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Sample records for animal mediante pcr

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

    Directory of Open Access Journals (Sweden)

    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.

  2. Localizzazione e valutazione dell’espressione di Chlamydophila pneumoniae mediante RT-PCR in situ

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    Stefania Cazzavillan

    2005-12-01

    Full Text Available Chlamydophila pneumoniae, an obligate intracellular gram negative bacterium, is involved in a wide spectrum of symptomatic respiratory tract diseases. However more recently it has been reported to be a pathogenic agent in the mechanism leading to atherosclerosis. In the present study the presence of Chlamydophila pneumoniae was assessed, using nested PCR and in situ PCR, while the viability of the microorganism was investigated using RT in situ PCR.The results obtained demonstrated that Chlamydophila pneumoniae was present and alive in the tissues examined.The global concordance of results in the three techniques used was 100%. RT in situ PCR can be considered a precious tool to detect bacterial mRNA in formalin fixed paraffin embedded samples provided an optimal standardization of the key variables is achieved.

  3. 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.

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

    OpenAIRE

    Darwin Yovanny Hernández-Herrera; Andrés Mauricio Posso-Terranova; Javier Antonio Benavides; Jaime Eduardo Muñoz-Flórez; Guillermo Giovambattista; Luz Ángela Álvarez-Franco

    2011-01-01

    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 a...

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

    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.

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

    OpenAIRE

    Zidaric, V.; Janezic, S; Indra, A.; Kokotovic, Branko; Blanco, J.L.; Seyboldt, C; Diaz, C. Rodriguez; Poxton, I R; Perreten, V.; Drigo, I; Jiraskova, A; OCEPEK, M.; Weese, J.S.; Songer, J G; Rupnik, M.

    2013-01-01

    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 according 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 e...

  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. Identification of cross-contaminated animal cells by PCR and isoenzyme analysis

    OpenAIRE

    Ramya, R.; T. Nagarajan; Sivakumar, V.; Senthilkumar, R. L.; Bala Obulapathi, B.; Thiagarajan, D; Srinivasan, V.A.

    2009-01-01

    Animal cell lines have become very popular substrates for the production of vaccines and biopharmaceuticals. Characterization of candidate production cell lines is central to ensure product safety and maintenance of consistency in the manufacture of biologicals. Nested PCR and isoenzyme analysis have been used widely to prove the identity and purity of various cell lines and primary cells individually and also after deliberate cross-contamination. The nested PCR based on the Cytochrome b (Cyt...

  9. Listeria monocytogenes Identification in Food of Animal Origin Used with Real Time PCR

    OpenAIRE

    Jaroslav Pochop; Miroslava Kačániová; Lukáš Hleba; Jana Petrová; Adriana Pavelková; Ľubomír Lopašovský

    2013-01-01

    The aim of this study was to follow the contamination of food with Listeria monocytogenes by using Step One real time polymerase chain reaction (RT 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 20 samples of food of animal origin with incubation were detected strains of Listeria monocytogenes in 9 samples (swabs). Eleven samples were negative. Our results indicated that the real-time PCR ass...

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

    OpenAIRE

    Jaroslav Pochop; Miroslava Kačániová; Lukáš Hleba; Jana Petrová; Ľubomír Lopašovský; Adriana Pavelková; Alica Bobková

    2013-01-01

    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 assa...

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

    OpenAIRE

    Maria Cristina Medici; Monica Martinelli; Annalisa Aloisi; Laura Anna Abelli; Giuseppe Dettori; Carlo Chezzi

    2007-01-01

    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-...

  12. Detection of Dientamoeba fragilis in animal faeces using species specific real time PCR assay.

    Science.gov (United States)

    Chan, Douglas; Barratt, Joel; Roberts, Tamalee; Phillips, Owen; Šlapeta, Jan; Ryan, Una; Marriott, Deborah; Harkness, John; Ellis, John; Stark, Damien

    2016-08-30

    Dientamoeba fragilis is a potentially pathogenic, enteric, protozoan parasite with a worldwide distribution. While clinical case reports and prevalence studies appear regularly in the scientific literature, little attention has been paid to this parasite's biology, life cycle, host range, and possible transmission routes. Overall, these aspects of Dientamoeba biology remain poorly understood at best. In this study, a total of 420 animal samples, collected from Australia, were surveyed for the presence of Dientamoeba fragilis using PCR. Several PCR assays were evaluated for sensitivity and specificity. Two previously published PCR methods demonstrated cross reactivity with other trichomonads commonly found in animal samples. Only one assay exhibited excellent specificity. Using this assay D. fragilis was detected from one dog and one cat sample. This is the first report of D. fragilis from these animals and highlights the role companion animals may play in D. fragilis transmission. This study demonstrated that some published D. fragilis molecular assays cross react with other closely related trichomonads and consequently are not suitable for animal prevalence studies. PMID:27523936

  13. 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.

  14. Comparative identification of Candida species isolated from animals using phenotypic and PCR-RFLP methods

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    Nadăş George Cosmin

    2014-06-01

    Full Text Available The aim of this study was to identify 58 Candida sp. strains isolated from animals using the Chromatic Candida test, the API 20 C AUX system, and polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP. The Chromatic Candida test was able to identify only C. albicans and C. krusei. The API 20 C AUX system and PCR-RFLP had similar specificity for the identification of Candida strains. In case of both methods, Candida albicans was the most frequently isolated species - 22 (37.93% strains, followed by Candida krusei - 17 (29.31% strains, Candida famata - 10 (17.24% strains, Candida parapsilosis - five (8.62% strains, and Candida kefyr - four (6.89% strains. PCR-RFLP represents a reliable, quick and relatively inexpensive genotyping method, recommended for rapid identification of Candida spp.

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

    Science.gov (United States)

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

    2015-11-01

    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 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. PMID:26571269

  16. Listeria monocytogenes Identification in Food of Animal Origin Used with Real Time PCR

    Directory of Open Access Journals (Sweden)

    Jaroslav Pochop

    2013-10-01

    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 (RT 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 20 samples of food of animal origin with incubation were detected strains of Listeria monocytogenes in 9 samples (swabs. Eleven 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. Nested RT-PCR for ante mortem diagnosis of rabies from body secretion/excretion of animals suspected for rabies

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    M Dandale

    Full Text Available Aim: The present study deals with molecular technique Nested RT-PCR for detection of rabies viral RNA from biological fluid samples (Saliva, Milk and Urine collected from animal suspected for rabies and to compare the sensitivity of Nested RT-PCR applied for ante mortem diagnosis of rabies with conventional technique (immunofluorescence applied on neural tissue. Materials and Methods: Nested RT-PCR was applied on 62 biological fluid specimens collected from rabies suspected animals. First round amplification with nested set of primers (RabN1 and RabN5 yielded 1477 bp product while amplification with second round primers (RabNfor and RabNrev yielded 762 bp product. Sensitivity of the technique was compared in accordance with WHO recommended gold standard test viz. Immunofluorescence (FAT applied on brain samples. Results: By Nested RT-PCR, viral RNA could be detected in 9/24 (37.50% saliva samples, 2/17 (11.76% milk samples and 6/21 (28.57% urine samples. Confirmatory diagnosis by Immunofluorescence performed on brain sample revealed 18 true positive cases. Overall, Sensitivity of Nested RT-PCR technique employed on fluid samples was 69.23% when compared with immunofluorescence performed on brain samples. Conclusions: Early reliable ante mortem diagnosis of rabies can be obtained from biological fluid samples of animals suspected to be rabid when tested with Nested RT-PCR technique. [Vet World 2012; 5(11.000: 690-693

  3. Use of Repetitive DNA Sequences and the PCR To Differentiate Escherichia coli Isolates from Human and Animal Sources

    OpenAIRE

    Dombek, Priscilla E.; Johnson, LeeAnn K.; Zimmerley, Sara T.; Michael J Sadowsky

    2000-01-01

    The rep-PCR DNA fingerprint technique, which uses repetitive intergenic DNA sequences, was investigated as a way to differentiate between human and animal sources of fecal pollution. BOX and REP primers were used to generate DNA fingerprints from Escherichia coli strains isolated from human and animal sources (geese, ducks, cows, pigs, chickens, and sheep). Our initial studies revealed that the DNA fingerprints obtained with the BOX primer were more effective for grouping E. coli strains than...

  4. Microfluidic high-throughput reverse-transcription quantitative PCR analysis of liver gene expression in lactating animals

    International Nuclear Information System (INIS)

    We have evaluated a microfluidic lab-on-chip quantitative reverse transcription (RT) quantitative PCR (qPCR) method by measuring the expression of key actors of liver metabolism in lactating cattle. Animals in the early and in the late lactation phases were chosen because of the extreme adaptations in gene expression expected to occur. During the lactation cycle, 28 out of 48 genes were significantly regulated, notably in the same direction as previously shown by other techniques. This demonstrates that this high-throughput platform represents an attractive alternative to microarrays due to its ease of application, rapidity and lower costs. A set of 13 genes was identified—in combination with a dynamic PCA algorithm—that allowed the clearest separation between the two physiologically different groups. This paves the way for classification and diagnosis of animals in different metabolic situations by a reliable microfluidic RT-qPCR assay. (author)

  5. 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

  6. 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

  7. Direct and rapid detection by PCR of Erysipelothrix sp. DNAs prepared from bacterial strains and animal tissues.

    Science.gov (United States)

    Takeshi, K; Makino, S; Ikeda, T; Takada, N; Nakashiro, A; Nakanishi, K; Oguma, K; Katoh, Y; Sunagawa, H; Ohyama, T

    1999-12-01

    A PCR method for rapid screening of Erysipelothrix spp. in the slaughterhouse was carried out by using four species-specific sets of oligonucleotide primers after initial amplification with the primer set MO101-MO102, which amplifies the 16S rRNA sequences of all four Erysipelothrix species. The DNA sequences coding for the rRNA gene cluster, including 16S rRNA, 23S rRNA, and the noncoding region downstream of 5S rRNA, were determined in order to design primers for the species-specific PCR detection system. The homology among the 4.5-kb DNA sequences of the rRNA genes of Erysipelothrix rhusiopathiae serovar 2 (DNA Data Bank of Japan accession no. AB019247), E. tonsillarum serovar 7 (accession no. AB019248), E. rhusiopathiae serovar 13 (accession no. AB019249), and E. rhusiopathiae serovar 18 (accession no. AB019250) ranged from 96.0 to 98.4%. The PCR amplifications were specific and were able to distinguish the DNAs from each of the four Erysipelothrix species. The results of PCR tests performed directly with tissue specimens from diseased animals were compared with the results of cultivation tests, and the PCR tests were completed within 5 h. The test with this species-specific system based on PCR amplification with the DNA sequences coding for the rRNA gene cluster was an accurate, easy-to-read screening method for rapid diagnosis of Erysipelothrix sp. infection in the slaughterhouse. PMID:10565937

  8. Direct and Rapid Detection by PCR of Erysipelothrix sp. DNAs Prepared from Bacterial Strains and Animal Tissues

    Science.gov (United States)

    Takeshi, Kouichi; Makino, Souichi; Ikeda, Tetsuya; Takada, Noriko; Nakashiro, Atsushi; Nakanishi, Kazunori; Oguma, Keiji; Katoh, Yoshinobu; Sunagawa, Hiroyuki; Ohyama, Tohru

    1999-01-01

    A PCR method for rapid screening of Erysipelothrix spp. in the slaughterhouse was carried out by using four species-specific sets of oligonucleotide primers after initial amplification with the primer set MO101-MO102, which amplifies the 16S rRNA sequences of all four Erysipelothrix species. The DNA sequences coding for the rRNA gene cluster, including 16S rRNA, 23S rRNA, and the noncoding region downstream of 5S rRNA, were determined in order to design primers for the species-specific PCR detection system. The homology among the 4.5-kb DNA sequences of the rRNA genes of Erysipelothrix rhusiopathiae serovar 2 (DNA Data Bank of Japan accession no. AB019247), E. tonsillarum serovar 7 (accession no. AB019248), E. rhusiopathiae serovar 13 (accession no. AB019249), and E. rhusiopathiae serovar 18 (accession no. AB019250) ranged from 96.0 to 98.4%. The PCR amplifications were specific and were able to distinguish the DNAs from each of the four Erysipelothrix species. The results of PCR tests performed directly with tissue specimens from diseased animals were compared with the results of cultivation tests, and the PCR tests were completed within 5 h. The test with this species-specific system based on PCR amplification with the DNA sequences coding for the rRNA gene cluster was an accurate, easy-to-read screening method for rapid diagnosis of Erysipelothrix sp. infection in the slaughterhouse. PMID:10565937

  9. 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

  10. 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...

  11. Comparison of PCR with blood smear and inoculation of small animals for diagnosis of Babesia microti parasitemia.

    OpenAIRE

    Krause, P J; Telford, S; Spielman, A.; Ryan, R; Magera, J; Rajan, T V; Christianson, D; Alberghini, T V; Bow, L; Persing, D

    1996-01-01

    The specific diagnosis of babesiosis, which is caused by the piroplasm Babesia microti, is made by microscopic identification of the organism in Giemsa-stained thin blood smears, detection of babesial antibody in acute-and convalescent-phase sera, or identification of the organism following the injection of patient blood into laboratory animals. Although rapid diagnosis can be made with thin blood smears, parasites are often not visualized early in the course of infection. PCR is a new, rapid...

  12. 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.

  13. Evaluación del bienestar animal mediante indicadores conductuales en granjas pequeñas de ovinos.

    OpenAIRE

    Otero Prevost, Luis Gabriel

    2013-01-01

    El bienestar animal hace mucho que dejó de ser una postura filosófica contra los sistemas de manejo intensivo. Ahora visto como garantía de alta calidad en procesos pecuarios, los consumidores de todo el mundo comienzan a exigir productos animales en condiciones que eviten el maltrato y sufrimiento. Tal situación hace necesaria la aplicación de protocolos y criterios de evaluación que basados en metodologías integrales, mejoren el manejo, instalaciones, sanidad e higiene en las diferentes es...

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

    OpenAIRE

    A. Pérez Rico; J. Lacasa Navarro; J.M. Rubio Muñoz; S. Ruiz Contreras; J.L. Vega Pla

    2013-01-01

    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...

  15. 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 according...

  16. STUDY OF PERSISTENT VIRAL INFECTION IN AN ANIMAL MODEL OF VIRAL MYOCARDITIS BY PCR

    Institute of Scientific and Technical Information of China (English)

    马睿; 陈曙霞; 刘晶星

    2000-01-01

    ffeStnn6 Objectif Etudier ie r6le de l'infection virale persistante dans ie pethog4de de la myOCardite virale.ANt~ L' ARN viral dens ie my~rde et ie mug et l' alteration potholedque du m~rde ent ate ewilnd per la techniquede PCR adns un mangle de myrmrdite virale chez ies ~ris. Rhaltats L 'ARN viral a ate detects an 3'jour dens ie mug etie myrmrde. An 8'jour, I 'ARN viral an niveau du mug a ate pertiellement dewnu then f lorsque l' alteration pethologiquedu myocarde a atteint un maximum. he 12'jour, L' ARN ...

  17. 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

  18. Quantitative PCR measurements of Escherichia coli including shiga toxin-producing E. coli (STEC) in animal feces and environmental waters.

    Science.gov (United States)

    Ahmed, W; Gyawali, P; Toze, S

    2015-03-01

    Quantitative PCR (qPCR) assays were used to determine the concentrations of E. coli including shiga toxin-producing E. coli (STEC) associated virulence genes (eaeA, stx1, stx2, and hlyA) in ten animal species (fecal sources) and environmental water samples in Southeast Queensland, Australia. The mean Log10 concentrations and standard deviations of E. coli 23S rRNA across fecal sources ranged from 1.3 ± 0.1 (horse) to 6.3 ± 0.4 (cattle wastewater) gene copies at a test concentration of 10 ng of DNA. The differences in mean concentrations of E. coli 23S rRNA gene copies among fecal source samples were significantly different from each other (P cattle wastewater samples ranged from 3.8 to 5.0 gene copies at a test concentration of 10 ng of DNA. Of the 18 environmental water samples tested, three (17%) were positive for eaeA and two (11%) samples were also positive for the stx2 virulence genes. The data presented in this study will aid in the estimation of quantitative microbial risk assessment (QMRA) from fecal pollution of domestic and wild animals in drinking/recreational water catchments. PMID:25648758

  19. Comparison of ELISA and PCR vis-à-vis cultural methods for detecting Aeromonas spp. in foods of animal origin.

    Science.gov (United States)

    Arora, S; Agarwal, R K; Bist, B

    2006-02-01

    The present study was conducted to assess the best method of the most commonly used methods for detection of aeromonads in foods of animal origin. With this objective an OMP based indirect plate ELISA and a duplex-PCR using primers targeting aerolysin gene and 16S rRNA gene and yielding amplicons of 252 bp and 599 bp, respectively, were standardized. The standardized protocols and the conventional cultural method were then compared for their respective sensitivities and specificities for detecting aeromonads from chicken and milk samples. Both the standardized assays were found to be highly specific for Aeromonas. The efficiency of the standardized indirect-ELISA and duplex-PCR protocols was assessed by artificial inoculation studies with varying concentrations of Aeromonas cells inoculated in chicken and milk samples followed by enrichment in Alkaline Peptone Water supplemented with 10 mg/ml cephalothin (APW-C) for 12 h. The results revealed that indirect-ELISA was able to detect a minimum of 10(3) cells/ml or g of Aeromonas cells in spiked milk and chicken samples, respectively. Whereas, duplex-PCR and cultural method were able to detect as low as 1 cell/ml or g of Aeromonas cells in spiked milk and chicken samples. The developed assays were also tested for their efficiency to detect Aeromonas spp. in naturally contaminated milk and chicken samples. Out of a total 50 milk samples screened for presence of Aeromonas by the three methods viz., indirect-ELISA, duplex-PCR and cultural method only 1 (2%) turned out to be positive showing positive results by all three methods. Similarly, 50 samples of chicken were tested by all three methods. Three samples (6%) turned out to be positive and here again by all the three methods. PMID:16216375

  20. 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 n...

  1. 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.

  2. Análisis de la expresión transcripcional del receptor de estrógeno en ovario de ovejas prepúberes de razas Texel y Criolla Araucana mediante RT-PCR cuantitativo en tiempo real

    Directory of Open Access Journals (Sweden)

    M Flores

    2015-01-01

    Full Text Available El estado morfofuncional del sistema reproductivo de las ovejas es determinado por las hormonas sexuales, que actúan por medio de receptores específicos, desencadenando una serie de cambios celulares, metabólicos y proliferativos dependientes de la expresión de numerosos genes. A diferencia de otros mamíferos, las ovejas presentan en el endometrio y posiblemente otros órganos del sistema reproductivo receptores de estrógenos fisiológicamente activos desde la etapa prepuberal cuya función aún no está esclarecida. La información sobre la expresión de receptores de hormonas sexuales en el aparato reproductor es muy escasa, sobre todo en el ovario y no existen estudios que correlacionen la raza con la expresión de estos receptores. Generalmente los criadores privilegian razas de mayor nivel de prolificidad y esto podría estar relacionado con el nivel de expresión de los receptores de estrógeno en el sistema reproductivo. El objetivo del presente trabajo fue evaluar comparativamente la expresión transcripcional del receptor de estrógeno en el ovario de ovejas prepúberes de raza Texel de alta prolificidad y de raza Criolla Araucana de prolificidad estándar, mediante análisis de RT-PCR en tiempo real cuantitativo.

  3. 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. PMID:26827864

  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

    in highly heterogeneous species, sequence divergence was observed and more than one PCR-DGGE profile was obtained. Application of the PCR-DGGE method to DNA extracted from faeces of zoo animals revealed the presence of Helicobacter DNA in 13 of 16 samples; a correlation was seen between the mobility of PCR...... on purified amplicons. Sixteen DGGE profiles were derived from 44 type and reference strains of 20 Helicobacter species, indicating the potential of this approach for resolving infection of a single host by multiple Helicobacter species. Some more highly related species were not differentiated whereas...... products in DGGE analysis and DNA sequencing. In combination, this indicated that zoo animals are colonized by a wide range of different Helicobacter species; seven animals appeared to be colonized by multiple Helicobacter species. By this approach, presumptive identifications were made of Helicobacter...

  5. 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...

  6. 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

  7. 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

  8. 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.

