism microsatellite markers segregate with the family members, following the Mendel's heritage laws, and are very useful for chicken linkage map analysis as well as for the research on some important economic quantitative characters of chicken.

  11. PCR-SSCP检测水稻SNPs

    Institute of Scientific and Technical Information of China (English)

    李飞; 郭凌; 余潮

    2006-01-01

    本文选择5个水稻SNPs位点为对象,研究凝胶组成和电泳条件等对PCR-SSCP检测SNPs的影响.发现凝胶浓度、交联度、甘油和电泳温度等均可明显影响DNA单链分子迁移率,且对不同DNA单链分子的影响程度不同.因此,可以通过调节凝胶浓度、交联度、甘油和电泳温度等提高PCR-SSCP检测水稻SNPs的分辨率及效果.在一定条件下,较低交联度和4°电泳条件检测SNPs的效果更好.

  12. IDENTIFICATION OF ANIMAL ADHESIVES USING DNA AMPLIFICATION

    DEFF Research Database (Denmark)

    Eriksen, Anne Marie; Kristensen, Hans Viborg; Bøllingtoft, Peder

    2014-01-01

    The aim of this study was to examine whether DNA was degraded in the manufacturing of animal glue. To test this, two different types of sturgeon glue (Acipenser sp.) were manufactured using historic recipes. One glue was boiled for a substantial amount of time and the other was kept under 75°C. DNA...... samples were collected from both glues in order to test whether the DNA was degraded in the heating process of making the glue. It was also tested whether two different kinds of flex canvas (for paintings), one coarse and one fine weaved would inhibit the PCR reaction. To do this the glue were applied...... of the glue and in 18 out of 24 samples collected of the canvas. In four out of the five cases where it was not possible amplify DNA, the sample belongs to the smallest size of the canvas investigated. As shown in this study it is possible to get DNA out of boiled animal glue and glue applied onto canvas...

  13. Identification of canine papillomavirus by PCR in Greyhound dogs

    Directory of Open Access Journals (Sweden)

    Eman A. Anis

    2016-12-01

    Full Text Available Background Corns are hard protuberances that occur on the digital footpads of Greyhound dogs. The cause of these lesions is unknown and there is little information about them in the veterinary literature. We received anecdotal examples of dog to dog spread of corns suggesting an infectious cause. The aim of this study was to determine if papillomavirus (PV is associated with Greyhound corns. Methods We examined four corns from two unrelated adult Greyhound dogs that resided in Florida and Washington, respectively, for PV by PCR. The samples were obtained by owner coring of two lesions from one dog and laser removal of two lesions from the other dog. Total nucleic acid was extracted and DNA was amplified using two PCR primer sets that have been shown to amplify a broad range of PVs from humans and animals: FAP59/ FAP64 and MY11/ MY09. The DNA sequences were compared with all sequences in GenBank. Formalin-fixed, paraffin-embedded tissue from the footpads of four dogs with other inflammatory dermatoses were also examined. Results PV DNA was amplified from all four corn lesions, while no PV DNA was amplified from other tissues. Comparison of the 444-bp sequences amplified by the MY11/ MY09 primers identified two different PVs. One showed 96% nucleotide sequence similarity with the L1 gene of canine PV type 12. The other showed 78% similarity to canine PV type 16 and, therefore, represents a novel PV. In one of the corns, infection by two of the identified PVs was found. Discussion These results suggest PV infection could be involved in the pathogenesis of corns in Greyhound dogs.

  14. Obtención de electricidad y agua potable mediante energía solar para un centro de salud mozambiqueño

    OpenAIRE

    Antona Díaz, Ana María

    2009-01-01

    El proyecto consiste en cubrir las necesidades de energía eléctrica y agua potable del centro de salud de Minehuene (Mozambique), mediante energía solar fotovoltaica y térmica. La cobertura de la demanda de electricidad del centro se realiza mediante un sistema de energía fotovoltaico. Para la obtención de agua potable se evalúan diferentes alternativas: pasteurización del agua de lluvia mediante paneles solares térmicos, purificación de esta agua mediante lámparas ultravioletas u obtenci...