  9. Diagnóstico y tipificación del virus de la leucosis bovina mediante una prueba de PCR-RFLP a partir de ADN extraído desde células somáticas de la leche Diagnosis and typing of bovine leukaemia virus using a PCR-RFLP test on DNA extracted from somatic cells in milk

    Directory of Open Access Journals (Sweden)

    R Felmer

    2006-01-01

    Full Text Available Se evaluó la factibilidad de aplicar una prueba de PCR para el diagnóstico y tipificación molecular del virus de la leucosis enzoótica bovina (VLB, directamente desde muestras de leche. Se analizó un total de 40 muestras de estanque predial, 33 de las cuales fueron seropositivas a una prueba de ELISA. El PCR confirmó la presencia del virus en el 100% de los estanques seropositivos, mientras que en las muestras seronegativas no se obtuvo una banda de amplificación. El posterior análisis de estas muestras mediante RFLP permitió identificar la presencia de 2 de los 3 subgrupos conocidos de variantes genéticas del virus. La aplicación de esta prueba en 10 animales de un predio permitió confirmar la presencia de más de una variante genética dentro del mismo predio, sugiriendo la probable reinfección de este predio con otras cepas del virus. Es necesario destacar que este trabajo constituye el primer reporte de la aplicación de una prueba de PCR-RFLP para la detección y tipificación del VLB directamente desde muestras de leche. De esta forma, la técnica de PCR descrita se puede utilizar no sólo como complemento al diagnóstico del VLB, sino también como una forma conveniente de realizar la tipificación del virus en una zona determinada o incluso a nivel de país, lo cual proporciona una forma rápida y conveniente de estudiar la epidemiología y distribución de la infección del VLB en nuestros rebaños.The aim of this study was to assess the suitability of using DNA isolated from milk somatic cells for the diagnosis and molecular typing of bovine leukaemia virus (BLV. A total of 40 bulk milk samples were analysed and thirty three of them resulted seropositive to BLV after being evaluated using an indirect ELISA test. A PCR test confirmed the presence of the virus in all 33 seropositive samples whereas in the remaining seronegative samples it was not possible to detect a specific band for the virus. A RFLP analysis identified 2

  10. 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

  11. Animals

    International Nuclear Information System (INIS)

    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)

  12. The development of a hexaplex-conventional PCR for identification of six animal and plant species in foodstuffs.

    Science.gov (United States)

    Safdar, Muhammad; Junejo, Yasmeen

    2016-02-01

    A hexaplex-conventional PCR assay was developed for identification of five meat and one plant species origins in foodstuffs simultaneously. The method merges the use of horse (Equus caballus), soybean (Glycine max), sheep (Ovis aries), poultry (Meleagris meleagris), pork (Sus scrofa), and cow (Bos taurus) specific primers that amplify fragments (horse; 85 bp, soybean; 100 bp, sheep; 119 bp, poultry; 183 bp, pork; 212 bp and cow; 271 bp) of the mitochondrial cyt b, lectin, 12S rRNA, 12S rRNA, ATPase subunit 6 genes and ATPase subunit 8 genes respectively, and a universal 18S rRNA primers that amplifies a 141 bp. Multiplex analysis of the reference food samples showed that detection limit of the hexaplex assay was 0.01% for each species. Taken together, all data indicated that this hexaplex PCR assay was a simple, fast, sensitive, specific, and cost-effective detection method for horse, soybean, sheep, poultry, pork and cow species in foodstuffs. PMID:26304406

  13. 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.

  14. 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.

  15. 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).

  16. Comparison of immunocapture and RT-PCR techniques for the detection of peste-des-petits-ruminants virus (PPRV) in eye and nose swabs from infected animals

    International Nuclear Information System (INIS)

    Full text: Peste des Petits Ruminants (PPR) is a highly contagious disease of domestic and wild small ruminants. It constitutes a major constraint on production in areas where it is endemic. Classically, it is characterised by fever, nasal and ocular discharges, diarrhoea, respiratory distress, mucosal erosive lesions and death in 40-80% of acute cases. All these clinical signs, apart from the respiratory symptoms, are very similar to those of rinderpest (RP). The causal agents of both diseases are viruses which belong to the Morbillivirus genus. Described for the first time in 1942 in Cote d'Ivoire, PPR was considered for a long time as a disease of West African countries. The current knowledge on its epidemiology shows that this is no longer true, nor it is a solely African disease since it is widespread in countries lying between the Sahara and the Equator, in the Middle East and in South-West Asia. These data indicate that PPR existed undetected in most of the known endemic areas for a long time. It was overlooked because of the similarity of clinical signs to rinderpest as indicated above and also to pasteurellosis for the bronchopneumonia. The current knowledge of the disease has grown up quickly once specific diagnostic tests became available in the 1990's: serological diagnosis in the competitive format or for antigen detection by immunocapture cDNA probe and the amplification (RT-PCR) technique for the nucleic acid detection. While the immunocapture (ICE) can detect virus up to 100.6 TCID50 of virus in 50μl of sample, the limit of detection of the RT-PCR is estimated to 0.001 TCD50/ml. During a study to analyse the pathogenicity of some PPRV isolates, we have compared the efficiency of the ICE test, antibody-based antigen detection, and RT-PCR, with the RT-PCR, nucleic acid-based detection technique, to detect the shedding of the virus by the infected animals. For the study, Sahelian goats were inoculated IM with PPRV Guinea (isolate from Guinea Conakry

  17. Mycoplasma diagnosis by PCR from bedding of mycoplasmal dairy herds and association with disease in dairy animals

    International Nuclear Information System (INIS)

    Full text: Infection with Mycoplasma spp, typically M. bovis, is an important disease complex of dairy cattle. Mycoplasma spp can cause mastitis, arthritis, metrititis, pneumonia, septicemia, and death of cattle. Standard microbial cultures of milk samples do not isolate Mycoplasma spp; special methods are necessary. Mycoplasma infections have been reported as contagious in nature, primarily by milking machines and respiratory spread. Bulk tank milk samples (n = 5 samples per tank) were collected from all bulk tanks on most dairy farms in Utah, USA (n = 222 farms, 292 tanks) at 3-4 day intervals, resulting in a sensitivity of 97% for Mycoplasma spp. Mycoplasma was detected on 16/222 dairy farms in Utah (7%), a relatively high prevalence compared to the rest of the USA. After initial surveillance, follow up was conducted on positive farms. One farm milking approximately 4500 Holstein cows in dry lot and free stall housing experienced an outbreak of clinical mastitis (CM) caused by Mycoplasma spp., affecting 35 cows per month vs. the endemic rate of approximately 3 CM cases per month (aseptic milk samples from all CM cases were cultured from this herd). Bedding sand was used following a recycling and manure separation process on the farm; sand samples were cultured for mycoplasmas and other bacteria during the outbreak. Acholeplasma laidlawii was found in one sample, 2 samples were positive for M. bovis by PCR, and one month later 14/20 cow pens' and bedding samples tested Modified Hayflick medium culture-positive for Mycoplasma spp. (testing by 3 different laboratories). During the same month, one recycled bedding sand sample and one cow pen sand sample tested PCR-positive at the Utah Veterinary Diagnostic Laboratory; amplicon sequencing of both isolates showed 99% homology with M. bovis. Positive bedding sand (18,000 kg) was transported from the farm to Utah State University and stored in a pile outdoors. As the weather progressed from late winter (March) to summer

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

    OpenAIRE

    Avila Barrera, Erica Natalia

    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 ...

  19. 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

    OpenAIRE

    Nolberto Arismendi S; Nancy Andrade S; Ricardo Riegel Sch; Roberto Carrillo Ll.

    2010-01-01

    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 identif...

  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

    OpenAIRE

    Lopez, J.C.; R Mcfarlane; Ulloa, J.

    2006-01-01

    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 compara...

  1. Development of a High-Throughput Multiplex PCR and Capillary Electrophoresis Technique for Serotype Determination of Salmonella Enterica Food Animal Isolates

    Science.gov (United States)

    Background: Previously, a multiplex PCR technique was developed to identify the top 30 human clinical serotypes of Salmonella enterica. To improve the speed, ease of use, utility and discriminatory ability of the technique, additional primers were added and the PCR product discrimination and analysi...

  2. 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

  3. PCR survey of 50 introns in animals: cross-amplification of homologous EPIC loci in eight non-bilaterian, protostome and deuterostome phyla

    OpenAIRE

    Gérard, Karin; Guilloton, Edith; Arnaud-Haond, Sophie; Aurelle, Didier; Bastrop, Ralf; Chevaldonné, Pierre; Derycke, Sophie; Hanel, Reinhold; Lapègue, Sylvie; Lejeusne, Christopher; Mousset, Sylvain; Ramsak, Andreja; Remerie, Thomas; Viard, Frédérique; Feral, Jean-Pierre

    2013-01-01

    Exon Primed Intron Crossing (EPIC) markers provide molecular tools that are susceptible to be variable within species while remaining amplifiable by PCR using potentially universal primers. In this study we tested the possibility of obtaining PCR products from 50 EPIC markers on 23 species belonging to seven different phyla (Porifera, Cnidaria, Arthropoda, Nematoda, Mollusca, Annelida, Echinodermata) using 70 new primer pairs. A previous study had identified and tested those loci in a dozen s...

  4. 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...

  5. DETECCIÓN Y CUANTIFICACIÓN DE Spongospora subterranea f. sp. subterranea EN PLANTAS SEÑUELO Y CULTIVOS DE PAPA EN COLOMBIA MEDIANTE qPCR Detection and Quantification of Spongospora subterranea f. sp. subterranea in Bait Plants and Potato Fields in Colombia using qPCR

    Directory of Open Access Journals (Sweden)

    NEVAR GARCÍA BASTIDAS

    2013-04-01

    Full Text Available La sarna polvosa de la papa (Solanum tuberosum, S. phureja causada por Spongospora f. sp. subterranea (Sss, es una de las enfermedades más limitantes de este cultivo. En Colombia, se han empleado diferentes métodos de detección asintomática de Sss, incluyendo bioensayos con plantas señuelo, PCR de ITS y pruebas de ELISA. Sin embargo, sus niveles de sensibilidad son bajos o requieren tiempos extensos. Una alternativa para complementar dichas herramientas es la PCR cuantitativa en tiempo real (qPCR. En este trabajo se evaluó dicha técnica utilizando los juegos de cebadores SsTQF1-SsTQR1; Spon421F-Spon494R y SscolF-SscolR (diseñados en este estudio, bajo la metodología de SYBR Green®; mientras que con Taqman® se evaluaron los cebadores SponFSponR y la sonda SponP. Una vez determinada la funcionalidad de los cebadores, se descartó por inespecificidad, el par Spon421F-Spon494R ; para los restantes se realizaron curvas estándar basadas en diluciones seriadas de quistosoros. Las pruebas de qPCR detectaron a Sss en las 20 muestras evaluadas de plantas señuelo de Nicotiana benthamiana y papa, utilizando los cebadores SsTQF1-SsTQR1 (Ct: 10,57-29,34 y SscolF-SscolR (Ct: 14,39-34,08; mientras que 19 de las muestras fueron positivas con SponF-SponR-SponP (Ct: 15,63-38,93. A partir de 20 muestras de raíces de papa de cultivos de La Unión (Antioquia, Colombia, fue posible detectar el patógeno en 17 de ellas con SscolF-SscolR, estimándose una concentración de 6470 a 1,39 x 10(10 quistorosos/mL. Estos resultados indican la ocurrencia de altos niveles de inóculo de Sss en esta región y enfatizan en la necesidad de fortalecer los programas de certificación de tubérculo-semilla en Colombia.In recent years, potato crops (Solanum tuberosum, S. phureja have been seriously affected by powdery scab; a disease caused by Spongospora subterranea f.sp. subterranea (Sss. In Colombia, asymptomatic detection of Sss has been achieved with bait plants, PCR

  6. 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 ...

  7. 动物源性食品鸭血、猪血DNA提取及多重PCR鉴别研究%DNA extraction and multiple PCR distinction research of animal food duck blood and pig blood

    Institute of Scientific and Technical Information of China (English)

    吕二盼; 周正; 周巍; 李洋洋; 张薇; 吴涛; 曾小盼; 李波; 张伟

    2012-01-01

    目的:研究从鸭血、猪血中提取DNA的快速简便方法并建立多重PCR鉴别方法。方法:用KI提取法从固体块状鸭血、猪血中提取DNA,经PCR扩增检测提取效果。建立多重PCR方法鉴别动物源性食品中的鸭血、猪血成分,并对市售动物源性血制品进行检测。结果:这种方法提取到的DNA纯度较高,凝胶电泳条带整齐,背景清晰;PCR反应能扩增出目的条带。多重PCR能同时扩增出鸭和猪的条带。结论:这种改进的DNA抽提方法能获得高纯度DNA,比传统方法安全、简便、节省试剂,PCR扩增结果很好,应用多重PCR方法能同时检测出血样制品中的鸭、猪成分。%Objective Study on duck blood and pig blood aimed to find a quick and easy way to extract DNA and establish multiple PCR method for identification. Methods=The KI method was used for DNA extracting from the solid massive duck blood and pig blood and was detected by PCR amplification. A multiplex PCR method was established to identify the duck,pig blood ingredients in the food of animal original and carried on the examination to animal blood products from the market. Results=The gel electrophoresis stripes indicated that this DNA extraction method was of high purity,clear and neat. And the target bands could be amplified by PCR. Multiplex PCR simultaneously amplified duck and pig bands. Conclusion=The improved DNA extraction methods could obtain the DNA of high purity. It was safe,convenient and economical compared with the traditional method. PCR. Amplification result was good,the application of multiplex PCR method could also detect duck and pig ingredients in the blood product sample.

  8. 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

  9. DETECCIÓN Y DIFERENCIACIÓN DE Mycoplasma gallisepticum Y Mycoplasma synoviae MEDIANTE LA TÉCNICA DE PCR A PARTIR DE HISOPOS TRAQUEALES DE AVES CON SÍNTOMAS RESPIRATORIOS Detection and Differentiation of Mycoplasma gallisepticum and Mycoplasma synoviaeby PCR from Tracheal Swabs from Birds with Respiratory Symptoms

    Directory of Open Access Journals (Sweden)

    CESAR E VENTURA

    2012-12-01

    Full Text Available Los micoplasmas son importantes patógenos en las aves por ser responsables de cuadros respiratorios que ocasionan grandes pérdidas económicas a la industria avícola alrededor del mundo. Existen principalmente dos especies de micoplasmas como causantes de enfermedad en aves comerciales, el Mycoplasma gallisepticum(MG y el Mycoplasma synoviae(MS. Teniendo en cuenta su importancia y la necesidad de conocer y diferenciar las diferentes especies de micoplasmas presentes en las explotaciones avícolas, se tomaron 91 muestras de hisopos traqueales de aves con síntomas respiratorios, provenientes de igual número de granjas de pollo de engorde, ponedoras comerciales y reproductoras pesadas ubicadas en los departamentos de Cundinamarca y Boyacá, Colombia, y se determinó la presencia de MG y MS por la técnica de PCR. La prevalencia determinada fue de 39,6 % para MG y 47,3 % para MS, encontrándose diferencias estadísticamente significativas cuando se comparó la positividad a MG y MS y el tipo de explotación (p Mycoplasmas are worldwide pathogens that affect the poultry industry causing respiratory illness which cause a negative economic impact. Two mycoplasmas species are the most important in the commercial poultry: Mycoplasma gallisepticum(MG and Mycoplasma synoviae(MS. By its importance and necessity to know and differentiate between mycoplasmas species in locals poultry houses this study used the PCR technique like a diagnosis tool, using tracheal swabs from bird with respiratory symptoms. A total of 91 samples from broilers, layers and breeders farms located in the departments of Cundinamarca and Boyacá was processed. The punctual prevalence founded in this study was 39.6 % for MG and 47.3 % for MS. Statistical differences for type of production and positive samples for MG y MS (p < 0.05 were founded, a bigger number of positive samples from layers and breeder in comparison to broilers was found. In the same way, the positive samples for

  10. 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.

  11. Molecular diagnostic PCR handbook

    International Nuclear Information System (INIS)

    The uses of nucleic acid-directed methods have increased significantly in the past five years and have made important contributions to disease control country programmes for improving national and international trade. These developments include the more routine use of PCR as a diagnostic tool in veterinary diagnostic laboratories. However, there are many problems associated with the transfer and particularly, the application of this technology. These include lack of consideration of: the establishment of quality-assured procedures, the required set-up of the laboratory and the proper training of staff. This can lead to a situation where results are not assured. This book gives a comprehensive account of the practical aspects of PCR and strong consideration is given to ensure its optimal use in a laboratory environment. This includes the setting-up of a PCR laboratory; Good Laboratory Practice and standardised PCR protocols to detect animal disease pathogens. Examples of Standard Operating Procedures as used in individual specialist laboratories and an outline of training materials necessary for PCR technology transfer are presented. The difficulties, advantages and disadvantages in PCR applications are explained and placed in context with other test systems. Emphasis is placed on the use of PCR for detection of pathogens, with a particular focus on diagnosticians and scientists from the developing world. It is hoped that this book will enable readers from various disciplines and levels of expertise to better judge the merits of PCR and to increase their skills and knowledge in order to assist in a more logical, efficient and assured use of this technology

  12. PCR survey of 50 introns in animals: cross-amplification of homologous EPIC loci in eight non-bilaterian, protostome and deuterostome phyla.

    Science.gov (United States)

    Gérard, K; Guilloton, E; Arnaud-Haond, S; Aurelle, D; Bastrop, R; Chevaldonné, P; Derycke, S; Hanel, R; Lapègue, S; Lejeusne, C; Mousset, S; Ramšak, A; Remerie, T; Viard, F; Féral, J-P; Chenuil, A

    2013-12-01

    Exon Primed Intron Crossing (EPIC) markers provide molecular tools that are susceptible to be variable within species while remaining amplifiable by PCR using potentially universal primers. In this study we tested the possibility of obtaining PCR products from 50 EPIC markers on 23 species belonging to seven different phyla (Porifera, Cnidaria, Arthropoda, Nematoda, Mollusca, Annelida, Echinodermata) using 70 new primer pairs. A previous study had identified and tested those loci in a dozen species, including another phylum, Urochordata (Chenuil et al., 2010). Results were contrasted among species. The best results were achieved with the oyster (Mollusca) where 28 loci provided amplicons susceptible to contain an intron according to their size. This was however not the case with the other mollusk Crepidula fornicata, which seems to have undergone a reduction in intron number or intron size. In the Porifera, 13 loci appeared susceptible to contain an intron, a surprisingly high number for this phylum considering its phylogenetic distance with genomic data used to design the primers. For two cnidarian species, numerous loci (24) were obtained. Ecdysozoan phyla (arthropods and nematodes) proved less successful than others as expected considering reports of their rapid rate of genome evolution and the worst results were obtained for several arthropods. Some general patterns among phyla arose, and we discuss how the results of this EPIC survey may give new insights into genome evolution of the study species. This work confirms that this set of EPIC loci provides an easy-to-use toolbox to identify genetic markers potentially useful for population genetics, phylogeography or phylogenetic studies for a large panel of metazoan species. We then argue that obtaining diploid sequence genotypes for these loci became simple and affordable owing to Next-Generation Sequencing development. Species surveyed in this study belong to several genera (Acanthaster, Alvinocaris, Aplysina

  13. 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. PMID:25245143

  14. 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 .

  15. Method for detecting, identifying and quantifying peronospora arborescens by real time quantitative PCR

    OpenAIRE

    Landa, Blanca B.; Jiménez-Díaz, Rafael M.; Montes Borrego, Miguel

    2009-01-01

    [ES] El método para la cuantificación de peronospora arborescens por PCR cuantitativa (qPCR) en una muestra biológica comprende extraer el ADN contenido en dicha muestra biológica y amplificarlo mediante qPCR. De aplicación en la cuantificación de P. arborescens

  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. Clasificación en subtipos moleculares de tumores de mama de pequeños animales mediante métodos inmunohistoquímicos Classification in molecular subtypes of breast tumors of small animals through immunohistochemical methods

    Directory of Open Access Journals (Sweden)

    Mª V. Ortega García

    2013-03-01

    methods in mammary tumors of small animals to classify them in molecular subtypes and their association with the invasion, grade and histological type of the malignancies. Material and Methods: samples of malignant mammary tumors, 10 from canine species and 3 from feline ones. Internal positive control: non-tumoral mammary gland adjacent to the malignancy. Results: 23% (3/13 of the tumors were of the luminal B subtype, 23% (3/13 were HER2 positive, 46% (6/13 were basal types and 7,6% (1/13 were unclassifiable because they did not express any of the tested tumor markers. None of the cases belonged to the luminal A subtype. The 6 basal tumors were grade II or III and presented only stromal infiltration or vascular invasion as well. Two thirds of the HER2 positive tumors presented stromal infiltration and half the tumors were grade II. Two thirds of luminal B tumors were grade II or III. All internal controls were positive. There were no significant differences in the distribution of the molecular subtypes among the different groups of the invasion (p-value=0.26 and malignancy grade variables (p-value=0.42. There were differences of borderline statistical significance in the distribution of the molecular subtypes among the different groups of the histological type variable (p-value=0.08. Conclusions: the application of the antibodies panel has allowed to find 4 (luminal B, HER2, basal and unclassified out of 5 possible molecular subtypes.