  15. Application of PCR techniques in toxicology

    Directory of Open Access Journals (Sweden)

    Maja Kazubek

    2010-10-01

    Full Text Available Molecular biology techniques have become widely used in toxicology, leading to the creation of a new science – molecular toxicology. The goal of molecular toxicology is to detect and study the changes induced by xenobiotics at the molecular level. The research scope of molecular toxicology includes examination of mutations in genomic DNA, differences in mRNA expression and study of genotype indicating individual sensitivity.The processes of activation and detoxification of xenobiotics, drugs and environmental carcinogens involve several enzymes (xenobiotic-metabolizing enzymes – XMEs. Most of the chemicals entering our bodies, regardless of whether they have medical, pathogenic or carcinogenic properties, require metabolic activation by phase I enzymes (cytochrome P-450. In the next process the phase I products are usually detoxified by phase II enzymes, mainly by epoxide hydrolase, glutathione transferase, N-acetyltransferase or sulfotransferase. PCR techniques allow precise study of the effects of xenobiotics on cells and tissues by examining the level of activation of genes coding for phase I and II enzymes, or by testing the activity of other elements of the transcriptome. Studies of sensitivity of individual cells or tissues based on examination of mutation or gene polymorphism presence are also relevant.This paper presents the possibility of using various PCR techniques in toxicology and especially in the study of genetically determined sensitivity to xenobiotics. It also covers the possibilities of applying qPCR and qRT-PCR methods in the search for exposure biomarkers with particular emphasis on individual cytochrome P450 isoforms. Furthermore, it provides information about the possibility of implementing the differential display technique in the identification of new genes activated by toxic agents.

  16. Reproducibility of AMPLICOR enterovirus PCR test results.

    OpenAIRE

    1997-01-01

    The reproducibility of AMPLICOR enterovirus PCR test results was determined with clinical samples of cerebrospinal fluid, serum, urine, and throat and rectal swabs. Among 608 samples from which duplicate aliquots were run simultaneously, only seven pairs gave discordant results. Among 104 samples from which duplicate aliquots were run in separate assays, no discordance was seen. Overall, the reproducibility of test kit results was 99% (705 of 712).

  17. Comparison of Droplet Digital PCR and Quantitative PCR Assays for Quantitative Detection of Xanthomonas citri Subsp. citri.

    Directory of Open Access Journals (Sweden)

    Yun Zhao

    Full Text Available Droplet digital polymerase chain reaction (ddPCR is a novel molecular biology technique providing absolute quantification of target nucleic acids without the need for an external calibrator. Despite its emerging applications in medical diagnosis, there are few reports of its use for the detection of plant pathogens. This work was designed to assess the diagnosis potential of the ddPCR for absolute quantitative detection of Xanthomonas citri subsp. citri, a quarantine plant pathogenic bacterium that causes citrus bacterial canker in susceptible Citrus species. We transferred an established quantitative PCR (qPCR assay for citrus bacterial canker diagnosis directly to the ddPCR format and compared the performance of the two methods. The qPCR assay has a broader dynamic range compared to the ddPCR assay and the ddPCR assay has a significantly higher degree of sensitivity compared to the qPCR assay. The influence of PCR inhibitors can be reduced considerably in the ddPCR assay because the collection of end-point fluorescent signals and the counting of binomial events (positive or negative droplets are associated with a Poisson algorithm. The ddPCR assay also shows lower coefficient of variation compared to the qPCR assay especially in low target concentration. The linear association of the measurements by ddPCR and qPCR assays is strong (Pearson correlation = 0.8633; P<0.001. Receiver operating characteristic analysis indicates the ddPCR methodology is a more robust approach for diagnosis of citrus bacterial canker. In summary, the results demonstrated that the ddPCR assay has the potential for the quantitative detection of X. citri subsp. citri with high precision and accuracy as compared with the results from qPCR assay. Further studies are required to evaluate and validate the value of ddPCR technology in the diagnosis of plant disease and quarantine applications.

  18. Comparison of noninvasive sample collection procedures for canine leishmaniasis diagnosis by PCR-hybridization

    Energy Technology Data Exchange (ETDEWEB)

    Ferreira, Sidney de Almeida; Andrade, Antero Silva Ribeiro de [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil)]. E-mail: vidasnino@yahoo.com.br; antero@cdtn.br; Ituassu, Leonardo Trindade; Melo, Maria Norma de [Universidade Federal de Minas Gerais, Belo Horizonte, MG (Brazil)]. E-mail: melo@mono.icb.ufmg.br; ltituassu@yahoo.com.br

    2007-07-01

    The dogs are the main reservoir of the visceral leishmaniasis etiological agent Leishmania chagasi and these animals have to be systematically monitored. The aim of present work was to standardize a method for canine leishmaniasis diagnosis using DNA samples obtained by a noninvasive ways. Two kind of samples were compared: conjunctival swab and blood. The samples were analyzed by the Polymerase Chain Reaction (PCR) associated with the hybridization of {sup 32}P labeled DNA probes. An in vitro test was carried out using cotton swabs seeded with L. chagasi parasites at different cell numbers. After that, the PCR and hybridization sensitivity was evaluated in two groups of 23 seropositive dogs. Conjunctival swabs and 1,0 mL of blood were collected from each animal. 90 {mu}L of these blood were spotted onto filter paper and the remaining used to prepare the buffy coat. The DNA purification from cotton swabs was carried out through the phenol-chloroform (group 1) or boiling (group 2). The Wizard kit was used to DNA extraction from buffy coat. The filters were treated according to Dialab protocol. The analysis of the seeded samples showed that the PCR was able to identify until ten parasites while the following hybridization of the PCR products allows the detection of until one parasite. The PCR positivity for the conjunctival swabs were 73.9% and 52.2% respectively to the groups 1 and 2. For buffy coat the positivities were 43.5% and 56.5% respectively. The filters presented the lowest positivity. The hybridization step was not accomplished yet for these samples. (author)

  19. Why not look at animals?

    Directory of Open Access Journals (Sweden)

    Anat Pick

    2015-04-01

    Full Text Available Revisiting John Berger’s seminal essay ‘Why Look at Animals?’ (1980, this essay inverts Berger’s title in order to explore instances where the visibility of animals is at stake and where seeing is linked to forms of surveillance and control. In the context of advanced optical and tracking technologies that render animals permanently visible, the possibility of not-seeing emerges as a progressive modality of relation to animals that takes seriously the notion of animal privacy and the exposed animal’s resistance to the human gaze.

  20. Digital Animation Character Creation Design

    Institute of Scientific and Technical Information of China (English)

    潘锋

    2014-01-01

    The purpose of this article is to discuss the proper method for Chinese digital animation character design on the foundation of certain cultural elements. The method used in this study is known as comparative analysis of Disney and Japanese animation styles in action, appearance, facial expression and voice design. These dynamic factors are the best carrier of the animation spirit and native culture, so it is important to take the dynamic factors into account when producing the digital animation, and it will be an excellent starting point to innovate Chinese digital animation.

  1. Amp-PCR: combining a random unbiased Phi29-amplification with a specific real-time PCR, performed in one tube to increase PCR sensitivity.

    Directory of Open Access Journals (Sweden)

    Lena Erlandsson

    Full Text Available In clinical situations where a diagnostic real-time PCR assay is not sensitive enough, leading to low or falsely negative results, or where detection earlier in a disease progression would benefit the patient, an unbiased pre-amplification prior to the real-time PCR could be beneficial. In Amp-PCR, an unbiased random Phi29 pre-amplification is combined with a specific real-time PCR reaction. The two reactions are separated physically by a wax-layer (AmpliWax® and are run in sequel in the same sealed tube. Amp-PCR can increase the specific PCR signal at least 100×10(6-fold and make it possible to detect positive samples normally under the detection limit of the specific real-time PCR. The risk of contamination is eliminated and Amp-PCR could replace nested-PCR in situations where increased sensitivity is needed e.g. in routine PCR diagnostic analysis. We show Amp-PCR to work on clinical samples containing circular and linear viral dsDNA genomes, but can work well on DNA of any origin, both from non-cellular (virus and cellular sources (bacteria, archae, eukaryotes.

  2. Clostridium perfringens isolate typing by multiplex PCR

    Directory of Open Access Journals (Sweden)

    MR Ahsani

    2010-01-01

    Full Text Available 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.

  3. Animal Welfare in Air Transport

    Directory of Open Access Journals (Sweden)

    Boris Popović

    2012-10-01

    Full Text Available Animal welfare is becoming an evermore-important factorfor air carriers from the economical viewpoint, due to its importantimpact on the carrier public image. High standard care hasto be taken of animals during transport in order to satisfy an importantsegment of airline customers, either the Business/Firstclass passengers travelling with pets, or influential shippers ofracing horses, dogs, Zoo species etc.Air transp011 of animals, disregarding other advantages,may pose a threat to their health and welfare being a significantmultifactorial stressor. Along with cardiovascular, endocrineand metabolic abe1mtions, it affects the immune response ofan animal and increases susceptibility to infection. Therefore,strict conditions for air transport of eve1y animal species havebeen imposed. Transport of only healthy animals is approved,as it is necessG/y to prevent the spread of disease during transportand to provide satisfactOJy environment for animals to betransported.

  4. Knowledge of the animal welfare act and animal welfare regulations influences attitudes toward animal research.

    Science.gov (United States)

    Metzger, Mitchell M

    2015-01-01

    Recent public-opinion polls indicate that Americans have shown a decline in support for animal experimentation, and several reports suggest a relationship between people's knowledge of animal welfare regulations and their attitudes toward animal research. Therefore, this study was designed to assess respondent's knowledge of several provisions in the Animal Welfare Act (AWA) and Animal Welfare Regulations (AWR), and determine whether exposure to elements of this legislation would influence an individual's attitudes toward the use of animals in research. A survey was used to assess knowledge of animal research regulations and attitudes toward animal research from a sample of individuals recruited through Amazon's Mechanical Turk crowdsourcing marketplace. Results from study 1 confirmed the hypothesis that respondents had little knowledge of various federal regulations that govern animal research activities. Data from study 2 revealed that exposure to elements of the AWA and AWR influenced participants' attitudes toward the use of animals in research. These results suggest that providing information to the general public about the AWA and AWR that protect laboratory animals from abuse and neglect may help alleviate concerns about using animals in research settings.