  18. Evaluación diagnóstica de fracciones cromatográficas de Fasciola hepatica mediante Western Blot y ELISA en animales infectados Diagnostic evaluation of chromatographic fractions of Fasciola hepatica by Western Blot and ELISA in infected animals

    Directory of Open Access Journals (Sweden)

    F. FREDES

    1997-01-01

    Full Text Available La fasciolosis causada por Fasciola hepatica se diagnostica rutinariamente mediante el examen coprológico. Debido a que este examen no es 100% sensible y además es ineficaz en la etapa pre-patente, se realizó este trabajo con el objeto de caracterizar y seleccionar fracciones antigénicas de valor diagnóstico de extractos de excreción-secreción del parásito. Se utilizó cromatografía de exclusión por tamaño molecular (Sephacryl S-300, electroforesis en geles de poliacrilamida en ambiente reductor (SDS-PAGE y posterior western blot (WB, además de un método de "enzyme-linked immuno-sor-bent assay" (ELISA en microplaca. Para evaluar el valor diagnóstico de los antígenos se usaron sueros de las especies ovina, porcina y equina en tres estados (sanos, con otras parasitosis y naturalmente infectados con F. hepatica. Mediante el método cromatográfico se obtuvieron hasta 5 "peaks", que interpolados en una curva patrón representaron polipéptidos de pesos aproximados de 2.000, 400, 150, 29 y menores a 29 kDa. De éstos, los inmunorreactivos específicos para la enfermedad en las tres especies animales, bajo los criterios de SDS-PAGE y posterior WB, fueron los de 400, 150, 29 y The antigenic components of excretory-secretory products of adult F. hepatica, were separated by gel filtration chromatography (Sephacryl S-300 and then analized by polyacrylamide gel electrophoresis (SDS-PAGE, followed by Western Blot (WB. In order to evaluate the sensitivity, specificity and predictive value of the selected fractions an enzyme-linked immunosorbent assay (ELISA was used with sera from sheep, swine and horses infected with F. hepatica, as well as with control sera (uninfected animals. The chromatographic curve presented up to 5 peaks, representing polypeptides with a molecular weight of 2000, 400, 150, 29 and less than 29 kDa, according to the interpolation with a standard curve of commercial polypeptide molecular weights. The results obtained with

  19. Publicidad expandida mediante realidad aumentada

    OpenAIRE

    Martí Parreño, José

    2011-01-01

    La realidad aumentada, aplicada al marketing, abre numerosas oportunidades para propiciar la decisión de compra del consumidor. Éste puede ver cómo un producto “cobra vida” mediante una pantalla en la que se superponen imágenes e información digital a la del entorno real que está viendo en ese mismo momento. En España, los consumidores ya han podido ver cómo empresas como El Corte Inglés o Doritos empleaban esta novedosa tecnología para promocionar sus productos. El autor plantea, por tanto, ...

  20. Effect of Cage-Wash Temperature on the Removal of Infectious Agents from Caging and the Detection of Infectious Agents on the Filters of Animal Bedding-Disposal Cabinets by PCR Analysis.

    Science.gov (United States)

    Compton, Susan R; Macy, James D

    2015-11-01

    Efficient, effective cage decontamination and the detection of infection are important to sustainable biosecurity within animal facilities. This study compared the efficacy of cage washing at 110 and 180 °F on preventing pathogen transmission. Soiled cages from mice infected with mouse parvovirus (MPV) and mouse hepatitis virus (MHV) were washed at 110 or 180 °F or were not washed. Sentinels from washed cages did not seroconvert to either virus, whereas sentinels in unwashed cages seroconverted to both agents. Soiled cages from mice harboring MPV, Helicobacter spp., Mycoplasma pulmonis, Syphacia obvelata, and Myocoptes musculinus were washed at 110 or 180 °F or were not washed. Sentinels from washed cages remained pathogen-free, whereas most sentinels in unwashed cages became infected with MPV and S. obvelata. Therefore washing at 110 or 180 °F is sufficient to decontaminate caging and prevent pathogen transmission. We then assessed whether PCR analysis of debris from the bedding disposal cabinet detected pathogens at the facility level. Samples were collected from the prefilter before and after the disposal of bedding from cages housing mice infected with both MPV and MHV. All samples collected before bedding disposal were negative for parvovirus and MHV, and all samples collected afterward were positive for these agents. Furthermore, all samples obtained from the prefilter before the disposal of bedding from multiply infected mice were pathogen-negative, and all those collected afterward were positive for parvovirus, M. pulmonis, S. obvelata, and Myocoptes musculinus. Therefore the debris on the prefilter of bedding-disposal cabinets is useful for pathogen screening. PMID:26632784

  1. 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...

  2. DETECCIÓN DE UN DEFECTO GENÉTICO EN BOVINOS MEDIANTE UNA PRUEBA DE ADN. Detection of a bovine genetic defect by a DNA probe

    Directory of Open Access Journals (Sweden)

    Ricardo Felmer D.

    2001-01-01

    Full Text Available La deficiencia en la capacidad de unión de leucocitos bovinos a los antígenos, más conocida como BLAD, es una enfermedad hereditaria que resulta letal para el ganado de la raza Holstein. Su principal característica es ser una enfermedad autosómica recesiva que puede ser transmitida a la descendencia. El objetivo del trabajo fue estandarizar la metodología para realizar un diagnóstico de la enfermedad mediante técnicas moleculares y lograr una primera aproximación sobre la frecuencia génica del alelo mutado en algunas poblaciones de toros de la IX y X Región. En base a la amplificación del ADN mediante reacción en cadena de polimerasa (PCR y posterior digestión con enzimas de restricción Taq I y Hae III, fue posible visualizar, mediante electroforesis en gel de agarosa al 4%, los fragmentos de restricción característicos para bovinos normales, portadores o enfermos. La técnica se validó mediante un muestreo de 59 bovinos. De los 55 toros analizados uno resultó ser portador de BLAD, lo que implica una frecuencia génica de 1,79% en la población de toros de la IX y X Región. La técnica de PCR acoplada a la digestión con enzimas de restricción (Hae III y Taq I ha demostrado que identifica inequívocamente el genotipo del ganado en un locus determinado y permite el diagnóstico certero y precoz de aquellos animales enfermos y/o portadores del defecto hereditario conocido como BLAD.The adhesion deficiency of bovine leukocytes to antigens, known as BLAD (bovine leukocyte adhesion deficiency, is a hereditary genetic disease which is lethal for the Holstein breed. It is a recessive autosomal disease that can be transmitted to the offspring. The objective of this research was to standardize the molecular techniques to diagnose BLAD and also to get a first approximation of the genetic frequency of the mutated allele in the bull population of the IXth and Xth Regions. DNA amplification using polymerase chain reaction (PCR and

  3. 动物源性食品鸭血、猪血DNA提取方法研究及双重PCR检测%Study on DNA Extraction of Animal Origin Food Duck Blood and Pig Blood and Detection by Double PCR

    Institute of Scientific and Technical Information of China (English)

    周正; 吕二盼; 周巍; 张薇; 邢文静; 柳毅

    2012-01-01

    研究从鸭血、猪血中提取DNA的快速简便方法并建立双重PCR鉴别方法。用改进的氯仿-醋酸钠(NaAc)提取法和KI提取法从固体块状鸭血中提取DNA,与经典酚仿抽提法进行对比,经PCR扩增检测提取效果。建立双重PCR方法鉴别动物源性食品中的鸭血、猪血成分,并对市售动物源性血制品进行检测。这两种改进的DNA提取方法得到的DNA纯度较高,凝胶电泳条带整齐,背景清晰;PCR反应能扩增出目的条带。双重PCR能同时扩增出鸭和猪的条带。这两种改进的DNA抽提方法能获得高纯度DNA,比传统方法安全、简便、节省试剂,PCR扩增结果很好,应用双重PCR方法能同时检测出血样制品中的鸭、猪成分。%Study onduck-blood and pig blood-a-imed to-find a-quickand eas; way-io extraci-DNA-andestablish PCR method for identification. The improved Chloroform-NaAc method and KI extraction method used for DNA extracting were compared with the classic Phenol extraction method by PCR amplification from the solid massive duck blood. A double PCR method was established to identify the duck, pig blood components in animal origin food and carried on the examination to animal blood products from the market. The gel electrophoresis stripes indicated that the two improved DNA extraction methods were of high purity; the target bands were clear and neat and can be amplified by PCR. Double PCR simultaneously amplified duck and pig bands. The two improved DNA extraction methods can obtain the DNA of high purity, they are safe, convenient and economical compared with the traditional method. PCR amplification result is good, the application of duplex PCR method can also detect duck and pig components in the blood product.

  4. External PCR, ASN's decision

    International Nuclear Information System (INIS)

    The French law imposes in some situations the presence of a person skilled in radiation protection (PCR). This article describes the cases when this person must belong to the staff of the enterprise or when this person may be sub-contracted. For instance in most nuclear facilities the PCR must be on the payroll, for enterprises dedicated to nuclear transport the PCR's job can be sub-contracted. A decision given by the ASN (French Nuclear Safety Authority) sets the minimal requests (in terms of training, job contract, activities) of the sub-contracted PCR. (A.C.)

  5. 动物源性食品中志贺氏菌实时荧光定量PCR快速检测方法的建立%Development of a dual real-time PCR for the rapid detection of Shigella 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; 李一经

    2014-01-01

    根据志贺氏菌属高度保守的ipaH基因序列,设计探针和引物,通过优化反应条件,建立检测动物源性食品中志贺氏菌实时荧光定量PCR方法,应用于动物源性食品中志贺氏菌的快速检验。结果表明,该法灵敏度约为2.8 cfu·mL-1,经对205份肉类、蛋、奶及其制品和动物腹泻物、人工污染样品等进行检测,共检出13份阳性样本,与国标(GB 4789.5-2012)方法的检测结果一致。表明建立的荧光PCR方法操作简便、特异性强、灵敏度高,具有良好实用性。%According Shigella ipaH highly conserved gene sequences, probes and primers designed by optimizing the reaction conditions, the establishment of foods of animal origin Shigella real-time PCR method, used in foods of animal origin Shiga rapid test for Shigella. The results showed that the sensitivity of a dual real-time PCR was 2.8 cfu·mL-1. 13 from 205 samples of meat, egg, milk and itsproducts, animal diarrhea materials and artificial contamination samples were positive in a dual real- time PCR assay, which was in accordance with the testing result according to GB 4789.5-2012.The results showed that a dual real- time PCR assay developed in this work was simple, specificity, high sensitivity, good practicality for the detection of Shigella.

  6. Inverse fusion PCR cloning.

    Directory of Open Access Journals (Sweden)

    Markus Spiliotis

    Full Text Available Inverse fusion PCR cloning (IFPC is an easy, PCR based three-step cloning method that allows the seamless and directional insertion of PCR products into virtually all plasmids, this with a free choice of the insertion site. The PCR-derived inserts contain a vector-complementary 5'-end that allows a fusion with the vector by an overlap extension PCR, and the resulting amplified insert-vector fusions are then circularized by ligation prior transformation. A minimal amount of starting material is needed and experimental steps are reduced. Untreated circular plasmid, or alternatively bacteria containing the plasmid, can be used as templates for the insertion, and clean-up of the insert fragment is not urgently required. The whole cloning procedure can be performed within a minimal hands-on time and results in the generation of hundreds to ten-thousands of positive colonies, with a minimal background.

  7. Use of Droplet Digital PCR for Estimation of Fish Abundance and Biomass in Environmental DNA Surveys

    OpenAIRE

    Doi, Hideyuki; Uchii, Kimiko; Takahara, Teruhiko; Matsuhashi, Saeko; Yamanaka, Hiroki; Minamoto, Toshifumi

    2015-01-01

    An environmental DNA (eDNA) analysis method has been recently developed to estimate the distribution of aquatic animals by quantifying the number of target DNA copies with quantitative real-time PCR (qPCR). A new quantitative PCR technology, droplet digital PCR (ddPCR), partitions PCR reactions into thousands of droplets and detects the amplification in each droplet, thereby allowing direct quantification of target DNA. We evaluated the quantification accuracy of qPCR and ddPCR to estimate sp...

  8. Effect of Cage-Wash Temperature on the Removal of Infectious Agents from Caging and the Detection of Infectious Agents on the Filters of Animal Bedding-Disposal Cabinets by PCR Analysis

    OpenAIRE

    Compton, Susan R; Macy, James D

    2015-01-01

    Efficient, effective cage decontamination and the detection of infection are important to sustainable biosecurity within animal facilities. This study compared the efficacy of cage washing at 110 and 180 °F on preventing pathogen transmission. Soiled cages from mice infected with mouse parvovirus (MPV) and mouse hepatitis virus (MHV) were washed at 110 or 180 °F or were not washed. Sentinels from washed cages did not seroconvert to either virus, whereas sentinels in unwashed cages seroconvert...

  9. " Animal, trop animal "

    OpenAIRE

    Potestà, Andréa

    2010-01-01

    Dans la tradition philosophique, on trouve plusieurs définitions de l’homme. La célèbre définition aristotélicienne, zoon logon echon (animal doué du langage ou animal rationnel) fournit le paradigme ainsi que la méthode de toutes les définitions successives. Il s’agit d’ajouter au vivant, à l’animal, quelque chose d’autre, quelque chose de plus, qui permette de le caractériser et le fasse entendre comme différent des bêtes. Cette diversité peut être conçue différemment : en tant qu’élévation...

  10. Real-Time PCR

    Science.gov (United States)

    Evrard, A.; Boulle, N.; Lutfalla, G. S.

    Over the past few years there has been a considerable development of DNA amplification by polymerase chain reaction (PCR), and real-time PCR has now superseded conventional PCR techniques in many areas, e.g., the quantification of nucleic acids and genotyping. This new approach is based on the detection and quantification of a fluorescent signal proportional to the amount of amplicons generated by PCR. Real-time detection is achieved by coupling a thermocycler with a fluorimeter. This chapter discusses the general principles of quantitative real-time PCR, the different steps involved in implementing the technique, and some examples of applications in medicine. The polymerase chain reaction (PCR) provides a way of obtaining a large number of copies of a double-stranded DNA fragment of known sequence. This DNA amplification technique, developed in 1985 by K. Mullis (Cetus Corporation), saw a spectacular development over the space of a few years, revolutionising the methods used up to then in molecular biology. Indeed, PCR has many applications, such as the detection of small amounts of DNA, cloning, and quantitative analysis (assaying), each of which will be discussed further below.

  11. Amazing Animals

    Science.gov (United States)

    Al-Kuwari, Najat Saad

    2007-01-01

    "Animals" is a three-part lesson plan for young learners with a zoo animal theme. The first lesson is full of activities to describe animals, with Simon Says, guessing games, and learning stations. The second lesson is about desert animals, but other types of animals could be chosen depending on student interest. This lesson teaches…

  12. Estudio comparativo de un PCR anidado, ELISA y AGID en la detección del virus de la leucosis bovina en muestras de suero, sangre y leche Comparative study of nested PCR, ELISA and AGID tests in the detection of bovine leukaemia virus infection in serum, blood and milk samples

    Directory of Open Access Journals (Sweden)

    R Felmer

    2006-01-01

    Full Text Available Se evaluaron distintos métodos actualmente disponibles para el diagnóstico de la infección por el virus de la leucosis bovina (VLB. Los métodos empleados fueron AGID en suero, ELISA en muestras de suero y leche y PCR en linfocitos sanguíneos. De un total de 126 animales analizados, AGID identificó un menor número de animales positivos (75 comparado con las pruebas PCR y ELISA aplicadas en muestras de suero y leche (100. Tres animales positivos a AGID fueron negativos a PCR y 28 de las 51 muestras negativas a AGID fueron positivas mediante PCR. La sensibilidad diagnóstica de PCR con respecto a AGID fue de 96%, mientras que la especificidad fue de 45% (kappa 0,45. Todos los animales positivos a AGID fueron también positivos a ELISA aplicado tanto en suero como en leche, mientras que 25 animales negativos a AGID fueron consignados como positivos a ELISA, en ambas muestras biológicas. De esta forma, la sensibilidad diagnóstica de ELISA respecto a AGID fue de un 100%, mientras que la especificidad fue de 51% (kappa 0,55. La menor sensibilidad observada de AGID no es debido a reacciones falso positivas de ELISA y PCR, sino más bien a una mayor sensibilidad de estas últimas, lo que sugiere reconsiderar la utilización del método AGID en aquellos países en que aún se utiliza como método oficial en los programas de erradicación de leucosis.Different methods available for the detection of bovine leukaemia virus (BLV infection were evaluated. The methods evaluated were AGID in serum, ELISA in serum and milk, and PCR in blood lymphocytes. The AGID test identified a smaller number of positive animals (75/126 compared to PCR and ELISA tests (100/126. Three positive animals by AGID were negative by PCR and 28 of the 51 negative samples by AGID were positive by PCR. The sensitivity of PCR with respect to AGID was 96%, whereas the specificity was 45% (kappa 0.45. All positive animals by AGID were also positive by ELISA in serum and milk samples

  13. Overlap extension PCR cloning.

    Science.gov (United States)

    Bryksin, Anton; Matsumura, Ichiro

    2013-01-01

    Rising demand for recombinant proteins has motivated the development of efficient and reliable cloning methods. Here we show how a beginner can clone virtually any DNA insert into a plasmid of choice without the use of restriction endonucleases or T4 DNA ligase. Chimeric primers encoding plasmid sequence at the 5' ends and insert sequence at the 3' ends are designed and synthesized. Phusion(®) DNA polymerase is utilized to amplify the desired insert by PCR. The double-stranded product is subsequently employed as a pair of mega-primers in a PCR-like reaction with circular plasmids. The original plasmids are then destroyed in restriction digests with Dpn I. The product of the overlap extension PCR is used to transform competent Escherichia coli cells. Phusion(®) DNA polymerase is used for both the amplification and fusion reactions, so both steps can be monitored and optimized in the same way. PMID:23996437

  14. Antigenic typing of canine parvovirus using differential PCR.

    Science.gov (United States)

    Kaur, Gurpreet; Chandra, Mudit; Dwivedi, P N; Sharma, N S

    2014-12-01

    Canine parvovirus (CPV) is an enteric pathogen causing hemorrhagic enteritis in pups of 3-6 months of age and is mainly transmitted via feco-oral route. In the present study, a total of 85 animals rectal swabs suspected of CPV were tested using a PCR, nested PCR and a newly designed differential PCR. Using PCR 7 (8.23 %) animals were positive whereas 39 (45.88 %) were positive by using nested PCR and 40 (47.05 %) were positive for either one or more than one antigenic types of CPV using differential PCR. Using differential PCR it was found that CPV-2a and CPV-2b were the most prevailing antigenic types. Also it was found that dogs that were vaccinated too yielded positive CPV indicating a possible presence of additional CPV antigenic types. Thus, the primers used in differential PCR can be used in a single PCR reaction to detect various antigenic types of CPV. PMID:25674626

  15. 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.

  16. Identification of Fel ursi and Cattle and Pig Bile Juices by speciesspecific PCR and PCR-RFLP

    Directory of Open Access Journals (Sweden)

    Ki-Rok Kwon

    2009-03-01

    Full Text Available Objective : This study developed species-specific PCR and PCR-RFLP to detect the adulteration of Fel ursi products with cattle and pig bile juices. Methods : All the primers for PCR and PCR-RFLP in this study were designed based on nucleotide sequences of cytochrome b genes in the mitochondria. Results : The species-specific PCR amplified a DNA fragment of 214, 214, 295, and 167 bp from Felursi product, bear fur, cattle bile juice, and pig bile juice, respectively. The survey using the speciesspecific PCR indicated that some of commercial Fel ursi products were adulterated with cattle and pig bile juices. PCR-RFLP using the restriction endonucleases, HaeIII and HinfI enabled differentiation among Fel ursi product, cattle bile juice, and pig bile juice. Bear furs from two animals showed variations in PCR-RFLP patterns with HaeIII. Discussion : The detection methods of the species-specific PCR and PCR-RFLP could be useful in eliminating adulterated Fel ursi products from the market.