  5. Control de Copitarsia decolora en cultivos de flores mediante la emisión de frecuencias

    Directory of Open Access Journals (Sweden)

    Hernán Paz

    2008-05-01

    Full Text Available Una de las plagas interceptadas por el Servicio de Inspección de Salud de Animales y Plantas (APHIS de Estados Unidos, en las exportaciones de algunas especies de flores de corte de Colombia, es Copitarsia spp. El presente estudio evaluó la respuesta evasiva de Copitarsia decolora (Lepidoptera: Noctuidae mediante la simulación de llamadas de ecolocación de murciélagos como propuestapara el manejo en salas de poscosecha de fl ores, bajo condiciones controladas. Las formas de las señales de ecolocación de los murciélagos se tomaron de algunasmuestras de quirópteros insectívoros y se simularon por medio del software "Sound Ruler", determinando los parámetros fundamentales que caracterizan las llamadas, como amplitud, frecuencia y forma, entre otros. A partir de esta caracterización, se diseñó y construyó un sistema electrónico capaz de reproducir las llamadas de ecolocación. Se probó el sistema, emitiendo la señal reproducida hacia una muestra de Copitarsia, y se observó una reacción evasiva frente a dicha señal./ One of the plagues intercepted by the Health's supervisory Service of Animals and Plants (APHIS of United States in the exports of some species of fl owers of court of Colombia is Copitarsia spp. The present study evaluated the evasive answer of Copitarsia decolora (Lepidoptera: Noctuidae by means of the simulation of calls of echolocation of bats like proposal for the handling in rooms of post-harvest of flowers, under controlled conditions. The shapes of the signs of echo-location of the bats took of some samples of insectivorous quirópteros and were simulated by means of the software "Sound Ruler", determining the fundamental parameters that characterize the calls as their width, frequency and it forms, between other ones; starting from this characterization, it was designed and built an electronic system able to reproduce the echo-location calls. The system was tried, emitting the sign reproduced toward a

  6. Molecular characterization of Blastocystis isolates from zoo animals and their animal-keepers.

    Science.gov (United States)

    Parkar, Unaiza; Traub, Rebecca J; Vitali, Simone; Elliot, Aileen; Levecke, Bruno; Robertson, Ian; Geurden, Thomas; Steele, Jan; Drake, Bev; Thompson, R C Andrew

    2010-04-19

    Blastocystis is an enteric protist and one of the most frequently reported parasitic infections in humans and a variety of animal hosts. It has also been reported in numerous parasite surveys of animals in zoological gardens and in particular in non-human primate species. PCR-based methods capable of the direct detection of Blastocystis in faeces were used to detect Blastocystis from various hosts, including non-human primates, Australian native fauna, elephants and giraffes, as well as their keepers from a Western Australian zoo. Additional faecal samples were also collected from elephants and giraffes from four other zoos in Amsterdam (The Netherlands), Antwerp (Belgium), Melbourne and Werribee (Australia). Information regarding the general health and lifestyle of the human volunteers were obtained by questionnaire. Overall, 42% and 63% of animals and zoo-keepers sampled from the Western Australian zoo were positive for Blastocystis, respectively. The occurrence of Blastocystis in elephants and giraffes from other cities was similar. This is the first report of Blastocystis found in the elephant, giraffe, quokka, southern hairy nosed wombat and western grey kangaroo. Three novel and what appear to be highly host-specific subtypes (STs) of Blastocystis in the elephant, giraffe and quokka are also described. These findings indicate that further exploration of the genetic diversity of Blastocystis is crucial. Most zoo-keepers at the Perth Zoo were harbouring Blastocystis. Four of these zoo-keeper isolates were identical to the isolates from the southern hairy nosed wombat and five primate species.

  7. Remoción de Contaminantes Atmosféricos Sox y Nox mediante Nanotubos de Carbono

    OpenAIRE

    Saavedra Labastida, Erika

    2013-01-01

    En el presente proyecto de investigación se realizó la evaluación de la remoción de los contaminantes atmosféricos provenientes de fuentes móviles, mediante nanotubos de carbono (NTC) comerciales. Para conocer las propiedades de los NTC empleados, fueron caracterizados fisicoquímicamente mediante diferentes análisis tales como MEB, MET, Raman, DRX y BET, los resultados que se obtuvieron mostraron óptimas propiedades fisicoquímicas las cuales permiten que sean empleados como ...

  8. Selección de estaciones termométricas mediante un filtro de comparación de curvas.

    OpenAIRE

    José Antonio Sosa Cardo

    2001-01-01

    A la hora de realizar cualquier estudio climatológico o interpolación espacial es fundamental contar con un conjunto de estaciones fiables, y desechar aquellas que lo único que nos van a aportar es ruido. Se han desarrollado dos métodos para seleccionar estaciones de forma objetiva: 1) Mediante análisis de las formas de las curvas de las series, siendo necesaria otra estación con una curva similar para que se de por válida una estación. 2) Mediante análisis del valor diario. Cuando existe una...

  9. In vitro culture, PCR , and nested PCR for the detection of Theileria equi in horses submitted to exercise Cultivo in vitro, PCR e nested PCR na detecção de Theileria equi em eqüinos submetidos a exercícios

    Directory of Open Access Journals (Sweden)

    C.D. Baldani

    2008-06-01

    Full Text Available This study compared the usefulness of in vitro culture, PCR, and nested PCR for the diagnosis of Theileria equi in horses submitted to stress during exercise. Blood samples from 15 apparently healthy horses, previously conditioned to a high-speed equine treadmill, were taken prior to and after exercise. The animals were divided into two experimental groups: 30-day training schedule (G1 and 90-day training schedule (G2. Statistical analysis was performed using a chi-square test and kappa statistic was used in order to assess agreement. No significant difference was observed between samples collected at resting or after exercise. In G1, merozoites of T. equi were detected in the blood smears of four horses before in vitro culture, whereas 14 samples were positive, confirmed by culture. In G2, five and 11 horses were positive before and after culture, respectively. No PCR amplified product was observed in any of the tested animals although the PCR system based on the 16S rRNA gene of T. equi detected DNA in blood with an equivalent 8x10-5% parasitaemia. The nested PCR based on the T. equi merozoite antigen gene (EMA-1 allowed the visualization of amplified products in all the horses. Therefore, nested PCR should be considered as a means of detection of sub-clinical T. equi infections and in vitro culture could be used as a complement to other methods of diagnosis.Comparou-se a utilização do cultivo in vitro, PCR e nested PCR no diagnóstico de Theileria equi em eqüinos submetidos ao estresse induzido por exercícios. Amostras de sangue foram obtidas de 15 eqüinos submetidos a treinamento em esteira rolante de alto desempenho, sendo as amostras colhidas antes e após os exercícios. Os animais foram divididos em dois grupos experimentais: 30 dias de treinamento (G1 e 90 dias de treinamento (G2. O teste do qui-quadrado foi empregado para as análises estatísticas e o índice kappa utilizado para avaliar a concordância. Não houve diferen

  10. An insulated isothermal PCR method on a field-deployable device for rapid and sensitive detection of canine parvovirus type 2 at points of need.

    Science.gov (United States)

    Wilkes, Rebecca P; Lee, Pei-Yu A; Tsai, Yun-Long; Tsai, Chuan-Fu; Chang, Hsiu-Hui; Chang, Hsiao-Fen G; Wang, Hwa-Tang T

    2015-08-01

    Canine parvovirus type 2 (CPV-2), including subtypes 2a, 2b and 2c, causes an acute enteric disease in both domestic and wild animals. Rapid and sensitive diagnosis aids effective disease management at points of need (PON). A commercially available, field-deployable and user-friendly system, designed with insulated isothermal PCR (iiPCR) technology, displays excellent sensitivity and specificity for nucleic acid detection. An iiPCR method was developed for on-site detection of all circulating CPV-2 strains. Limit of detection was determined using plasmid DNA. CPV-2a, 2b and 2c strains, a feline panleukopenia virus (FPV) strain, and nine canine pathogens were tested to evaluate assay specificity. Reaction sensitivity and performance were compared with an in-house real-time PCR using serial dilutions of a CPV-2b strain and 100 canine fecal clinical samples collected from 2010 to 2014, respectively. The 95% limit of detection of the iiPCR method was 13 copies of standard DNA and detection limits for CPV-2b DNA were equivalent for iiPCR and real-time PCR. The iiPCR reaction detected CPV-2a, 2b and 2c and FPV. Non-targeted pathogens were not detected. Test results of real-time PCR and iiPCR from 99 fecal samples agreed with each other, while one real-time PCR-positive sample tested negative by iiPCR. Therefore, excellent agreement (k = 0.98) with sensitivity of 98.41% and specificity of 100% in detecting CPV-2 in feces was found between the two methods. In conclusion, the iiPCR system has potential to serve as a useful tool for rapid and accurate PON, molecular detection of CPV-2.