  17. 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

    2016-01-01

    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. PMID:27213452

  18. Colony screening by PCR

    OpenAIRE

    sprotocols

    2014-01-01

    Author: Matt Lewis ### Notes This is the fastest way to screen bacterial colonies. Our PCR machine takes 24 tubes so I routinely screen 22 colonies + 1 negative + 1 positive control. ### Choosing the primers Ideally you want a primer pair that can only work if the correct construct is present eg. a vector flanking primer and a gene specific primer. However, this may not allow you a positive control (essential) so you might have to use both vector flanking primers instead. If y...

  19. 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....

  20. Animal Farm

    Institute of Scientific and Technical Information of China (English)

    徐蓉蓉

    2015-01-01

    This essayfirst 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.

  1. Animal Bites

    Science.gov (United States)

    Wild animals usually avoid people. They might attack, however, if they feel threatened, are sick, or are protecting their ... or territory. Attacks by pets are more common. Animal bites rarely are life-threatening, but if they ...

  2. Animal Bites

    Science.gov (United States)

    ... and complications from bites Never pet, handle, or feed unknown animals Leave snakes alone Watch your children closely around animals Vaccinate your cats, ferrets, and dogs against rabies Spay or neuter ...

  3. 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.

  4. Animal ethics

    OpenAIRE

    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 possibl...

  5. Quadruped Animation

    OpenAIRE

    Skrba, Ljiljana; Reveret, Lionel; Hétroy, Franck; Cani, Marie-Paule; O'Sullivan, Carol

    2008-01-01

    Films like Shrek, Madagascar, The Chronicles of Narnia and Charlotte's web all have something in common: realistic quadruped animations. While the animation of animals has been popular for a long time, the technical challenges associated with creating highly realistic, computer generated creatures have been receiving increasing attention recently. The entertainment, education and medical industries have increased the demand for simulation of realistic animals in the computer graphics area. In...

  6. Thin Animals

    OpenAIRE

    Johnston, D.

    1998-01-01

    Lattice animals provide a discretized model for the theta transition displayed by branched polymers in solvent. Exact graph enumeration studies have given some indications that the phase diagram of such lattice animals may contain two collapsed phases as well as an extended phase. This has not been confirmed by studies using other means. We use the exact correspondence between the q --> 1 limit of an extended Potts model and lattice animals to investigate the phase diagram of lattice animals ...

  7. Trypanosoma spp. in Swedish game animals

    OpenAIRE

    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...

  8. 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

  9. 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...

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

    OpenAIRE

    Danielle C. Leal; Cláudio R. Madruga; Paulo F. de Matos; Bárbara M. P. da S. Souza; Carlos R. Franke

    2011-01-01

    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 equ...

  11. Polimeraz Zincir Reaksiyonu (PCR) Optimizasyonu

    OpenAIRE

    KAHYA, Serpil; BUYUKCANGAZ, Esra; Carli, K. Tayfun

    2013-01-01

    Polymerase chain reaction (PCR), a deoxyribonucleic acid (DNA) that lies between two known chain enzymatically amplify a specific DNA region as an in vitro technique becoming common everyday. PCR method, used for many purposes such as diagnosis, epidemiology and studies to determine the amount of DNA, are still under development. Microbiology is taking a significant proportion in PCR usage, like innovations of application in other fields. Furthermore, PCR is the fundamental molecular method t...

  12. 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.

  13. Animal research

    DEFF Research Database (Denmark)

    Olsson, I.A.S.; Sandøe, Peter

    2012-01-01

    in science (as in any other human use that is not also in the animals’ best interest). These views are not compatible, and since all three views in more or less pure form are found in modern Western societies, use of animals for research is bound to cause controversy. However, there may be room for some kind......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...... 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...

  14. 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...

  15. [Transgenic animals and animal welfare

    Science.gov (United States)

    Reinhardt, Christoph

    1998-01-01

    Under the pressure of a public vote in Switzerland (7 June 1998) on an initiative to ban the production, use and patenting of transgenic animals, their value for biomedical research and development is intensely debated. In addition, the Swiss legislation has adopted (1992) a constitutional obligation to "take into account the dignity of creatures". The term "dignity of creatures", however, can be interpreted in anthropocentric or biocentric ways. The government has now formulated the legal implications of this term for transgenic animals and plants in various laws including the animal and environmental protection laws. This paper gives arguments for a fair evaluation of trangenic animals from an animal welfare point of view where not only the costs of animal suffering must be considered but also the probability of potential benefit for man. A self-confident research community should allow such an evaluation procedure even in view of an outcome which could ban many uses of transgenic animals PMID:11208266

  16. Método para la detección, identificación y cuantificación de Peronospora arborescens por PCR cuantitativa en tiempo real

    OpenAIRE

    Jiménez-Díaz, Rafael M.; Muñoz Ledesma, Francisco Javier; Montes Borrego, Miguel; Landa, Blanca B.

    2009-01-01

    Método para la detección, identificación y cuantificación de Peronospora arborescens por PCR cuantitativa en tiempo real. El método para la cuantificación de Peronospora arborescens por PCR cuantitativa (qPCR) en una muestra biológica, comprende extraer el ADN contenido en dicha muestra biológica y amplificarlo mediante qPCR. De aplicación en la cuantificación de P. arborescens.

  17. 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-

  18. 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....

  19. Animal cytomegaloviruses.

    OpenAIRE

    Staczek, J.

    1990-01-01

    Cytomegaloviruses are agents that infect a variety of animals. Human cytomegalovirus is associated with infections that may be inapparent or may result in severe body malformation. More recently, human cytomegalovirus infections have been recognized as causing severe complications in immunosuppressed individuals. In other animals, cytomegaloviruses are often associated with infections having relatively mild sequelae. Many of these sequelae parallel symptoms associated with human cytomegalovir...

  20. ANIMAL code

    International Nuclear Information System (INIS)

    This report describes ANIMAL, a two-dimensional Eulerian magnetohydrodynamic computer code. ANIMAL's physical model also appears. Formulated are temporal and spatial finite-difference equations in a manner that facilitates implementation of the algorithm. Outlined are the functions of the algorithm's FORTRAN subroutines and variables

  1. 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…

  2. 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

  3. Advances in simulation of PCR

    International Nuclear Information System (INIS)

    Polymerase chain reaction (PCR) is an important diagnosis tool in molecular biology, which have been greatly improved by PCR. However, optimizing the experimental conditions is still a problem for PRC. Computer biology can be a solution to this problem. In this paper, developments of the mathematical models for PCA are reviewed. It is believed that this kind of research efforts shall be helpful for optimizing the experimental conditions and providing guidance for the biologists and understanding the mechanism of PCR. (authors)

  4. Animal learning.

    Science.gov (United States)

    Castro, Leyre; Wasserman, Edward A

    2010-01-01

    Pavlov and Thorndike pioneered the experimental study of animal learning and provided psychologists with powerful tools to unveil its underlying mechanisms. Today's research developments and theoretical analyses owe much to the pioneering work of these early investigators. Nevertheless, in the evolution of our knowledge about animal learning, some initial conceptions have been challenged and revised. We first review the original experimental procedures and findings of Pavlov and Thorndike. Next, we discuss critical research and consequent controversies which have greatly shaped animal learning theory. For example, although contiguity seemed to be the only condition that is necessary for learning, we now know that it is not sufficient; the conditioned stimulus (CS) also has to provide information about the occurrence of the unconditioned stimulus (US). Also, animals appear to learn different things about the same stimuli when circumstances vary. For instance, when faced with situations in which the meaning of a CS changes, as in the case of acquisition and later extinction, animals seem to preserve the original knowledge (CS-US) in addition to learning about the new conditions (CS-noUS). Finally, we discuss how parallels among Pavlovian conditioning, operant conditioning, and human causal judgment suggest that causal knowledge may lie at the root of both human and animal learning. All of these empirical findings and theoretical developments prove that animal learning is more complex and intricate than was once imagined. Copyright © 2009 John Wiley & Sons, Ltd. For further resources related to this article, please visit the WIREs website. PMID:26272842

  5. Digital droplet PCR on disk.

    Science.gov (United States)

    Schuler, Friedrich; Trotter, Martin; Geltman, Marcel; Schwemmer, Frank; Wadle, Simon; Domínguez-Garrido, Elena; López, María; Cervera-Acedo, Cristina; Santibáñez, Paula; von Stetten, Felix; Zengerle, Roland; Paust, Nils

    2016-01-01

    Existing systems for digital droplet PCR (ddPCR) either suffer from low integration or are difficult to introduce to mass fabrication. Here we present an integrated system that is compatible to mass fabrication and combines emulsification, PCR, and fluorescence readout in a single chamber within a disposable cartridge (disk). Droplets are generated by injecting the sample into fluorinated oil via centrifugal step emulsification. The resulting emulsion is aligned in the PCR and readout zone by capillary action. During thermocycling, gas bubbles generated by degassing are removed by capillary driven transport through tapered regions in the PCR chamber. Thereby, the positioning of the emulsion within the readout zone of the PCR chamber is ensured at any time and no bubbles are present during readout. Manual handling of the disk solely requires pipetting of oil and PCR mix into the inlet structures, placing the disk into the thermocycler and subsequently into a microarray scanner. The functionality of the ddPCR process chain is demonstrated by quantitative detection of the cystic fibrosis causing mutation p.Phe508del, which is of interest for non-invasive prenatal testing (NIPT). The mutation was detected in a concentration range spanning four orders of magnitude. We envision that this work will lay the base for the development of highly integrated sample-to-digital-answer PCR systems that can be employed in routine clinical diagnosis. PMID:26610263

  6. 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.

  7. Adsorción de boro mediante perlas de alginato

    OpenAIRE

    Seira Ibáñez, Juana

    2008-01-01

    En este proyecto se propone la técnica de la adsorción mediante la utilización de polímeros naturales para eliminar el boro de residuos industriales, puesto que estos residuos presentan una gran problemática medioambiental. El polímero elegido para realizar la adsorción en este estudio es el alginato. Para poder trabajar en estado sólido se transforma el alginato de sodio, que es soluble en agua, en gel mediante la fabricación de las perlas de alginato de calcio. (Se utiliza...

  8. Desarrollo comunicación Alfa Arduino mediante Scada

    OpenAIRE

    Trallero Calvo, Jorge

    2015-01-01

    El siguiente proyecto pretende crear un sistema IoT (Internet of Things) girando en torno a Arduino, demostrando de este modo la facilidad de creación de proyectos y fiabilidad que nos aporta Arduino. Mediante una comunicación TCP/IP se procede a crear un sistema el cual es capaz de registrar valores a una cierta distancia mediante protocolos de procesos y comunicación (Telemedida) y de este modo monitorizar la temperatura en un punto determinado, sin estar limitado por la t...

  9. Animal performance

    OpenAIRE

    Abaye, A. O. (Azenegashe Ozzie); Rotz, Jonathan Daniel; Scaglia Alonso, Guillermo, 1963-; Fike, John Herschel; Smith, Ray Lee, 1962-

    2009-01-01

    Any forage crop that stretches the grazing season by providing additional feed in early spring, mid-summer, and late fall will provide the livestock producer with lower feed costs and boost animal performance.

  10. Animation & Neurocinematics*

    DEFF Research Database (Denmark)

    Carpe Pérez, Inmaculada Concepción

    2016-01-01

    , indeed, can be considered a social/ emotional learning media, which goes beyond the limitations of live action movies. This is due to the diversity of techniques, and its visual plasticity that constructs the impossible. Animators are not real actors but more like the midwife who brings the anima...... 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...... into aliveness, which requires knowing how emotions work. Ed Hooks as an expert in training animators and actors, always remarks: “emotions tend to lead to action”. In this paper we want to argue that by producing animated films, as we watch them, cause a stronger effect, not only in our brains, but also in our...

  11. Groundwater animals

    OpenAIRE

    Maurice, Louise; Bloomfield, John; Robertson, Anne; Allen, Debbie

    2010-01-01

    Groundwater animals are adapted to live in environments with no light and limited nutrients, They can provide insights into fundamental questions of evolution, ecology and biodiversity. They also have an important role to play in informing the reconstruction of past changes in geomorphology and climate, and can be used for characterising aquifers. The BGS is undertaking a systematic survey of selected areas and lithologies in the UK where groundwater animals have not been inves...

  12. Turismo, Anime y Comunidad, ¿por qué ahora? : Potencial del turismo mediante los contenidos de anime

    OpenAIRE

    Yamamura, Takayoshi

    2010-01-01

    2 de marzo del 2010, Curso de Capacitacion de JICA, Region Latinoamericana, Curso “Desarrollo de Turismo Regional Sostenible” Material Didactico (2010年3月2日 JICA札幌研修事業「中南米地域 持続可能な地域観光開発コース 教材)

  13. From the 'PCR' function to the 'PCR' profession

    International Nuclear Information System (INIS)

    After having recalled the legal context concerning the appointment and training of a radiation protection expert (PCR for 'personne competente en radioprotection'), the author outlines that the PCR's role has notably evolved: his function is now of primary importance in the company and his activity does not correspond to the legal framework any longer. Moreover, with the application of a European directive, some small establishments possessing ionizing radiation sources are disadvantaged, and the PCR is now facing an increasing number of missions and tasks. The author gives a list of them and assesses a needed time of 146 days per year: this means PCRs cannot have an other activity within their company

  14. Biotecnologia animal

    OpenAIRE

    Luiz Lehmann Coutinho; Millor Fernandes do Rosário; Erika Cristina Jorge

    2010-01-01

    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 candi...

  15. 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...

  16. 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?

  17. Transgenic Animals.

    Science.gov (United States)

    Jaenisch, Rudolf

    1988-01-01

    Describes three methods and their advantages and disadvantages for introducing genes into animals. Discusses the predictability and tissue-specificity of the injected genes. Outlines the applications of transgenic technology for studying gene expression, the early stages of mammalian development, mutations, and the molecular nature of chromosomes.…

  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. Multiplex PCR:a powerful and affordable tool for laboratory and field analysis in developing countries

    Institute of Scientific and Technical Information of China (English)

    Mohamed; O; Ahmed

    2014-01-01

    To the editor,Since the introduction of multiplex-PCR(mPCR)in1988,this technique has emerged as a highly efficient and sensitive molecular tool for nucleic acid-based diagnosis and monitoring,it is applicable to a broad range of physiological,metabolic and infectious conditions affecting human,animal

  20. 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......; 10 countries). Conclusions: This results show that although PCR ribotype 078 is often reported as the major animal C. difficile type, especially in pigs, the variability of strains in pigs and other animal hosts is substantial. Most common human PCR ribotypes (014/020 and 002) are also among most...... prevalent 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....

  1. 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

  2. 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.

  3. Animal facilities

    International Nuclear Information System (INIS)

    The animal facilities in the Division are described. They consist of kennels, animal rooms, service areas, and technical areas (examining rooms, operating rooms, pathology labs, x-ray rooms, and 60Co exposure facilities). The computer support facility is also described. The advent of the Conversational Monitor System at Argonne has launched a new effort to set up conversational computing and graphics software for users. The existing LS-11 data acquisition systems have been further enhanced and expanded. The divisional radiation facilities include a number of gamma, neutron, and x-ray radiation sources with accompanying areas for related equipment. There are five 60Co irradiation facilities; a research reactor, Janus, is a source for fission-spectrum neutrons; two other neutron sources in the Chicago area are also available to the staff for cell biology studies. The electron microscope facilities are also described

  4. [Dangerous animals].

    Science.gov (United States)

    Hasle, Gunnar

    2002-06-30

    As travellers seek ever more exotic destinations they are more likely to encounter dangerous animals. Compared to risks such as AIDS, traffic accidents and malaria, the risk is not so great; many travellers are, however, concerned about this and those who give pre-travel vaccines and advice should know something about it. This article is mainly based on medical and zoological textbooks. Venomous stings and bites may be prevented by adequate clothing and by keeping safe distance to the animals. Listening to those who live in the area is of course important. Travellers should not carry antisera with them, but antisera should be available at local hospitals. It should be borne in mind that plant eaters cause just as many deaths as large predators. In some cases it is necessary to carry a sufficiently powerful firearm. PMID:12555616

  5. 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.

  6. A study of PCR inhibition mechanisms using real time PCR.

    Science.gov (United States)

    Opel, Kerry L; Chung, Denise; McCord, Bruce R

    2010-01-01

    In this project, real time polymerase chain reaction (PCR) was utilized to study the mechanism of PCR inhibition through examination of the effect of amplicon length, melting temperature, and sequence. Specifically designed primers with three different amplicon lengths and three different melting temperatures were used to target a single homozygous allele in the HUMTH01 locus. The effect on amplification efficiency for each primer pair was determined by adding different concentrations of various PCR inhibitors to the reaction mixture. The results show that a variety of inhibition mechanisms can occur during the PCR process depending on the type of co-extracted inhibitor. These include Taq inhibition, DNA template binding, and effects on reaction efficiency. In addition, some inhibitors appear to affect the reaction in more than one manner. Overall we find that amplicon size and melting temperature are important in some inhibition mechanisms and not in others and the key issue in understanding PCR inhibition is determining the identity of the interfering substance. PMID:20015162

  7. Animal Drug Safety FAQs

    Science.gov (United States)

    ... Vaccines, Blood & Biologics Animal & Veterinary Cosmetics Tobacco Products Animal & Veterinary Home Animal & Veterinary Safety & Health Frequently Asked Questions Animal Drug Safety Frequently Asked Questions Share Tweet Linkedin ...

  8. Control de velocidad mediante relación voltajefrecuencia

    OpenAIRE

    Alfonso Álzate; Duberney Murillo Yarce; Marcela González Valencia

    2011-01-01

    Se presenta la aplicación de un control digital utilizando un DSP, para controlar la velocidad de un motor de inducción trifásico en lazo cerrado mediante la técnica de control voltaje-frecuencia. También se presenta un panorama general de las técnicas1 de control escalar y vectorial, analizando en detalle la técnica escalar voltaje-frecuencia.

  9. Detección molecular de las translocaciones más comunes en leucemia aguda mediante rt-pcr

    OpenAIRE

    Guevara G.; García L.

    2011-01-01

    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.

  10. 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.

  11. 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

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

    OpenAIRE

    Giovambattista Guillermo; Muñoz Flórez Jaime Eduardo; Posso Terranova Andres Mauricio; Hernández Herrera Darwin Yovanny; Antonio Benavides Javier

    2011-01-01

    Se evaluó la presencia del virus de la leucosis bovina (VLB) en 360 muestras de ADN de oc%o razas bovinas criollas: Blanco (re)inegro  (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 (H)J Para la detección del proKvirus se amplificó una región del gen ...

  13. 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...

  14. 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.

  15. Animated war

    DEFF Research Database (Denmark)

    Frølunde, Lisbeth

    2012-01-01

    in production: Gzim Rewind (Sweden, 2011) by Knutte Wester, and In-World War (USA, expected 2011) by DJ Bad Vegan. These films have themes of war and include film scenes that are ‘machinima’ (real-time animation made in 3D graphic environments) within live action film scenes. Machinima harnesses...... DIY multimedia storytellers explore new ways to tell and to ‘animate’ stories. The article contains four parts: introduction to machinima and the notions of resemiosis and authorial practice, presentation of DIY filmmaking as a practice that intertwines with new networked economics, analysis...

  16. Molecular detection of Trypanosoma infection: PCR and PCR-ELISA techniques

    International Nuclear Information System (INIS)

    Fast, clear and easy diagnostic methods allow accurate detection of Trypanosomes in human and animal species enabling cost effective treatment and prevention of outbreaks within an epidemic. Diagnostic methods should preferably be usable in the field. Presently many diagnostic field methods are time consuming and their limit for detection of parasites should be decreased. Molecular methods offer the promise of detecting lower numbers of parasites, faster. One focus of this RCM was to develop fast and easy diagnostic methods detecting lower infection levels. The original ideas involved PCR-ELISA and based on our previous experience, we confirm that PCR-ELISA is indeed a powerful method able to detect minor quantities of target DNA, i.e. very low parasite copy numbers, down to a single parasite. We also point out that the method has several pitfalls and should therefore be applied with great care. During the first research coordination meeting in Belgium, we presented results with RT-PCR-ELISA and the controls we used to make our results reliable. This manuscript summarizes these data. (author)

  17. 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

    ; 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...... results. Finally, the Microsporum PCR was positive for 10/10 guinea pig specimens from infected animals but for 0/2 of the control animal samples. The evaluation of the two PCR tests indicated excellent sensitivity and specificity.......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...