  11. Animal models of candidiasis.

    Science.gov (United States)

    Clancy, Cornelius J; Cheng, Shaoji; Nguyen, Minh Hong

    2009-01-01

    Animal models are powerful tools to study the pathogenesis of diverse types of candidiasis. Murine models are particularly attractive because of cost, ease of handling, technical feasibility, and experience with their use. In this chapter, we describe methods for two of the most popular murine models of disease caused by Candida albicans. In an intravenously disseminated candidiasis (DC) model, immunocompetent mice are infected by lateral tail vein injections of a C. albicans suspension. Endpoints include mortality, tissue burdens of infection (most importantly in the kidneys, although spleens and livers are sometimes also assessed), and histopathology of infected organs. In a model of oral/esophageal candidiasis, mice are immunosuppressed with cortisone acetate and inoculated in the oral cavities using swabs saturated with a C. albicans suspension. Since mice do not die from oral candidiasis in this model, endpoints are tissue burden of infection and histopathology. The DC and oral/esophageal models are most commonly used for studies of C. albicans virulence, in which the disease-causing ability of a mutant strain is compared with an isogenic parent strain. Nevertheless, the basic techniques we describe are also applicable to models adapted to investigate other aspects of pathogenesis, such as spatiotemporal patterns of gene expression, specific aspects of host immune response and assessment of antifungal agents, immunomodulatory strategies, and vaccines.

  12. [Dangerous marine animals].

    Science.gov (United States)

    Antensteiner, G

    1999-01-01

    Sea-biological basic knowledge for divers is offered only in special lessons for advanced scuba divers. According to statistics, however, five per cent of the deadly diving accidents are caused by underwater organisms. This number could be reduced to a fraction, by correct behaviour during the dive and after an accident. The most frequent accidents with sea animals during water sports are not by unprovoked shark attacks, which cause six deaths world-wide per year on the average, but turn out with usually well camouflaged sea inhabitants, that do not attack humans, rather by their inadvertence coincidentally get in contact with it. The various defense instruments of the often small, inconspicuous organisms reach from teeth over poison stings, pricks, spines, scalpelles, nettle injections and chemical weapons up to poison arrows. Due to that variety of the maritime life, the most important representatives of its type are explained including severity level of the caused injury or contamination. Both, diagnostic position and therapy possibility are described as follows: 1. Porifera (sponge), 2. Hydrozoa (white weed, yellow flower head), Actinaria (sea anemones), 3. Conidae (cone shells), Tridocna (giant clam), octopoda (octopus), 4. Acanthaster planci (crown of thorns), Echinodea (sea urchins), Holothurioidea (sea cucumber), 5. Selachoidei (shark), Batoidei (Ray), Muraenidae (moray), Plotosidae (barbel eels), Synanciidae (stonefish), Scorpaenidae (scorpionfish), Pterois (lion fish), Sphyraena Spec. (barracuda), Balistidae (triggerfish), Ostracionidae (puffer).

  13. Sedna Orbit Animation

    Science.gov (United States)

    2004-01-01

    This animation shows the location of the newly discovered planet-like object, dubbed 'Sedna,' in relation to the rest of the solar system. Starting at the inner solar system, which includes the orbits of Mercury, Venus, Earth, and Mars (all in yellow), the view pulls away through the asteroid belt and the orbits of the outer planets beyond (green). Pluto and the distant Kuiper Belt objects are seen next until finally Sedna comes into view. As the field widens the full orbit of Sedna can be seen along with its current location. Sedna is nearing its closest approach to the Sun; its 10,000 year orbit typically takes it to far greater distances. Moving past Sedna, what was previously thought to be the inner edge of the Oort cloud appears. The Oort cloud is a spherical distribution of cold, icy bodies lying at the limits of the Sun's gravitational pull. Sedna's presence suggests that this Oort cloud is much closer than scientists believed.

  14. Epileptogenesis and companion animals.

    Science.gov (United States)

    Patterson, Edward Ned E

    2013-05-01

    Epileptogenesis is the process by which a normal brain develops into an epileptic brain. There are 3 distinct phases of epileptogenesis-the latent period before seizures occur, the occurrence of recurrent seizures, and in about 30% of patients, the development of refractory epilepsy. Understanding the basic epileptic circuit abnormalities associated with recurrent seizures via aberrations in glutamate, gamma-aminobutyric acid, and ligand- and voltage-gated ion channel activity can help the small-animal practitioner understand the mechanism of action of the antiepileptic drugs currently used for dogs and cats for new-onset and refractory epilepsy. Understanding the latest research results and theories about the pathophysiology of the latent period of epileptogenesis, where recurrent seizures have not yet developed, would help the practitioner understand possible target areas for future treatments to treat epilepsy by preventing it rather than just symptomatically preventing recurrent seizures. The current areas of focus of research on the latent period include neurodegeneration, neurogenesis, axonal sprouting, glial cell activation, invasion of inflammatory cells, angiogenesis, and subclinical alteration of ligand- and receptor-gated ion channels.

  15. CERN OVERVIEW animation

    CERN Multimedia

    Arzur Catel Torres

    2015-01-01

    This animation shows how the Large Hadron Collider (LHC) works. The film begins with an aerial view of CERN near Geneva, with outlines of the accelerator complex, including the underground Large Hadron Collider (LHC), 27-km in circumference. The positions of the four largest LHC experiments, ALICE, ATLAS, CMS and LHCb are revealed before we see protons travelling around the LHC ring. The proton source is a simple bottle of hydrogen gas. An electric field is used to strip hydrogen atoms of their electrons to yield protons. Linac 2, the first accelerator in the chain, accelerates the protons to the energy of 50 MeV. The beam is then injected into the Proton Synchrotron Booster (PSB), which accelerates the protons to 1.4 GeV, followed by the Proton Synchrotron (PS), which pushes the beam to 25 GeV. Protons are then sent to the Super Proton Synchrotron (SPS) where they are accelerated to 450 GeV. The protons are finally transferred to the two beam pipes of the LHC. The beam in one pipe circulates clockwise while ...

  16. Species-specific PCR for the Diagnosis and Determination of Antibiotic Susceptibilities of Brucella Strains Isolated from Tehran, Iran

    Science.gov (United States)

    Irajian, Gholam Reza; Masjedian Jazi, Faramarz; Mirnejad, Reza; Piranfar, Vahhab; Zahraei salehi, Taghi; Amir Mozafari, Noor; Ghaznavi-rad, Ehsanollah; Khormali, Mahmoud

    2016-01-01

    Background: Brucellosis is an endemic zoonotic disease in the Middle East. This study intended to design a uniplex PCR assay for the detection and differentiation of Brucella at the species level and determining the antibiotic susceptibility pattern of Brucella in Iran. Methods: Sixty-eight Brucella specimens (38 animal and 30 human specimens) were analyzed using PCR (using one pair of primers). Antibiotic susceptibility patterns were evaluated and compared using the E-Test and disk diffusion susceptibility test. Tigecycline susceptibility pattern was compared with other antibiotics. Results: Thirty six isolates of B. melitensis, 2 isolates of B. abortus and 1 isolate of B. suis from the 38 animal specimens, 24 isolates of B. melitensis and 6 isolates of B. abortus from the 30 human specimens were differentiated. The MIC50 values of doxycycline for human and animal specimens were 125 and 10 μg/ml, respectively, tigecycline 0.064 μg/ml for human specimens and 0.125μg/ml for animal specimens, and trimethoprim/ sulfamethoxazole and ciprofloxacin 0.065 and 0.125μg/ml, respectively, for both human and animal specimens. The highest MIC50 value of streptomycin in the human specimens was 0.5μg/ml and 1μg/ml for the animal specimens. The greatest resistance shown was to tetracycline and gentamicin, respectively. Conclusion: Uniplex PCR for the detection and differentiation of Brucella at the strain level is faster and less expensive than multiplex PCR, and the antibiotics doxycycline, rifampin, trimethoprim-sulfamethoxazole, ciprofloxacin, and ofloxacin are the most effective antibiotics for treating brucellosis. Resistance to tigecycline is increasing, and we recommend that it be used in a combination regimen. PMID:27799972

  17. An Evaluation of Quantitative PCR Assays (TaqMan® and SYBR Green for the Detection of Babesia bigemina and Babesia bovis, and a Novel Fluorescent-ITS1-PCR Capillary Electrophoresis Method for Genotyping B. bovis Isolates

    Directory of Open Access Journals (Sweden)

    Bing Zhang

    2016-09-01

    Full Text Available Babesia spp. are tick-transmitted haemoparasites causing tick fever in cattle. In Australia, economic losses to the cattle industry from tick fever are estimated at AUD$26 Million per annum. If animals recover from these infections, they become immune carriers. Here we describe a novel multiplex TaqMan qPCR targeting cytochrome b genes for the identification of Babesia spp. The assay shows high sensitivity, specificity and reproducibility, and allows quantification of parasite DNA from Babesia bovis and B. bigemina compared to standard PCR assays. A previously published cytochrome b SYBR Green qPCR was also tested in this study, showing slightly higher sensitivity than the Taqman qPCRs but requires melting curve analysis post-PCR to confirm specificity. The SYBR Green assays were further evaluated using both diagnostic submissions and vaccinated cattle (at 7, 9, 11 and 14 days post-inoculation showed that B. bigemina can be detected more frequently than B. bovis. Due to fewer circulating parasites, B. bovis detection in carrier animals requires higher DNA input. Preliminary data for a novel fluorescent PCR genotyping based on the Internal Transcribed Spacer 1 region to detect vaccine and field alleles of B. bovis are described. This assay is capable of detecting vaccine and novel field isolate alleles in a single sample.