  18. Animal Intuitions.

    Science.gov (United States)

    Kaebnick, Gregory E

    2016-07-01

    As described by Lori Gruen in the Perspective column at the back of this issue, federally supported biomedical research conducted on chimpanzees has now come to an end in the United States, although the wind-down has taken longer than expected. The process began with a 2011 Institute of Medicine report that set up several stringent criteria that sharply limited biomedical research. The National Institutes of Health accepted the recommendations and formed a committee to determine how best to implement them. The immediate question raised by this transition was whether the IOM restrictions should be extended in some form to other nonhuman primates-and beyond them to other kinds of animals. In the lead article in this issue, Anne Barnhill, Steven Joffe, and Franklin Miller consider the status of other nonhuman primates. PMID:27417859

  19. 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.

  20. Online exercise for the design and simulation of PCR and PCR-RFLP experiments

    OpenAIRE

    San Millán Gutiérrez, Rosario María; Martínez Ballesteros, Ilargi; Rementeria Ruiz, Aitor Domingo; Garaizar Candina, Javier; Bikandi Bikandi, Joseba

    2013-01-01

    [EN] Background: Polymerase Chain Reaction (PCR) and Restriction Fragment Length Polymorphism of PCR products (PCR-RFLP) are extensively used molecular biology techniques. An exercise for the design and simulation of PCR and PCR-RFLP experiments will be a useful educational tool. Findings: An online PCR and PCR-RFLP exercise has been create that requires users to find the target genes,compare them, design primers, search for restriction endonucleases, and finally to simulate the experiment...

  1. 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)

  2. 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.

  3. Nested PCR for Specific Diagnosis of Taenia solium Taeniasis▿

    OpenAIRE

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

    2007-01-01

    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 devel...

  4. Comparison of droplet digital PCR and qPCR for the quantification of Shiga toxin-producing Escherichia coli in bovine feces

    OpenAIRE

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

    2016-01-01

    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 ...

  5. Comparison of toxicity neutralization-, ELISA- and PCR tests for typing of Clostridium perfringens and detection of the enterotoxin gene by PCR

    DEFF Research Database (Denmark)

    Møller, Kristian; Ahrens, Peter

    1996-01-01

    constructed and served as a control for inhibition of the PCR(beta) test. The enterotoxin gene was not in any of 95 Danish Clostridium perfringens field isolates. This indicates that the C. perfringens enterotoxin is not involved in diarrhoea in certain animal species from this area. The origin of enterotoxin-positive......A polymerase chain reaction (PCR) was developed for the specific amplification of a part of each of the five Clostridium perfringens toxin genes: alpha (alpha), beta (beta), epsilon (epsilon), iota (iota), and enterotoxin (CPE). While the toxicity neutralization test (TNT) only showed limited...... ability to detect the or toxin, the lecithinase test and PCR test (PCR(alpha)) concordantly detected the ct toxin and the alpha toxin gene, respectively. A monoclonal enzyme linked immunosorbent assay (ELISA) and a PCR(beta) test were compared and were in accordance for the detection of the beta toxin...

  6. PCR Based Diagnosis of Fungal Diseases in Dogs

    Directory of Open Access Journals (Sweden)

    Idress Hamad Attitalla

    2012-01-01

    Full Text Available Interaction with animals provide necessary companionship and helps people to live better life by reducing risk of many health problems, improved fitness and act as a source of social enjoyment. In modern society there is a substantial increase in the number of dogs adopted as pet which also raises the concerns regarding the transmission of infections from dog to their owners and vice versa. Early diagnosis of infected dogs could prevent the owners from these infections. In last decade, PCR has proven its potential applications as an important diagnostic tool but these applications are mainly limited to the diagnosis of human diseases and very little work has been done on animals. Development of PCR assays for detection of commonly found canine fungal infections (aspergillosis, blastomycosis, coccidioidomycosis and cryptococcosis is of utmost importance in order to ensure the early and accurate diagnosis of infection. Although, a lot of research is being done on molecular diagnosis of animal infectious disease but most of these newly developed assays are either not well optimized or only suitable for laboratory studies. On the basis of reviewed literature it can be concluded that more research work is required to develop an efficient (rapid, sensitive and cost effective PCR assays for the diagnosis of canine fungal infections.

  7. Tratamiento del síndrome subacromial mediante acromioplastia abierta

    OpenAIRE

    Arenas Planelles, Antonio; Garbayo Marturet, Antonio Jesús; Ayala Palacios, Higinio; Arenas Miquélez, A.

    2003-01-01

    Se presenta una serie de 161 casos de síndrome subacromial tratados quirúrgicamente en nuestro Servicio mediante acromioplastia abierta y reconstrucción del manguito en los casos en que existía ruptura del mismo. El estudio se ha realizado de forma retrospectiva, evaluándose una serie de variables clínicas, radiológicas y quirúrgicas, y algunos parámetros para valorar los resultados. Tras la intervención, con un tiempo de seguimiento medio de 45 meses, el 82% de los pacientes se encontraban s...

  8. RECURSOS NO CONVENCIONALES SUSCEPTIBLES DE SER EXPLOTADOS MEDIANTE FRACKING

    OpenAIRE

    Jódar Abellán, Antonio

    2014-01-01

    En el presente trabajo se analizan en detalle los recursos de origen no convencional existentes en el contexto internacional, europeo, nacional y regional, que pueden ser explotados mediante fracking. Así mismo, se mencionará someramente la legislación aplicable a la fracturación hidráulica y los impactos, tanto beneficiosos como perjudiciales, que ésta puede acarrear. El estudio finaliza con una propuesta de posibles lugares en la Península Ibérica donde dicha técnica reporte mayores benefic...

  9. Verificando diseños BON mediante Alloy

    OpenAIRE

    Castro, Pablo Francisco; Ponzio, Pablo Daniel; Demasi, Ramiro Adrián; Baum, Gabriel Alfredo

    2005-01-01

    En este artículo presentamos una técnica para traducir diseños estructurales expresados en el lenguaje BON, al lenguaje formal Alloy. En donde, la principal ventaja de la traducción es que puede realizarse automáticamente mediante herramientas de software. Adicionalmente, esta metodología puede ser usada para validar propiedades sobre los diseños utilizando el Alloy Analyzer. Para finalizar, mostramos la aplicación a un caso de estudio de Darwin Tool, una herramienta que implem...

  10. Control domótico remoto de vivienda mediante smartphone

    OpenAIRE

    ALEIXANDRE TUDO, BERNARDO

    2013-01-01

    La tecnología forma parte de la sociedad actual y de la vida cotidiana de las personas. Los últimos avances tecnológicos han simplificado nuestra vida y han provocado muchos cambios en las diversas áreas de la sociedad. Una de estas áreas es sin duda la automatización de la vivienda. Los nuevos aparatos electrónicos nos permiten tener acceso a las comunicaciones en cualquier lugar y a cualquier hora. La entrada de internet móvil en el mercado mediante los Smartphone y su ráp...

  11. Animated nature

    International Nuclear Information System (INIS)

    Animated nature is educational-training project pronounced by the Slovak Environmental Agency (SAZP) in cooperation with Field Studies Council form Great Britain and financial support of Darwin Initiative and Slovensky plynarensky priemysel, s.p. In the present time this is ultimate and the most successful children's project aimed on mapping and protection of biodiversity in Europe. Activity in project is spare-time and therefore is voluntary. The interest territory is a natural as well as cultural landscape in vicinity of a school or other organisation, habitation and so on. In the project work schoolchildren at the age from 10 till 15 years. Leaders of work-groups are student of secondary schools and universities, teachers, professional workers of state and non-governmental organisation and parents. In one group works approximately 10 children. Each group which has send to SAZP result of biodiversity mapping, cost free obtained data base CD - Detske mapy biodiverzity (Children's maps of biodiversity) and so they were informed about results of all groups frame: within the frame of Slovakia. Results of activities of this project in 2001-2004 and perspectives for 2005-2006 years are discussed

  12. 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. PMID:26341923

  13. 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.

  14. Viral Multiplex Quantitative PCR Assays for Tracking Sources of Fecal Contamination▿

    OpenAIRE

    Wolf, Sandro; Hewitt, Joanne; Greening, Gail E.

    2010-01-01

    Human and animal fecal pollution of the environment presents a risk to human health because of the presence of pathogenic viruses and bacteria. To distinguish between human and animal sources of pollution, we designed specific real-time reverse transcription (RT)-PCR assays for human and animal enteric viruses, including norovirus genogroups I, II, and III; porcine adenovirus types 3 and 5; ovine adenovirus; atadenovirus; and human adenovirus species C and F, which are excreted by infected hu...

  15. 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

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

    OpenAIRE

    Alejandro Córdova Izquierdo; Claudio Gustavo Ruiz Lang; Jorge A. Saltijeral Oaxaca; Víctor Xolalpa Campos; Saúl Cortés Suárez; Maximino Méndez Mendoza; Rubén Huerta Crispin; Mary S Córdova Jiménez; Córdova Jiménez, Cristian A.; Eulogio Guerra Liera

    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...

  17. Comparison of Droplet Digital PCR to Real-Time PCR for Quantitative Detection of Cytomegalovirus

    OpenAIRE

    Hayden, R. T.; Gu, Z; Ingersoll, J; Abdul-Ali, D.; Shi, L; Pounds, S.; Caliendo, A. M.

    2013-01-01

    Quantitative real-time PCR (QRT-PCR) has been widely implemented for clinical viral load testing, but a lack of standardization and relatively poor precision have hindered its usefulness. Digital PCR offers highly precise, direct quantification without requiring a calibration curve. Performance characteristics of real-time PCR were compared to those of droplet digital PCR (ddPCR) for cytomegalovirus (CMV) load testing. Tenfold serial dilutions of the World Health Organization (WHO) and the Na...

  18. Animation of Antimicrobial Resistance

    Medline Plus

    Full Text Available ... Radiation-Emitting Products Vaccines, Blood & Biologics Animal & Veterinary Cosmetics Tobacco Products Animal & Veterinary Home Animal & Veterinary Safety & ... Radiation-Emitting Products Vaccines, Blood & Biologics Animal & Veterinary Cosmetics Tobacco Products

  19. Animation of Antimicrobial Resistance

    Medline Plus

    Full Text Available ... Emitting Products Vaccines, Blood & Biologics Animal & Veterinary Cosmetics Tobacco Products Animal & Veterinary Home Animal & Veterinary Safety & Health ... Emitting Products Vaccines, Blood & Biologics Animal & Veterinary Cosmetics Tobacco Products

  20. 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: ...

  1. Claves para acreditar una técnica de PCR : caso práctico: Salmonella spp. en leche y productos lácteos

    OpenAIRE

    Galbany, M.; Lázaro, M.

    2013-01-01

    La validación del protocolo para la detección de Salmonella spp por la técnica de la PCR y su posterior comprobación mediante los controles de aseguramiento de la calidad, nos ha permitido obtener la acreditación en una técnica que ofrece la posibilidad a nuestros clientes de obtener resultados en poco tiempo.

  2. 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

  3. Evaluation of two PCR-based techniques for molecular epidemiology in Finland, a high-endemic area with four sympatric Trichinella species

    OpenAIRE

    Kapel C.M.O.; Oivanen L; La Rosa G.; Mikkonen T.; Pozio E.

    2001-01-01

    Trichinella larvae collected from wildlife, domestic and synanthropic animals in Finland were identified to species by two molecular techniques: Random amplified polymorphic DNA (RAPD) polymerase chain reaction (PCR) and the recently described multiplex PCR. The RAPD-PCR was very sensitive to the sub-optimal preservation muscle larvae and resulting in weak and smeared bands on the gels for such material. However, the same samples yielded easily recognizable bands in the multiplex PCR; this la...

  4. 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

    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-negative on...

  5. 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

  6. Real Time PCR: Principles and Application

    OpenAIRE

    Safie Amini; Seyed-Moayed Alavian; Malek Hossein Ahmadipour

    2005-01-01

    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 ...

  7. 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

  8. PCR and real-time PCR primers developed for detection and identification of Bifidobacterium thermophilum in faeces

    Directory of Open Access Journals (Sweden)

    Mini Raffaella

    2008-10-01

    Full Text Available Abstract Background Culture-independent methods based on the 16S ribosomal RNA molecule are nowadays widely used for assessment of the composition of the intestinal microbiota, in relation to host health or probiotic efficacy. Because Bifidobacterium thermophilum was only recently isolated from human faeces until now, no specific real-time PCR (qPCR assay has been developed for detection of this species as component of the bifidobacterial community of the human intestinal flora. Results Design of specific primers and probe was achieved based on comparison of 108 published bifidobacterial 16S rDNA sequences with the recently published sequence of the human faecal isolate B. thermophilum RBL67. Specificity of the primer was tested in silico by similarity search against the sequence database and confirmed experimentally by PCR amplification on 17 Bifidobacterium strains, representing 12 different species, and two Lactobacillus strains. The qPCR assay developed was linear for B. thermophilum RBL67 DNA quantities ranging from 0.02 ng/μl to 200 ng/μl and showed a detection limit of 105 cells per gram faeces. The application of this new qPCR assay allowed to detect the presence of B. thermophilum in one sample from a 6-month old breast-fed baby among 17 human faecal samples tested. Additionally, the specific qPCR primers in combination with selective plating experiments led to the isolation of F9K9, a faecal isolate from a 4-month old breast-fed baby. The 16S rDNA sequence of this isolate is 99.93% similar to that of B. thermophilum RBL67 and confirmed the applicability of the new qPCR assay in faecal samples. Conclusion A new B. thermophilum-specific qPCR assay was developed based on species-specific target nucleotides in the 16S rDNA. It can be used to further characterize the composition of the bifidobacterial community in the human gastrointestinal tract. Until recently, B. thermophilum was considered as a species of animal origin, but here we

  9. 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

  10. Authentication of Meat Species in Sucuk by Multiplex PCR

    Directory of Open Access Journals (Sweden)

    Osman İrfan İLHAK

    2015-01-01

    Full Text Available The identification of meat species used in meat products is important by reason of economic considerations, religious factors, verification of label, and prevention of unfair-market competition. In this paper, multiplex PCR method was experienced for routine detection of equine (horse and donkey, poultry (chicken and turkey, pig and cattle meat in sucuk (sausage. The primers used for these animals generated specific fragments, and they did not show cross reactions with the DNA from the other genus of animal. After multiplex PCR was successfully optimized, a field study was carried out to investigate the presence of horse, donkey, chicken, turkey and pig meat in 50 sucuks (30 beef and 20 beef + poultry collected from markets. The result of the field study indicated that 23.3% of 30 beef sucuk samples were containing poultry meat. None of the 50 sucuk samples was containing pig meat, but one (2% of the samples generated equine fragment. The present study showed that the multiplex PCR method can be used for routine analysis of meat species identification, verification and control of label information of meat products.

  11. Modelo animal de enfermedades neurodegenerativas, procedimiento de obtención y aplicaciones

    OpenAIRE

    Torres Aleman, Ignacio; Carro, Eva; Trejo, Jose L.; Spuch Calvar, Carlos

    2007-01-01

    Se describe un animal no humano útil como modelo experimental de enfermedades neurodegenerativas. Este modelo animal presenta una alteración de la actividad biológica del receptor del factor trófico IGF-I localizada en las células del epitelio del plexo coroideo de los ventrículos cerebrales. Dicho modelo animal puede obtenerse mediante un proceso de transgénesis. Este modelo animal es útil para el estudio de los mecanismos etiopatogénicos de enfermedades neurodegenera...

  12. Animal welfare assessment

    OpenAIRE

    Vučinić Marijana; Lazić Ivana

    2008-01-01

    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 interpr...

  13. New PCR diagnostic systems for the detection and quantification of porcine cytomegalovirus (PCMV).

    Science.gov (United States)

    Morozov, Vladimir A; Morozov, Alexey V; Denner, Joachim

    2016-05-01

    Pigs are frequently infected with porcine cytomegalovirus (PCMV). Infected adult animals may not present with symptoms of disease, and the virus remains latent. However, the virus may be transmitted to human recipients receiving pig transplants. Recently, it was shown that pig-to-non-human-primate xenotransplantations showed 2 to 3 times lower transplant survival when the donor pig was infected with PCMV. Therefore, highly sensitive methods are required to select virus-free pigs and to examine xenotransplants. Seven previously established PCR detection systems targeting the DNA polymerase gene of PCMV were examined by comparison of thermodynamic parameters of oligonucleotides, and new diagnostic nested PCR and real-time PCR systems with improved parameters and high sensitivity were established. The detection limit of conventional PCR was estimated to be 15 copies, and that of the nested PCR was 5 copies. The sensitivity of the real-time PCR with a TaqMan probe was two copies. An equal efficiency of the newly established detection systems was shown by parallel testing of DNA from sera and blood of six pigs, identifying the same animals as PCMV infected. These new diagnostic PCR systems will improve the detection of PCMV and therefore increase the safety of porcine xenotransplants. PMID:26839086

  14. 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…

  15. Animal rights, animal minds, and human mindreading

    OpenAIRE

    Mameli, M.; Bortolotti, L

    2006-01-01

    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 s...

  16. Animal Protection and Animal 'Rights' in Hungary

    OpenAIRE

    Toth, Zoltan J.

    2012-01-01

    In Hungary, the first Act on Animal Protection, which aimed at handling and respecting animals as living creatures capable of feelings and suffering and thus deserving and entitled to protection, was adopted in 1998. Based on this, the Act contains several regulations which ensure that animals are protected against all possible kinds of avoidable physical or mental harm. Furthermore, it prohibits and imposes sanctions for any treatment that causes animals unnecessary suffering. The present st...

  17. [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. PMID:24660572

  18. 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

    2000-01-01

    Milk and faeces samples from cows with clinical symptoms of paratuberculosis were examined for the presence of Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) by culture and PCR. M. paratuberculosis was cultivated in variable numbers from faeces or intestinal mucosa in eight of 11...... animals. In milk from five cows (all faeces culture positive), we cultivated a few colonies of M. paratuberculosis (<100 CFU per ml). Milk samples from two cows were PCR positive (both animals were faeces culture positive, and one cow was milk culture positive). One cow was culture negative on intestinal...

  19. Chip PCR. I. Surface passivation of microfabricated silicon-glass chips for PCR.

    OpenAIRE

    Shoffner, M A; Cheng, J; Hvichia, G E; Kricka, L J; Wilding, P.

    1996-01-01

    The microreaction volumes of PCR chips (a microfabricated silicon chip bonded to a piece of flat glass to form a PCR reaction chamber) create a relatively high surface to volume ratio that increases the significance of the surface chemistry in the polymerase chain reaction (PCR). We investigated several surface passivations in an attempt to identify 'PCR friendly' surfaces and used those surfaces to obtain amplifications comparable with those obtained in conventional PCR amplification systems...

  20. 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.

  1. 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

  2. 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.

  3. Animation of Antimicrobial Resistance

    Medline Plus

    Full Text Available ... 08 Animation of Antimicrobial Resistance (text version) Arabic Translation - Animation of Antimicrobial Resistance (WMV - 19.2MB) Chinese Translation - Animation of Antimicrobial Resistance (WMV - 19.2MB) French ...

  4. Animation of Antimicrobial Resistance

    Medline Plus

    Full Text Available ... Animal & Veterinary Cosmetics Tobacco Products Animal & Veterinary ... The Food and Drug Administration's (FDA's) Center for Veterinary Medicine (CVM) produced a nine-minute animation explaining how ...

  5. Animation of Antimicrobial Resistance

    Science.gov (United States)

    ... 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 ...

  6. 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 ...

  7. Animation of Antimicrobial Resistance

    Medline Plus

    Full Text Available ... Home Food Drugs Medical Devices Radiation-Emitting Products Vaccines, Blood & Biologics Animal & Veterinary Cosmetics Tobacco Products Animal & ... back Food Drugs Medical Devices Radiation-Emitting Products Vaccines, Blood & Biologics Animal & Veterinary Cosmetics Tobacco Products

  8. Molecular identification of trypanosomatids in wild animals.

    Science.gov (United States)

    Tenório, M S; Oliveira e Sousa, L; Alves-Martin, M F; Paixão, M S; Rodrigues, M V; Starke-Buzetti, W A; Araújo Junior, J P; Lucheis, S B

    2014-06-16

    Diverse wild animal species can be reservoirs of zoonotic flagellate parasites, which can cause pathologic Chagas disease. The present study aimed to detect the natural occurrence of flagellate parasites through direct microscopic examination of the parasites in blood samples and through PCR of whole blood and blood culture (haemoculture) samples from 38 captive and 65 free-living wild animals in the Centre for Conservation of Wild Fauna (CCWF), an area endemic for leishmaniasis. For this study, PCR was accomplished using primers for the ribosomal region (ITS-1) of the flagellate parasites. The amplified fragments were cloned and sequenced to identify DNA of the Trypanosomatid parasite species, observed in blood cultures from 3.9% (04/103) of the animals. Through these techniques, Trypanosoma cruzi was identified in haemoculture samples of the following three free-living species: common agouti (Dasyprocta aguti), white-eared opossum (Didelphis albiventris), and nine-banded armadillo (Dasypus novemcinctus). Furthermore, Trypanosoma minasense was identified in whole blood samples from 01 (0.9%) captive animal (black howler monkey-Alouatta caraya). These results demonstrated the first report of T. cruzi isolation in wild species from the CCWF using blood culture, which can be applied in addition to molecular tools for epidemiological studies and to identify trypanosomatids in wild animals. PMID:24636787

  9. Animation Trends in Education

    OpenAIRE

    Lirong Xiao

    2013-01-01

    In the paper, we give a survey of animation content in education. At present, there is an extensive literature addressing the impact of animation in education and psychology fields. However, in animation field, although some software companies have developed their individual production toolboxes or platforms for animation content in education, there is lack of relevant research from the perspective of animation techniques. This paper first gives a survey of current animation content in educat...

  10. T-linker-specific ligation PCR (T-linker PCR): an advanced PCR technique for chromosome walking or for isolation of tagged DNA ends

    OpenAIRE

    Yuanxin, Yan; Chengcai, An; Li, Li; Jiayu, Gu; Guihong, Tan; Zhangliang, Chen

    2003-01-01

    Dozens of PCR-based methods are available for chromosome walking from a known sequence to an unknown region. These methods are of three types: inverse PCR, ligation-mediated PCR and randomly primed PCR. However, none of them has been generally applied for this purpose, because they are either difficult or inefficient. Here we describe a simple and efficient PCR strategy—T-linker-specific ligation PCR (T-linker PCR) for gene or chromosome walking. The strategy amplifies the template molecules ...

  11. Seeing the animal

    DEFF Research Database (Denmark)

    Harfeld, Jes Lynning; Cornou, Cecile; 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. Assessment of the real-time PCR and different digital PCR platforms for DNA quantification

    OpenAIRE

    Pavšič, Jernej; Žel, Jana; Milavec, Mojca

    2015-01-01

    Digital PCR (dPCR) is beginning to supersede real-time PCR (qPCR) for quantification of nucleic acids in many different applications. Several analytical properties of the two most commonly used dPCR platforms, namely the QX100 system (Bio-Rad) and the 12.765 array of the Biomark system (Fluidigm), have already been evaluated and compared with those of qPCR. However, to the best of our knowledge, direct comparison between the three of these platforms using the same DNA material has not been do...

  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. 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.

  15. Muestras y representatividad en vigilancia epidemiologica mediante sitios centinelas

    Directory of Open Access Journals (Sweden)

    Juan Samaja

    1996-09-01

    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.

  16. 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.

  17. COLECTOR SOLAR CONSTRUIDO MEDIANTE TALADRADO POR FLUENCIA TÉRMICA

    Directory of Open Access Journals (Sweden)

    Víctor Heredia R.

    2005-08-01

    Full Text Available Se diseñó y construyó un sistema de calentamiento de agua residencial, mediante dos colectores solares y un estanque. En el sistema se utilizaron tubos de cobre perforados por taladrado por fluencia térmica (TFT, unidos con soldadura de plata. El sistema funciona por termosifón y se emplea como fluido de calentamiento una mezcla de etilenglicol-agua. El fluido de calentamiento pasa a través de un serpentín de cobre a un estanque de 200 litros. Diariamente se calientan 80 litros de agua a una temperatura máxima de 45º C.A heating equipment for residential water was built, using two solar collectors and one isolated reservoir. The system uses to copper tubes perforated by thermal flow drilling (TFT, brazing with silver solder. The thermal fluid moves due to the density change of the cold and hot water. It is a mixture of ethyleneglycol-water and the heat is exchanged in the copper serpentine installed in the reservoir of 200 liters. Daily it warms 80 liters of water to 45ºC.

  18. CAPTURA DE CO2 MEDIANTE TRANSPORTADORES SÓLIDOS

    Directory of Open Access Journals (Sweden)

    Carmen Forero

    2011-01-01

    Full Text Available La evaluación de transportadores de oxígeno (TO, basados en CuO y NiO sobre Al2O3 y preparados por impregnación, se llevó a cabo en una planta piloto de dos lechos fluidizados interconectados de 500 Wte, donde se utilizaron tanto metano como gas de síntesis como gas combustible. Además, se estudió el efecto de diferentes impurezas presentes en el gas combustible como azufre o hidrocarburos ligeros en la eficacia de combustión del proceso y en el comportamiento de los TO. Los resultados obtenidos mostraron que ambos TO son adecuados para la captura de CO2 mediante transportadores sólidos de oxígeno en el proceso de combustión de metano, gas de síntesis o metano con impurezas como hidrocarburos ligeros o azufre en el gas.

  19. Detection of Coxiella burnetii in ticks by PCR and by PCR - Restriction Fragment Length Polymorphism (RFLP)

    International Nuclear Information System (INIS)

    Coxiella burnetii, as an obligata intracellular bacterium, is the etiologic agent of Q-fever. It is widely distributed in nature and is responsible for infection in various animals (cattle, sheep, goat) and humans. C. burnetii has been isolated from milk, ticks and human patients with acute and chronic Q fever. Ticks are the principal vectors and reservoirs of C. burnetii. Since over 40 species of ticks have been found to be infected with C. burnetii, ticks can serve as indicators of infection in nature. In this study, total of 2472 ticks (1446 female, 1021 male and 5 nymphs) were collected from 38 provinces of Turkey. The ticks were gathered into groups of 1 to 7 ticks as to the provinces, species and gender for DNA extraction. Following DNA extraction, the groups were examined for the presence of C. burtii by using the CB1and CB2. The ticks collected from the province of Denizli (56 in total) were gathered into 13 groups according to the species and gender. From these groups, 6 were positive for C. burnetii. The ticks collected from Ankara province, total of 160 ticks, were grouped into 53 as to their species and gender, only one group was found to be positive for C. burnetii. The specificities of PCR products were evaluated by restriction analysis. The positive PCR products were digested with the enzyme Taq1 and for bands in order of 118, 57, 43 and 39 bp's were appeared such as seen in the positive control DNA (C. burnetii Nine Mile RSA493)

  20. Animal Images and Metaphors in Animal Farm

    OpenAIRE

    Ping Sun

    2015-01-01

    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 ani...

  1. 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 sample...... results. Finally, the Microsporum PCR was positive for 10/10 guinea pig specimens from infected animals but for 0/2 of the control animal samples. The evaluation of the two PCR tests indicated excellent sensitivity and specificity.......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...

  2. 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.

  3. Validation of a Real Time PCR for Classical Swine Fever Diagnosis

    Directory of Open Access Journals (Sweden)

    Natanael Lamas Dias

    2014-01-01

    Full Text Available The viral disease classical swine fever (CSF, caused by a Pestivirus, is one of the major causes of economic losses for pig farming. The aim of this work was to validate a RT-qPCR using Taqman for detection of CSF in swine tissues. The parameters for the validation followed the specifications of the Manual of Diagnostic Tests and Vaccines for Terrestrial Animals of the World Organization for Animal Health (OIE and the guide ABNT NBR ISO/IEC 17025:2005. The analysis of the 5′NTR region of CSF virus was performed in 145 samples from 29 infected pigs and in 240 samples from 80 pigs originated in the Brazilian CSF-free zone. The tissues tested were spleen, kidney, blood, tonsils, and lymph nodes. Sequencing of the positive samples for 5′NTR region was performed to evaluate the specificity of the RT-qPCR. Tests performed for the RT-qPCR validation demonstrated that the PCR assay was efficient in detecting RNA from CSF virus in all materials from different tissues of infected animals. Furthermore, RNA from CSF virus was not detected in samples of swine originated from the Brazilian CSF-free zone. Hence, it is concluded that RT-qPCR can be used as a complementary diagnostic for CSF.

  4. Development and evaluation of PCR assay for detection of low levels of Cowdria ruminantium infection in Amblyomma ticks not detected by DNA probe.

    OpenAIRE

    Peter, T F; Deem, S L; Barbet, A F; Norval, R A; Simbi, B H; Kelly, P. J.; Mahan, S. M.

    1995-01-01

    The sensitivities of a PCR assay and a DNA probe assay were compared for the detection of Cowdria ruminantium in Amblyomma ticks that were fed on C. ruminantium-infected, clinically reacting, and recovered carrier animals. The PCR assay and DNA probe detected infection in 86.0 and 37.0%, respectively, of 100 ticks fed on a febrile animal. In 75 ticks fed on carrier animals, PCR and the DNA probe detected infection in 28.0 and 1.33% of ticks, respectively. This demonstrates that the DNA probe ...

  5. Animation of Antimicrobial Resistance

    Medline Plus

    Full Text Available ... En Español Search FDA Submit search Popular Content Home Food Drugs Medical Devices Radiation-Emitting Products Vaccines, ... Biologics Animal & Veterinary Cosmetics Tobacco Products Animal & Veterinary Home Animal & Veterinary Safety & Health Antimicrobial Resistance Animation of ...

  6. 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…

  7. 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....

  8. 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.

  9. Control mediante SCADA de un panel con elementos neumáticos

    OpenAIRE

    PERIS MARTINEZ, SERGI

    2015-01-01

    [ES] Se realiza y explica un control mediante un sistema SCADA sobre unos elementos neumáticos de agarre, posicionamiento y desplazamiento. Éstos estarán accionados con electroválvulas, controladas mediante un autómata programable. También se muestra lo que podría ser un ejemplo de un proceso industrial utilizando los elementos anteriores para hacer el control de dicho procedimiento.

  10. 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

  11. 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

  12. Carotenoids in Marine Animals

    OpenAIRE

    Takashi Maoka

    2011-01-01

    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 a...

  13. Estudio de las comunidades microbianas de embutidos fermentados ligeramente acidificados mediante técnicas moleculares. Estandarización, seguridad y mejora tecnológica.

    OpenAIRE

    Martín Juárez, Belén

    2005-01-01

    Los embutidos fermentados ligeramente acidificados son un grupo de productos tradicionales mediterráneos, caracterizados por un pH superior a 5,3.Para un control eficiente de la seguridad microbiológica de los embutidos se necesitan técnicas rápidas para la identificación y recuento de los microorganismos patógenos a estudiar. En el presente trabajo, se desarrolló una técnica para la enumeración de L. monocytogenes que combinó el método del número más probable y la identificación mediante PCR...

  14. 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.

  15. 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.

  16. Comparative analysis of conventional PCR and real-time PCR to diagnose shrimp WSD

    OpenAIRE

    Leal, C. A. G.; Carvalho-Castro, G.A.; Cottorello, A.C.; Leite, R. C.; Figueiredo, H. C. P.

    2013-01-01

    The aims of this study were to standard and optimize a qPCR protocol with FAM-BHQ1 probe, and to compare its sensitivity against TaqMan qPCR and PCR methods to diagnose shrimp WSD. The FAM-BHQ1 qPCR presented higher clinical sensitivity and showed to be a robust alternative to detect WSSV in clinical samples.

  17. 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.

  18. Detection of sulfonamide resistance genes via in situ PCR-FISH.

    Science.gov (United States)

    Gnida, Anna; Kunda, Katarzyna; Ziembińska, Aleksandra; Luczkiewicz, Aneta; Felis, Ewa; Surmacz-Górska, Joanna

    2014-01-01

    Due to the rising use of antibiotics and as a consequence of their concentration in the environment an increasing number of antibiotic resistant bacteria is observed. The phenomenon has a hazardous impact on human and animal life. Sulfamethoxazole is one of the sulfonamides commonly detected in surface waters and soil. The aim of the study was to detect sulfamethoxazole resistance genes in activated sludge biocenosis by use of in situ PCR and/or hybridization. So far no FISH probes for the detection of SMX resistance genes have been described in the literature. We have tested common PCR primers used for SMX resistance genes detection as FISH probes as well as a combination of in situ PCR and FISH. Despite the presence of SMX resistance genes in activated sludge confirmed via traditional PCR, the detection of the genes via microscopic visualization failed. PMID:25115110

  19. Application of Droplet Digital PCR to Validate Rift Valley Fever Vaccines.

    Science.gov (United States)

    Ly, Hoai J; Lokugamage, Nandadeva; Ikegami, Tetsuro

    2016-01-01

    Droplet Digital™ polymerase chain reaction (ddPCR™) is a promising technique that quantitates the absolute concentration of nucleic acids in a given sample. This technique utilizes water-in-oil emulsion technology, a system developed by Bio-Rad Laboratories that partitions a single sample into thousands of nanoliter-sized droplets and counts nucleic acid molecules encapsulated in each individual particle as one PCR reaction. This chapter discusses the applications and methodologies of ddPCR for development of Rift Valley fever (RVF) vaccine, using an example that measures RNA copy numbers of a live-attenuated MP-12 vaccine from virus stocks, infected cells, or animal blood. We also discuss how ddPCR detects a reversion mutant of MP-12 from virus stocks accurately. The use of ddPCR improves the quality control of live-attenuated vaccines in the seed lot systems. PMID:27076132

  20. 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…

  1. 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...

  2. 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 verify...

  3. 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...

  4. EVALUACIÓN DE LA ESTABILIDAD GENÉTICA MEDIANTE MARCADORES RAPD EN PLANTAS DE Ipomoea batatas

    Directory of Open Access Journals (Sweden)

    O. González

    2007-01-01

    mezcla homogénea de tejido de limbo foliar (1,0 g con nitrógeno líquido. La concentración de ADN se determinó por espectrofotometría y los materiales se evaluaron para 10 cebadores arbitrarios, según las recomendaciones de los protocolos de la firma comercial Operon Technologies, que fueron: OPF-15, OPF-14, OPA-13, OPF-13, OPF-04, OPF-07, OPF-01, OPF-03, OPA-12 y OPF-05. Los productos de la amplificación (PCR se separaron mediante electroforesis en gel de agarosa al 1.5 %, en solución amortiguadora TBE y se tiñeron con bromuro de etidio antes de ser visualizados en un transiluminador ultravioleta. La comparación de los patrones de amplificación obtenidos se realizó evaluando las bandas de forma binaria por su presencia (1 y ausencia (0 para cada uno de los tratamientos, donde se pudo observar un monomorfismo total en las bandas del material donante, en comparación con el procedente de vitroplantas de embriones somáticos.

  5. 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.

  6. Urinary PCR as an increasingly useful tool for an accurate diagnosis of leptospirosis in livestock.

    Science.gov (United States)

    Hamond, C; Martins, G; Loureiro, A P; Pestana, C; Lawson-Ferreira, R; Medeiros, M A; Lilenbaum, W

    2014-03-01

    The aim of the present study was to consider the wide usage of urinary PCR as an increasingly useful tool for an accurate diagnosis of leptospirosis in livestock. A total of 512 adult animals (300 cattle, 138 horses, 59 goats and 15 pigs), from herds/flocks with reproductive problems in Rio de Janeiro, Brazil was studied by serology and urinary PCR. From the 512 serum samples tested, 223 (43.5 %) were seroreactive (cattle: 45.6 %, horses: 41.3 %, goats: 34%and pigs: 60 %). PCR detected leptospiral DNA in 32.4 % (cattle: 21.6 %, horses: 36.2 %, goats: 77.4 % and pigs: 33.3 %. To our knowledge there is no another study including such a large number of samples (512) from different species, providing a comprehensive analysis of the usage of PCR for detecting leptospiral carriers in livestock. Serological and molecular results were discrepant, regardless the titre, what was an expected outcome. Nevertheless, it is impossible to establish agreement between these tests, since the two methodologies are conducted on different samples (MAT - serum; PCR - urine). Additionally, the MAT is an indirect method and PCR is a direct one. In conclusion, we have demonstrated that urinary PCR should be considered and encouraged as an increasingly useful tool for an accurate diagnosis of leptospirosis in livestock. PMID:24222053

  7. 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.

  8. 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.

  9. Genotyping of plant and animal samples without prior DNA purification.

    Science.gov (United States)

    Chum, Pak Y; Haimes, Josh D; André, Chas P; Kuusisto, Pia K; Kelley, Melissa L

    2012-01-01

    The Direct PCR approach facilitates PCR amplification directly from small amounts of unpurified samples, and is demonstrated here for several plant and animal tissues (Figure 1). Direct PCR is based on specially engineered Thermo Scientific Phusion and Phire DNA Polymerases, which include a double-stranded DNA binding domain that gives them unique properties such as high tolerance of inhibitors. PCR-based target DNA detection has numerous applications in plant research, including plant genotype analysis and verification of transgenes. PCR from plant tissues traditionally involves an initial DNA isolation step, which may require expensive or toxic reagents. The process is time consuming and increases the risk of cross contamination. Conversely, by using Thermo Scientific Phire Plant Direct PCR Kit the target DNA can be easily detected, without prior DNA extraction. In the model demonstrated here, an example of derived cleaved amplified polymorphic sequence analysis (dCAPS) is performed directly from Arabidopsis plant leaves. dCAPS genotyping assays can be used to identify single nucleotide polymorphisms (SNPs) by SNP allele-specific restriction endonuclease digestion. Some plant samples tend to be more challenging when using Direct PCR methods as they contain components that interfere with PCR, such as phenolic compounds. In these cases, an additional step to remove the compounds is traditionally required. Here, this problem is overcome by using a quick and easy dilution protocol followed by Direct PCR amplification (Figure 1). Fifteen year-old oak leaves are used as a model for challenging plants as the specimen contains high amounts of phenolic compounds including tannins. Gene transfer into mice is broadly used to study the roles of genes in development, physiology and human disease. The use of these animals requires screening for the presence of the transgene, usually with PCR. Traditionally, this involves a time consuming DNA isolation step, during which DNA

  10. Assessment of the real-time PCR and different digital PCR platforms for DNA quantification.

    Science.gov (United States)

    Pavšič, Jernej; Žel, Jana; Milavec, Mojca

    2016-01-01

    Digital PCR (dPCR) is beginning to supersede real-time PCR (qPCR) for quantification of nucleic acids in many different applications. Several analytical properties of the two most commonly used dPCR platforms, namely the QX100 system (Bio-Rad) and the 12.765 array of the Biomark system (Fluidigm), have already been evaluated and compared with those of qPCR. However, to the best of our knowledge, direct comparison between the three of these platforms using the same DNA material has not been done, and the 37 K array on the Biomark system has also not been evaluated in terms of linearity, analytical sensitivity and limit of quantification. Here, a first assessment of qPCR, the QX100 system and both arrays of the Biomark system was performed with plasmid and genomic DNA from human cytomegalovirus. With use of PCR components that alter the efficiency of qPCR, each dPCR platform demonstrated consistent copy-number estimations, which indicates the high resilience of dPCR. Two approaches, one considering the total reaction volume and the other considering the effective reaction size, were used to assess linearity, analytical sensitivity and variability. When the total reaction volume was considered, the best performance was observed with qPCR, followed by the QX100 system and the Biomark system. In contrast, when the effective reaction size was considered, all three platforms showed almost equal limits of detection and variability. Although dPCR might not always be more appropriate than qPCR for quantification of low copy numbers, dPCR is a suitable method for robust and reproducible quantification of viral DNA, and a promising technology for the higher-order reference measurement method. PMID:26521179

  11. Source identification of airborne Escherichia coli of swine house surroundings using ERIC-PCR and REP-PCR.

    Science.gov (United States)

    Duan, Huiyong; Chai, Tongjie; Liu, Jianzhu; Zhang, Xingxiao; Qi, Chunhua; Gao, Jing; Wang, Yaling; Cai, Yumei; Miao, Zengmin; Yao, Meiling; Schlenker, Gerd

    2009-07-01

    Evidence is mounting that microorganisms originating from livestock impact the air quality of the animal houses themselves and the public in the surrounding neighborhoods. The aim of this study was to develop efficient bacterial source tracking capabilities to identify sources of Escherichia coli aerosol pollution caused by pigs. Airborne E. coli were isolated from indoor air, upwind air (10 and 50 m away) and downwind air samples (10, 50, 100, 200 and 400 m away) for five swine houses using six-stage Andersen microbial samplers and Reuter-Centrifugal samplers (RCS). E. coli strains from pig fecal samples were also collected simultaneously. The enterobacterial repetitive intergenic consensus polymerize chain reaction (ERIC-PCR) and the repetitive extragenic palindromic (REP-PCR) approaches were used to study the genetic variability and to determine the strain relationships among E. coli isolated from different sites in each swine house. Results showed that 35.1% (20/57) of the bacterial DNA fingerprints from the fecal isolates matched with the corresponding strains isolated from indoor and downwind air samples (similarity > or = 90%). E. coli strains from the indoor and downwind air samples were closely related to the E. coli strains isolated from feces, while those isolated from upwind air samples (swine house C) had low similarity (61-69%). Our results suggest that some strains isolated from downwind and indoor air originated in the swine feces. Effective hygienic measures should be taken in animal farms to prevent or minimize the downwind spread of microorganism aerosol. PMID:19349045

  12. 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.