  18. Overcoming inhibition in real-time diagnostic PCR.

    Science.gov (United States)

    Hedman, Johannes; Rådström, Peter

    2013-01-01

    PCR is an important and powerful tool in several fields, including clinical diagnostics, food analysis, and forensic analysis. In theory, PCR enables the detection of one single cell or DNA molecule. However, the presence of PCR inhibitors in the sample affects the amplification efficiency of PCR, thus lowering the detection limit, as well as the precision of sequence-specific nucleic acid quantification in real-time PCR. In order to overcome the problems caused by PCR inhibitors, all the steps leading up to DNA amplification must be optimized for the sample type in question. Sampling and sample treatment are key steps, but most of the methods currently in use were developed for conventional diagnostic methods and not for PCR. Therefore, there is a need for fast, simple, and robust sample preparation methods that take advantage of the accuracy of PCR. In addition, the thermostable DNA polymerases and buffer systems used in PCR are affected differently by inhibitors. During recent years, real-time PCR has developed considerably and is now widely used as a diagnostic tool. This technique has greatly improved the degree of automation and reduced the analysis time, but has also introduced a new set of PCR inhibitors, namely those affecting the fluorescence signal. The purpose of this chapter is to view the complexity of PCR inhibition from different angles, presenting both molecular explanations and practical ways of dealing with the problem. Although diagnostic PCR brings together scientists from different diagnostic fields, end-users have not fully exploited the potential of learning from each other. Here, we have collected knowledge from archeological analysis, clinical diagnostics, environmental analysis, food analysis, and forensic analysis. The concept of integrating sampling, sample treatment, and the chemistry of PCR, i.e., pre-PCR processing, will be addressed as a general approach to overcoming real-time PCR inhibition and producing samples optimal for PCR

  19. 猪嗜血支原体PCR及荧光定量PCR检测方法的建立和比较%Establishment and comparison of the PCR and FQ-PCR assays for detection of Mycoplasma haemosuis

    Institute of Scientific and Technical Information of China (English)

    张长莹; 张莉莉; 李玉峰; 李文良; 刘捷; 陈闻; 姜平

    2011-01-01

    To compare PCR and TaqMan real time PCR (FQ-PCR) for the detection of Mycoplasma haemosuis infection in pigs, the PCR method and FQ-PCR were set up with the primers targeting the 16S rRNA gene of M.haemosuis.Plasmid containing 16S rRNA gene was used as template to optimize the conditions of conventional PCR and FQ-PCR reactions.The results showed that these 2 assays have good sensitivity and specificity.The sensitivity of FQ-PCR was 1,000 times higher than conventional PCR.Twenty clinical samples were detected by these 2 assays, positive rate was 12/20 by conventional PCR, while 15/20 by FQ-PCR.In conclusion, these 2 assays can effectively detect the clinical infection of M.haemosuis and may provide useful tools in establishment of animal infection model.%为了解猪嗜血支原体(Mycoplasma haemosuis)对猪群的感染情况并建立该病的检测方法,本研究根据GenBank登录的M.haemosuis 16S rRNA基因序列(FJ263944)设计合成PCR引物以及荧光定量PCR(FQ-PCR)引物和探针.以含16S rRNA基因的重组质粒为模板,通过对PCR反应条件的优化,建立检测M.haemosuis的PCR和FQ-PCR检测方法.结果表明,这两种检测方法均具有较好的敏感性和特异性,与常规方法相比,FQ-PCR方法的敏感性高1000倍;用这两种PCR方法检测20份临床样品,常规PCR方法的阳性率为60%(12/20),而FQ-PCR方法的阳性率为75%(15/20).这两种检测方法的建立为确定M.haemosuis在我国猪群的感染情况和建立该病动物模型提供有效的检测手段.

  20. Detection and Comparison of Pathogen of Virus Disease in Pumpkin by RT-PCR and IC-PCR

    Institute of Scientific and Technical Information of China (English)

    YANG Guohui; ZHANG Zhongkai; CUI Chongshi

    2006-01-01

    Two kinds of methods RT-PCR and IC-PCR were used to detect pathogen of virus disease of pumpkin and the sensitivity of the two methods was compared. The results showed that PRSV-W and CMV were detected in diseased samples gathered in Yunnan Province, while WMV and CMV were detected in diseased samples gathered in Heilongjiang Province. The sensitivity of RT-PCR is higher than that of IC-PCR, but the effect of IC-PCR in the specialization of bonding reaction and requisition for experiment material is better than that of RT-PCR.