  13. Detection of classical swine fever virus (CSFV) in clinical samples by RT-PCR assay in clinical samples by RT-PCR assay using different pairs of primers

    International Nuclear Information System (INIS)

    The aim was to compare the efficiency of RT-PCT assays using four pairs of primers selected from different regions of the CSFV genome for the detection of CSFV in clinical samples of swine and wild boars. The four RT-PCR assays were able to detect CSFV in all 20 clinical samples which had been collected from dead swine and wild boars during the outbreaks of CSF in Slovakia in 1993 and 1994. The quality of the selected RT-PCR primers was determined as follows: gp55L/gp55U (E2), 324/326 (5'-NC), S1/S2 (NS5B) and gp54L/gp54U (NS2 genomic region). We conclude that gp55L/gp55U primers are the most suitable for direct detection of CSFV by RT-PCR in tissue homogenates of diseased animals

  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. Morris Animal Foundation

    Science.gov (United States)

    ... the transmission of serious illnesses. Read more » Morris Animal Foundation Receives $750,000 Grant for Cancer Studies. ... Give Partners Become a Partner Meet Our Partners Animal Lovers Our Work Ways to Give Pet Health ...

  16. "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.

  17. Animals in Education.

    Science.gov (United States)

    Rowan, Andrew N.

    1981-01-01

    Summarizes viewpoints on the use of animals in science experiments in the biology classroom, including those of teachers, education researchers, biomedical scientists, science education administrators, and animal welfare advocates. (Author/CS)

  18. Animation of Antimicrobial Resistance

    Medline Plus

    Full Text Available ... FDA Submit search Popular Content Home Food Drugs Medical Devices Radiation-Emitting Products Vaccines, Blood & Biologics Animal & ... by Product Area Product Areas back Food Drugs Medical Devices Radiation-Emitting Products Vaccines, Blood & Biologics Animal & ...

  19. Interaction between animal personality and animal cognition

    OpenAIRE

    Claudio CARERE, Charles LOCURTO

    2011-01-01

    The study of animal personality has attracted considerable attention, as it has revealed a number of similarities in personality between humans and several nonhuman species. At the same time the adaptive value and evolutionary maintenance of different personalities are the subject of debate. Since Pavlov’s work on dogs, students of comparative cognition have been aware that animals display vast individual differences on cognitive tasks, and that these differences may not be entirely accounted...

  20. PcrA function in plasmid replication

    OpenAIRE

    Chisty, L. T.

    2014-01-01

    PcrA is a DNA helicase involved in unwinding plasmi ds as a part of a complex in asymmetric rolling - circle replication of certain plasmids carrying antibiotic resistance genes. PcrA translocates on single stranded DNA by coupling ATP hydrolysis to movement on DNA. Initiator protein, RepD is required to nick supercoiled plasmid site - specifically and open an ssDNA stretch that PcrA can bind. The presence of RepD is needed throughout plasmid unwinding to maintain processivity. Using fluo...

  1. Generic Face Animation

    OpenAIRE

    Cerda, Mauricio; Valenzuela, Renato; Hitschfeld-Kahler, Nancy; Terissi, Lucas; Gomez, Juan C.

    2010-01-01

    International audience In computer vision, the animation of objects has attracted a lot attention, specially the animations of 3D face models. The animation of face models requires in general to manually adapt each generic movement (open/close mouth) to each specific head geometry. In this work we propose a technique for the animation of any face model avoiding most of the manual intervention. In order to achieve this we assume that: (1) faces, despite obvious differences are quite similar...

  2. Imaging of Awake Animals

    OpenAIRE

    Wilkinson, Thomas

    2015-01-01

    The 3Rs of reduction, refinement and replacement are the guiding principles of animal research and embedded in national and international legislation regulating the use of animals in scientific procedures. Awake imaging by MRI of rodents can offer a reduction by increasing the quality of scientific data through longitudinal imaging using less animals by avoiding a serial sacrifice design and refinement through reducing the stressful effects animals are exposed to, in comparison to existing mo...

  3. Biopolitics: Animals, meat, food

    OpenAIRE

    Janović Nikola

    2009-01-01

    The general idea of this text is to reflect biopolitical constitution of the society and its implications related to the issues of animal welfare. Since animal in biopolitical formation is technically reduced to an object - commodity for contentment of the industry and of the people needs - critical public advisories are calling from moral, ethical and legal standpoint for attention to the fact that is necessary to protect animals from the unnecessary exploitation. It is obvious that animal p...

  4. 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

  5. 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.

  6. Activities of the Animal Production and Health Laboratory (Animal Production and Health Newsletter, No. 60, July 2014)

    International Nuclear Information System (INIS)

    This article provides information on: Genetic variation on the control of resistance to infectious diseases in small ruminants for improving animal productivity; Genetic characterization of indigenous livestock breeds; Testing irradiation technology for potential use in trypanosome vaccine development; Strengthening animal disease diagnostic capacities in veterinary laboratories in sub-Saharan Africa; Proficiency testing for Peste des Petits Ruminants (PPR) diagnosis by Nucleic Acid Amplification (RT-PCR). Information on Fellows is also provided

  7. Comparison of Galactomannan Detection, PCR-Enzyme-Linked Immunosorbent Assay, and Real-Time PCR for Diagnosis of Invasive Aspergillosis in a Neutropenic Rat Model and Effect of Caspofungin Acetate

    OpenAIRE

    Scotter, Jennifer M.; Chambers, Stephen T

    2005-01-01

    The performance of different in vitro diagnostic tests for the diagnosis of invasive aspergillosis (IA) was investigated in a transiently neutropenic rat model. Rats were immunosuppressed with cyclophosphamide and then inoculated intravenously with 1.5 × 104 CFU Aspergillus fumigatus spores. Animals were then either treated with caspofungin acetate, 1 mg/kg/day for 7 days, or not treated. PCR-enzyme-linked immunosorbent assay (ELISA), real-time PCR, and galactomannan (GM) detection were perfo...

  8. Bioethics in animal experimentation

    OpenAIRE

    Popa V.I.; Lascar I.; Valcu M.; Sebe Ioana Teona; Caraban B.; Margina Arina Cristiana

    2015-01-01

    Animal experiments are used on a large scale worldwide in order to develop or to refine new medicines, medicinal products or surgical procedures. It is morally wrong to cause animals to suffer, this is why animal experimentation causes serious moral problems.

  9. Animation of Antimicrobial Resistance

    Medline Plus

    Full Text Available ... Translation - Animation of Antimicrobial Resistance (WMV - 19.2MB) Chinese Translation - Animation of Antimicrobial Resistance (WMV - 19.2MB) ... by Product Area Product Areas back Food Drugs Medical Devices Radiation-Emitting Products Vaccines, Blood & Biologics Animal & ...

  10. Animal Models for imaging

    OpenAIRE

    Croft, Barbara Y.

    2002-01-01

    Animal models can be used in the study of disease. This chapter discusses imaging animal models to elucidate the process of human disease. The mouse is used as the primary model. Though this choice simplifies many research choices, it necessitates compromises for in vivo imaging. In the future, we can expect improvements in both animal models and imaging techniques.

  11. 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.

  12. Industralization of Animal Agriculture

    OpenAIRE

    Oya S. Erdogdu; David Hennessy

    2003-01-01

    The economic concerns and the technological developments increased control over nature and nurture in the animal agriculture. That changed the seasonality pattern of the supply side and lead to structural change in the animal agriculture together with the demand side factors. In this study we focused on the supply side factors and document the ‘industralization’ of the animal agricultural production.

  13. Bioethics in animal experimentation

    Directory of Open Access Journals (Sweden)

    Popa V.I.

    2015-11-01

    Full Text Available Animal experiments are used on a large scale worldwide in order to develop or to refine new medicines, medicinal products or surgical procedures. It is morally wrong to cause animals to suffer, this is why animal experimentation causes serious moral problems.

  14. 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.

  15. 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.

  16. Comparative Analysis of Cultural Isolation and Pcr Based Assay for Detection of Campylobacter Jejuni In Food and Faecal Samples

    OpenAIRE

    Singh, Harkanwaldeep; Rathore, R. S.; Singh, Satparkash; Cheema, Pawanjit Singh

    2011-01-01

    In the present study, the efficacy of polymerase chain reaction (PCR) based on mapA gene of C. jejuni was tested for detection of Campylobacter jejuni in naturally infected as well as spiked faecal and food samples of human and animal origin. Simultaneously, all the samples were subjected to the cultural isolation of organism and biochemical characterization. The positive samples resulted in the amplification of a DNA fragment of size ~589 bp in PCR assay whereas the absence of such amplicon ...

  17. Comparative analysis of cultural isolation and PCR based assay for detection of Campylobacter jejuni in food and faecal samples

    OpenAIRE

    Harkanwaldeep Singh; Rathore, R. S.; Satparkash Singh; Pawanjit Singh Cheema

    2011-01-01

    In the present study, the efficacy of polymerase chain reaction (PCR) based on mapA gene of C. jejuni was tested for detection of Campylobacter jejuni in naturally infected as well as spiked faecal and food samples of human and animal origin. Simultaneously, all the samples were subjected to the cultural isolation of organism and biochemical characterization. The positive samples resulted in the amplification of a DNA fragment of size ~589 bp in PCR assay whereas the absence of such amplicon ...

  18. Animal models of dementia

    DEFF Research Database (Denmark)

    Olsson, I. Anna S.; Sandøe, Peter

    2011-01-01

    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......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...... bears responsibility for the use of animals in disease models?...

  19. Expression-PCR: A rapid method for in vitro expression of PCR products

    International Nuclear Information System (INIS)

    We present a rapid and simple method called Expression-PCR (E-PCR) for in vitro synthesis of protein from genomic, plasmid or reverse transcribed DNA. Expression-PCR is a procedure for installing transcription and translation signals to genes of interest allowing their efficient expression in vitro. These signals are contained in an in vitro expression cassette (EC) containing an untranslated leader sequence from alfalfa mosaic virus (AMV-UTL) directly downstream form the T7 bacteriophage promoter site. When this EC is spliced to a PCR product, it produces a suitable template for direct in vitro transcription and translation. This methodology permits the rapid analysis of gene products without the need for cloning or in vivo expression. E-PCR represents a significant improvement over current in vitro expression systems, most notably in time-savings, versatility of gene expression and compatibility with rapid PCR-based site-directed mutagenesis procedures. (author). 20 refs, 2 figs

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

    International Nuclear Information System (INIS)

    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 32P 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)

  1. 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)

  2. Hot Start PCR with heat-activatable primers: a novel approach for improved PCR performance

    OpenAIRE

    Lebedev, Alexandre V.; Paul, Natasha; Yee, Joyclyn; Timoshchuk, Victor A.; Shum, Jonathan; Miyagi, Kei; Kellum, Jack; Hogrefe, Richard I.; Zon, Gerald

    2008-01-01

    The polymerase chain reaction (PCR) is widely used for applications which require a high level of specificity and reliability, such as genetic testing, clinical diagnostics, blood screening, forensics and biodefense. Great improvements to PCR performance have been achieved by the use of Hot Start activation strategies that aim to prevent DNA polymerase extension until more stringent, higher temperatures are reached. Herein we present a novel Hot Start activation approach in PCR where primers ...

  3. Viral diagnostics in the era of digital PCR

    OpenAIRE

    Sedlak, Ruth Hall; Jerome, Keith R.

    2012-01-01

    Unlike quantitative PCR (qPCR), digital PCR (dPCR) achieves sensitive and accurate absolute quantitation of a DNA sample without the need for a standard curve. A single PCR reaction is divided into many separate reactions that each have a positive or negative signal. By applying Poisson statistics, the number of DNA molecules in the original sample is directly calculated from the number of positive and negative reactions. The recent availability of multiple commercial dPCR platforms has led t...

  4. Multiplex allele-specific target amplification based on PCR suppression

    OpenAIRE

    Broude, Natalia E.; Zhang, Lingang; Woodward, Karen; Englert, David; Cantor, Charles R.

    2001-01-01

    We have developed a strategy for multiplex PCR based on PCR suppression. PCR suppression allows DNA target amplification with only one sequence-specific primer per target and a second primer that is common for all targets. Therefore, an n-plex PCR would require only n + 1 primers. We have demonstrated uniform, efficient amplification of targeted sequences in 14-plex PCR. The high specificity of suppression PCR also provides multiplexed amplification with allele specifi...

  5. 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. PMID:27393656

  6. 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.

  7. Tratamiento de las fracturas diafisarias de húmero mediante osteosíntesis con placa

    OpenAIRE

    Zamora Rodríguez, J. M.; Modrego Aranda, Francisco Javier; Seral García, Belén; Seral Iñigo, Fernando

    2002-01-01

    Hemos tratado 22 facturas agudas diafisarias de húmero mediante reducción abierta y fijación con placa AO entre 1991 y 1999. Todas las fracturas excepto una consolidaron en un plazo medio de 94 días. De acuerdo con el criterio de Brumback los resultados fueron excelentes o buenos en el 90% de los casos. La complicación postoperativa más importante fue la parálisis del nervio radial en 3 casos; todos se recuperaron espontáneamente en un plazo medio de 108 días. La fijación interna mediante pla...

  8. Reconocimiento de textos antiguos mediante técnicas de visión artficial

    OpenAIRE

    Rodríguez Vallejo, Moisés

    2013-01-01

    El objetivo de este proyecto es reconocer caracteres de textos antiguos mediante t ecnicas de visi on arti cial y redes neuronales. Distinguimos dos fases en el proceso: extracci on de los caracteres de las im agenes de los textos antiguos; y el reconocimiento mediante redes neuronales. En la primera utilizaremos las librer as de OpenCV para conseguir el objetivo, mientras que en la segunda fase partimos de un c odigo de una red neuronal de 3 capas de la Universidad Carnegie M...

  9. Simulación y llenado de moldes mediante Moldflow®

    OpenAIRE

    Peidró Roy, Alejandro

    2015-01-01

    El objetivo del presente proyecto es recopilar los conocimientos necesarios para que los alumnos de Formación Professional del ciclo formativo de Diseño mecánico en el módulo M5 Moldes poliméricos, partiendo de cero, puedan diseñar un pieza de plástico y su molde de inyección asociado mediante el software Catia® en los módulos de Part desing, Generative Shape desing y Mold Tooling desing, simularla mediante Moldflow® Plastics, rediseñarla y optimizarla, de forma que el documento que se redact...

  10. Small Animal Retinal Imaging

    Science.gov (United States)

    Choi, WooJhon; Drexler, Wolfgang; Fujimoto, James G.

    Developing and validating new techniques and methods for small animal imaging is an important research area because there are many small animal models of retinal diseases such as diabetic retinopathy, age-related macular degeneration, and glaucoma [1-6]. Because the retina is a multilayered structure with distinct abnormalities occurring in different intraretinal layers at different stages of disease progression, there is a need for imaging techniques that enable visualization of these layers individually at different time points. Although postmortem histology and ultrastructural analysis can be performed for investigating microscopic changes in the retina in small animal models, this requires sacrificing animals, which makes repeated assessment of the same animal at different time points impossible and increases the number of animals required. Furthermore, some retinal processes such as neurovascular coupling cannot be fully characterized postmortem.

  11. 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 disgust operate and defend their conceptual and theoretical coherence against common objections. I also outline an explanatory challenge for disgust researchers. Both types of disgust indicate that a wide variety of animals produce aversive and avoidant reactions in human beings. This seems somewhat odd......, given the prominence of animals in human lives. The challenge, then, is explaining how humans cope with the presence of animals. I propose, as a hypothesis for further exploration, that we cope with animals, and our disgust responses to them, by attributing mental states that mark them as inferior...

  12. Animal models of asthma

    OpenAIRE

    Bates, Jason H.T.; Rincon, Mercedes; Irvin, Charles G.

    2009-01-01

    Studies in animal models form the basis for much of our current understanding of the pathophysiology of asthma, and are central to the preclinical development of drug therapies. No animal model completely recapitulates all features of the human disease, however. Research has focused primarily on ways to generate allergic inflammation by sensitizing and challenging animals with a variety of foreign proteins, leading to an increased understanding of the immunological factors that mediate the in...

  13. Animal Violence Demystified

    OpenAIRE

    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 (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 (char...

  14. Animal Model of Dermatophytosis

    OpenAIRE

    Tsuyoshi Shimamura; Nobuo Kubota; Kazutoshi Shibuya

    2012-01-01

    Dermatophytosis is superficial fungal infection caused by dermatophytes that invade the keratinized tissue of humans and animals. Lesions from dermatophytosis exhibit an inflammatory reaction induced to eliminate the invading fungi by using the host’s normal immune function. Many scientists have attempted to establish an experimental animal model to elucidate the pathogenesis of human dermatophytosis and evaluate drug efficacy. However, current animal models have several issues. In the presen...

  15. PRINCIPLES OF ANIMAL BREEDING

    OpenAIRE

    Sonja Jovanovac

    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 ...

  16. Are ticks venomous animals?

    OpenAIRE

    Cabezas-Cruz, Alejandro; James J Valdés

    2014-01-01

    Introduction As an ecological adaptation venoms have evolved independently in several species of Metazoa. As haematophagous arthropods ticks are mainly considered as ectoparasites due to directly feeding on the skin of animal hosts. Ticks are of major importance since they serve as vectors for several diseases affecting humans and livestock animals. Ticks are rarely considered as venomous animals despite that tick saliva contains several protein families present in venomous taxa and that many...

  17. The representative animal

    OpenAIRE

    Harrison, J. M.

    1994-01-01

    The anthropocentric approach to the study of animal behavior uses representative nonhuman animals to understand human behavior. This approach raises problems concerning the comparison of the behavior of two different species. The datum of behavior analysis is the behavior of humans and representative animal phenotypes. The behavioral phenotype is the product of the ontogeny and phylogeny of each species, and this requires that contributions of genotype as well as behavioral history to experim...

  18. Animal Production in Turkey

    OpenAIRE

    SARICA, Şenay; Ulutaş, Zafer; ŞAHİN, Aziz

    2004-01-01

    Animal sector in Turkey has changed considerably in the last few years. Although the most significant advancements have occurred in the poultry sector, the cattle and small ruminants sector could not achieve similar improvements. Reasons of the depression in the cattle and small ruminants sector are the lack of breeding animal materials and high quality feed sources, insufficient disease control, disorganized and small size of the animal farms, lack of infrastructure, poor education levels of...

  19. Thinking with animals

    OpenAIRE

    2015-01-01

    they also enlist them to symbolize, dramatize, and illuminate aspects of humans' experience and fantasy. Humans merge with animals in stories, films, philosophical speculations, and scientific treatises. In their performance on many stages and in different ways, animals move us to think." "Essays in the book investigate the changing patterns of anthropomorphism across different time periods and settings, as well as their transformative effects, both figuratively and literally, upon animals, h...

  20. 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

  1. The dying animal.

    Science.gov (United States)

    Pierce, Jessica

    2013-12-01

    The study of animal death is poised to blossom into an exciting new interdisciplinary field-and one with profound relevance for bioethics. Areas of interest include the biology and evolution of death-related behavior in nonhuman animals, as well as human social, psychological, cultural, and moral attitudes toward and practices related to animal death. In this paper, I offer a brief overview of what we know about death-related behavior in animals. I will then sketch some of the bioethical implications of this emerging field of research. PMID:24092402

  2. Animal-free toxicology

    DEFF Research Database (Denmark)

    Knudsen, Lisbeth E

    2013-01-01

    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....

  3. Diagnosis of animal allergy.

    Science.gov (United States)

    Patterson, R

    1987-01-01

    The aims of the diagnostic evaluation are to establish the presence and severity of disease and the importance of animal exposure as the etiology of the disease. The evaluation of the importance of animals may be part of a general allergy evaluation or specifically directed toward an animal in certain cases, such as occupational exposure. The diagnostic techniques are medical history, physical examination, allergy skin tests or in vitro tests for IgE antibody and correlation of improvement in symptoms with animal avoidance. PMID:3477684

  4. Political Communication with Animals

    OpenAIRE

    Meijer, E

    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 Donaldson and Will Kymlicka present in Zoopolis (2011). I will argue their political theory is an important step forward in the debate about animal rights, because it proposes to see animals as po...

  5. Diagnostic PCR of dermatophytes--an overview.

    Science.gov (United States)

    Gräser, Yvonne; Czaika, Viktor; Ohst, Torsten

    2012-10-01

    The prevalence of onychomycosis is increasing steadily, sevenfold alone in the US within the last twenty years. An important aspect in this development is the demographic development of the human population of the industrial countries like Germany. A fast and accurate laboratory diagnosis is essential for successful treatment because 50% of the cases are misdiagnosed when relying on the clinical appearance only. The current diagnosis of dermatophytosis, based on direct microscopy and culture of the clinical specimen, is problematic given the lacking specificity of the former and the length of time needed for the latter. Molecular techniques can help to solve these problems. In recent years, a number of in-house PCR assays have been developed to identify dermatophytes directly from clinical specimens. Based on the "Mikrobiologisch-infektiologischen Qualitätsstandards (MIQ) für Nukleinsäure-Amplifikationstechniken" and the MIQE guideline (Minimum Information for Publication of Quantitative Real-Time PCR Experiments) 11 studies are reviewed which were published between 2007 and 2010. The present article evaluates the quality of the PCR assays regarding false positive and false negative results due to contamination, PCR format, statistical analysis, and diagnostic performance of the studies. It shows that we are only at the beginning of providing high quality PCR diagnosis of dermatophytes. PMID:23013298

  6. 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.

  7. Recycling technology of sugar industry by-products for animal feeding

    OpenAIRE

    Yadira Suárez Rodríguez; Arael Martínez Teruel; Luis B. Ramos Sánchez; María Caridad Julián

    2006-01-01

    En este trabajo se presenta el desarrollo de una tecnología de reciclaje y enriquecimiento proteico mediante fermentación en estado sólido de los subproductos de la industria azucarera para su posterior utilización como alimento animal. A partir de un estudio bibliográfico sobre los aspectos más importantes de las tecnologías actuales de fabricación de alimentos para el consumo animal y las herramientas para el desarrollo de tecnologías de fermentaci ón en medios sólidos se ha desarrollado un...

  8. 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,

  9. 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.

  10. 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 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.

  11. 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…

  12. Animation of Antimicrobial Resistance

    Medline Plus

    Full Text Available ... FDA Submit search Popular Content Home Food Drugs Medical Devices Radiation-Emitting Products Vaccines, Blood & Biologics Animal & Veterinary ... by Product Area Product Areas back Food Drugs Medical Devices Radiation-Emitting Products Vaccines, Blood & Biologics Animal & Veterinary ...

  13. 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…

  14. 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

  15. Animation of Antimicrobial Resistance

    Medline Plus

    Full Text Available ... Translation - Animation of Antimicrobial Resistance (WMV - 19.2MB) Chinese Translation - Animation of Antimicrobial Resistance (WMV - 19.2MB) ... FEAR Act Site Map Transparency Website Policies U.S. Food and Drug Administration 10903 New Hampshire Avenue Silver ...

  16. Endangered Animals. Second Grade.

    Science.gov (United States)

    Popp, Marcia

    This second grade teaching unit centers on endangered animal species around the world. Questions addressed are: What is an endangered species? Why do animals become extinct? How do I feel about the problem? and What can I do? Students study the definition of endangered species and investigate whether it is a natural process. They explore topics…

  17. First Aid: Animal Bites

    Science.gov (United States)

    ... Story" 5 Things to Know About Zika & Pregnancy First Aid: Animal Bites KidsHealth > For Parents > First Aid: Animal Bites Print A A A Text Size ... For Kids For Parents MORE ON THIS TOPIC First Aid & Safety Center Infections That Pets Carry Dealing With ...

  18. Ode to an Animal

    Science.gov (United States)

    Nelken, Miranda

    2008-01-01

    People know little about the non-domesticated animals that live around them. Somehow, they seem remote. In stories they hear about them, animals are often acting, speaking, and dressing like people. This article presents a lesson where students learn about the native species of their area while exploring the concept of interdependence through…

  19. Reduction of heteroduplex formation in PCR amplification

    Czech Academy of Sciences Publication Activity Database

    Michu, Elleni; Mráčková, Martina; Vyskot, Boris; Žlůvová, Jitka

    2010-01-01

    Roč. 54, č. 1 (2010), s. 173-176. ISSN 0006-3134 R&D Projects: GA AV ČR(CZ) KJB600040801; GA ČR(CZ) GD204/09/H002; GA AV ČR(CZ) IAA600040801; GA MŠk(CZ) LC06004 Institutional research plan: CEZ:AV0Z50040507; CEZ:AV0Z50040702 Keywords : polymerase chain reaction * reconditioning PCR * mixed-template PCR Subject RIV: BO - Biophysics Impact factor: 1.582, year: 2010

  20. 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.

  1. 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

    2015-06-01

    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

  2. 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 week......, 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 of...

  3. Interaction between animal personality and animal cognition

    Directory of Open Access Journals (Sweden)

    Claudio CARERE, Charles LOCURTO

    2011-08-01

    Full Text Available The study of animal personality has attracted considerable attention, as it has revealed a number of similarities in personality between humans and several nonhuman species. At the same time the adaptive value and evolutionary maintenance of different personalities are the subject of debate. Since Pavlov’s work on dogs, students of comparative cognition have been aware that animals display vast individual differences on cognitive tasks, and that these differences may not be entirely accounted for differences in cognitive abilities. Here, we argue that personality is an important source of variation that may affect cognitive performance and we hypothesise mutual influences between personality and cognition across an individual’s lifespan. In particular, we suggest that: 1 personality profiles may be markers of different cognitive styles; 2 success or failure in cognitive tasks could affect different personalities differently; 3 ontogenetic changes of personality profiles could be reflected in changes in cognitive performance. The study of such interplay has implications in animal welfare as well as in neuroscience and in translational medicine [Current Zoology 57 (4: 491–498, 2011].

  4. Animal Health in Albania

    International Nuclear Information System (INIS)

    The animal health service policy in Albania represents an integral component of overall governmental, social and economic policy in the field of agricultural and rural development, public health, food processing and import/export of animal products. In order to obtain the necessary political, economic and public support, the animal health service attempts to contribute effectively to the overall development of the country which aims at improving the standards of living of its inhabitants. Practical means of contributing to national development include reducing food loses due to animal morbidity and mortality, increasing the productivity of the livestock population, protecting human health against zoonotic diseases and ensuring humane treatment of animals. An animal health strategy contributes to the creation of conditions necessary for uninterrupted animal disease surveillance and control in the country. The main animal health problem in Albania is brucellosis in ruminants, caused by B. melitensis. This infection currently affects the entire country, reaching a prevalence of 10% in several districts. The latest and most severe outbreaks of classical swine fever were identified on 1996 when 5 515 animals were infected and 3 683 animals died. The circulation of bluetongue virus (BTV) was detected for the first time in Albania in 2002 with a seroprevalence of 15%. The evidence of BTV circulation in Albania and the absence of the main vector C. imicola suggest that other Culicoides species could be implicated in virus transmission. H5N1 avian influenza in Albania was confirmed in March 2006 in backyard flocks in the villages of Cuke and Peze-Helmes. In both villages there were no human cases. Rabies was of concern in Albania from 1928 until 1976. The disease re-emerged in March 2001 in the village of Morine in Kukes district affecting a domestic dog and three persons were bitten. Other cases have been reported in northern Albania. (author)

  5. ANALYSIS OF POLYMORPHISM OF ALPHA S1 CASEIN OF SLOVAK PINZGAU CATTLE BY PCR-RFLP

    Directory of Open Access Journals (Sweden)

    MARTINA MILUCHOVÁ

    2013-07-01

    Full Text Available The work was oriented to identification of -s1 casein gene polymorphism and analysis of genotype structure in population of Slovak Pinzgau cattle. The material involved 93 cattle. Bovine genomic DNA was isolated by fenol-chlorophorm deprotenization and ethanol precipitation and used in order to estimate -s1 casein genotypes by means of PCR-RFLP method. The PCR products were digested with MaeIII restriction enzyme. In the population included in the study there were homozygote genotype BB (81 animals and heterozygote genotype BC (12 animals. Homozygote genotype CC has not been observed. In the total population of cattle homozygotes BB – 0.871 were the most frequent, while BC – 0.129 were the least frequent ones. This suggests a superiority of allele B – 0.9355.

  6. 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.

  7. 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

  8. Determination of the presence of Babesia species in blood samples of cattle, camel and sheep in Iran by PCR

    OpenAIRE

    Khamesipour Faham; Doosti Abbas; Koohi Arman; Chehelgerdi Mohammad; Mokhtari-Farsani Abbas; Chengula Augustino Alfred

    2015-01-01

    Babesia species are protozoan parasites that parasitize the erythrocytes of domestic animals and humans, causing anemia in the host. The parasites cause a zoonotic disease known as babesiosis. Polymerase chain reaction (PCR) has proven to be very sensitive for detecting Babesia in blood samples of affected animals, particular in ruminants. The purpose of the current study was to determine the presence of Babesia spp. in blood samples obtained from 2 cattle,...

  9. Leptospira spp detection by Polymerase Chain Reaction (PCR) in clinical samples of captive black-capped Capuchin monkey (Cebus apella)

    OpenAIRE

    Scarcelli Eliana; Piatti Rosa Maria; Fedullo José Daniel Luzes; Simon Faiçal; Cardoso Maristela Vasconcellos; Castro Vanessa; Miyashiro Simone; Genovez Margareth Élide

    2003-01-01

    Leptospirosis is a widely distributed zoonosis that affects domestic and wild animals, and that has the man as the end point of its epidemiological chain. Leptospirosis diagnosis in primates is more difficult than in other animal species, as clinical signs and lesions are less evident and antibody response is detected only for short periods. The aim of this article was to describe the detection of Leptospira spp using polymerase chain reaction (PCR), in clinical samples from one captive black...

  10. Validation of a Real Time PCR for Classical Swine Fever Diagnosis

    OpenAIRE

    Natanael Lamas Dias; Antônio Augusto Fonseca Júnior; Anapolino Macedo de Oliveira; Érica Bravo Sales; Bruna Rios Coelho Alves; Fernanda Alves Dorella; Marcelo Fernandes Camargos

    2014-01-01

    The viral disease classical swine fever (CSF), caused by a Pestivirus, is one of the major causes of economic losses for pig farming. The aim of this work was to validate a RT-qPCR using Taqman for detection of CSF in swine tissues. The parameters for the validation followed the specifications of the Manual of Diagnostic Tests and Vaccines for Terrestrial Animals of the World Organization for Animal Health (OIE) and the guide ABNT NBR ISO/IEC 17025:2005. The analysis of the 5′NTR region of CS...

  11. Toxinotyping of Clostridium perfringens isolates by ELISA and PCR from lambs suspected of enterotoxemia

    OpenAIRE

    HADİMLİ*, Hasan Hüseyin; ERGANİŞ, Osman; SAYIN, Zafer; ARAS, Zeki

    2012-01-01

    Clostridium perfringens is a gram-positive, anaerobic bacterium that causes a wide range of diseases in humans and animals. The purposes of this study were to determine, using the enzyme-linked immunosorbent assay (ELISA), the clostridial toxins in intestinal samples of lambs with suspected enterotoxemia; to conduct molecular typing of C. perfringens isolates; and to investigate the presence of the genes of 4 major toxins (a, b, e, and i) in the isolates by polymerase chain reaction (PCR). Ac...

  12. 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...

  13. PCR and antibody methods: Research compares two cattle feed tests that detect bovine byproduct contaminants

    OpenAIRE

    Sawyer, Mary M.; Smith, Wayne L.; Rensen, Gabriel J.; Osburn, Bennie I.; Cullor, James S.

    2005-01-01

    Preventing the spread of mad cow disease through contaminated cattle feed is a major concern of beef and dairy producers, regulators and consumers around the world. Routine testing of cattle feeds for the presence of banned substances is a critical control point in assuring animal health and food safety. We compared the results of two test procedures (a real-time polymerase chain reaction [PCR] assay and a commercially available ruminant antibody detection kit) on five cattle rations spiked w...

  14. False-Positive Results after Environmental Pinworm PCR Testing due to Rhabditid Nematodes in Corncob Bedding

    OpenAIRE

    Leblanc, Mathias; Berry, Kristina; Graciano, Sandy; Becker, Brandon; Reuter, Jon D.

    2014-01-01

    Modern rodent colonies are housed in individually ventilated cages to protect the animals from contamination with adventitious pathogens. Standard health monitoring through soiled-bedding sentinels does not always detect infections, especially in the context of low pathogen prevalence. Recently proposed alternatives include analyzing environmental samples from the cages or rack exhaust by PCR to improve the detection of rodent pathogens but optimal sampling strategies have not yet been establ...

  15. Multiplex PCR for Differential Identification of Broad Tapeworms (Cestoda: Diphyllobothrium) Infecting Humans

    Czech Academy of Sciences Publication Activity Database

    Wicht, B.; Yanagida, T.; Scholz, Tomáš; Ito, A.; Jiménez, J. A.; Brabec, Jan

    2010-01-01

    Roč. 48, č. 9 (2010), s. 3111-3116. ISSN 0095-1137 R&D Projects: GA MŠk LC522 Institutional research plan: CEZ:AV0Z60220518 Keywords : MOLECULAR EVIDENCE * PACIFIC SALMON * NIHONKAIENSE * Diphyllobothrium * multiplex PCR * the cytochrome c oxidase subunit 1 * mitochondrial DNA Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 4.220, year: 2010

  16. A PCR assay for gender assignment in dugong (Dugong dugon) and West Indian manatee (Trichechus manatus).

    Science.gov (United States)

    McHale, M; Broderick, D; Ovenden, J R; Lanyon, J M

    2008-05-01

    Gender assignment for some aquatic mammals in the field is difficult. Molecular sexing from tissue biopsies is possible as males are heterogametic. Here we describe a multiplex PCR assay that amplifies the male specific SRY gene and differentiates ZFX and ZFY gametologues in two sirenian species, dugong (Dugong dugon) and West Indian manatee (Trichechus manatus). The assay was validated with animals of known gender and proved accurate and robust to experimental failure. PMID:21585866

  17. PCR AS A DIAGNOSTIC TOOL FOR BRUCELLOSIS

    Science.gov (United States)

    Numerous PCR-based assays have been developed for the identification of Brucella to improve diagnostic capabilities. Collectively, the repertoire of assays addresses several aspects of the diagnostic process. For some purposes, the simple identification of Brucella is adequate (e.g. diagnosis of ...

  18. Handheld real-time PCR device.

    Science.gov (United States)

    Ahrberg, Christian D; Ilic, Bojan Robert; Manz, Andreas; Neužil, Pavel

    2016-01-26

    Here we report one of the smallest real-time polymerase chain reaction (PCR) systems to date with an approximate size of 100 mm × 60 mm × 33 mm. The system is an autonomous unit requiring an external 12 V power supply. Four simultaneous reactions are performed in the form of virtual reaction chambers (VRCs) where a ≈200 nL sample is covered with mineral oil and placed on a glass cover slip. Fast, 40 cycle amplification of an amplicon from the H7N9 gene was used to demonstrate the PCR performance. The standard curve slope was -3.02 ± 0.16 cycles at threshold per decade (mean ± standard deviation) corresponding to an amplification efficiency of 0.91 ± 0.05 per cycle (mean ± standard deviation). The PCR device was capable of detecting a single deoxyribonucleic acid (DNA) copy. These results further suggest that our handheld PCR device may have broad, technologically-relevant applications extending to rapid detection of infectious diseases in small clinics. PMID:26753557

  19. 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…

  20. Detection of Mycobacterium Tuberculosis by using PCR

    International Nuclear Information System (INIS)

    Polymerase Chain Reaction (PCR) procedure using three primary set derived from repetitive DNA sequence specific to mycobacteria was used to diagnose pathogenic Mycobacterium tuberculosis. The assay was specific for M. tuberculosis and could be used to detect the amount DNA less than 10-9g

  1. 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

  2. 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.

  3. A comparison of the effects of PCR inhibition in quantitative PCR and forensic STR analysis.

    Science.gov (United States)

    Funes-Huacca, Maribel E; Opel, Kerry; Thompson, Robyn; McCord, Bruce R

    2011-04-01

    In this paper we compare the effects of three representative PCR inhibitors using quantitative PCR (qPCR) and multiplex STR amplification in order to determine the effect of inhibitor concentration on allele dropout and to develop better ways to interpret forensic DNA data. We have used humic acid, collagen and calcium phosphate at different concentrations to evaluate the profiles of alleles inhibited in these amplifications. These data were correlated with previously obtained results from quantitative PCR including melt curve effects, efficiency changes and cycle threshold (Ct) values. Overall, the data show that there are two competing processes that result from PCR inhibition. The first process is a general loss of larger alleles. This appears to occur with all inhibitors. The second process is more sequence specific and occurs when the inhibitor binds DNA, altering the cycle threshold and the melt curve. This sequence-specific inhibition results in patterns of allele loss that occur in addition to the overall loss of larger alleles. The data demonstrate the applicability of utilizing real-time PCR results to predict the presence of certain types of PCR inhibition in STR analysis. PMID:21462225

  4. 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 commonly used fluorescent chemistries are SYBR® Green dyes and TaqMan®, Molecular Beacon or Scorpion probes. SYBR® Green is very simple to use and cost efficient. As SYBR® Green dye binds to any double-stranded DNA product, its success depends greatly on proper primer design. Many types of online primer design software are available, which can be used free of charge to design desirable SYBR® Green-based qPCR primers. This laboratory exercise is intended for those who have a fundamental background in PCR. It addresses the basic fluorescent chemistries of real-time PCR, the basic rules and pitfalls of primer design, and provides a step-by-step protocol for designing SYBR® Green-based primers with free, online software. PMID:21445907

  5. Droplet digital PCR, the new tool in HIV reservoir quantification?

    OpenAIRE

    De Spiegelaere, Ward; Kiselinova, Maja; Malatinkova, Eva; Pasternak, Alexander; Berkhout, Ben; Vandekerckhove, Linos

    2013-01-01

    Background: Digital PCR is a relatively old concept for absolute quantification of DNA using PCR, but recent technological developments allowed its wide use. The current state of art technique for performing digital PCR is based on microdroplet technology. Direct absolute quantification relieves the necessity of standard curves and increases assay accuracy. In addition, the end point PCR set-up allows higher assay flexibility and decreases quantitative bias due to variations in PCR efficiency...

  6. Protein Binding between PcrG-PcrV and PcrH-PopB/PopD Encoded by the pcrGVH-popBD Operon of the Pseudomonas aeruginosa Type III Secretion System

    OpenAIRE

    Allmond, Leonard R.; Karaca, Timur J.; Nguyen, Vinh N.; Nguyen, Thong; Wiener-Kronish, Jeanine P.; Sawa, Teiji

    2003-01-01

    Of the proteins encoded by the pcrGVH-popBD operon of the Pseudomonas aeruginosa type III secretion system, PcrG bound to PcrV and PcrH bound to PopB/PopD. In addition, Yersinia LcrG bound to PcrV, and Yersinia LcrH bound to PopD. The results imply a highly functional conservation of type III secretion between P. aeruginosa and Yersinia species.

  7. Enhancing the efficiency of a PCR using gold nanoparticles

    OpenAIRE

    Li, Min; Lin, Yu-Cheng; Wu, Chao-Chin; Liu, Hsiao-Sheng

    2005-01-01

    We found that the PCR could be dramatically enhanced by Au nanoparticles. With the addition of 0.7 nM of 13 nm Au nanoparticles into the PCR reagent, the PCR efficiency was increased. Especially when maintaining the same or higher amplification yields, the reaction time could be shortened, and the heating/cooling rates could be increased. The excellent heat transfer property of the nanoparticles should be the major factor in improving the PCR efficiency. Different PCR systems, DNA polymerases...

  8. Environmentally friendly animal litter

    Energy Technology Data Exchange (ETDEWEB)

    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.

  9. Precision animal breeding

    OpenAIRE

    Flint, A.P.F.; WOOLLIAMS, J. A.

    2007-01-01

    We accept that we are responsible for the quality of life of animals in our care. We accept that the activities of man affect all the living things with which we share this planet. But we are slow to realize that as a result we have a duty of care for all living things. That duty extends to the breeding of animals for which we are responsible. When animals are bred by man for a purpose, the aim should be to meet certain goals: to improve the precision with which breeding outcomes can be predi...

  10. Kinect driven facial animation

    OpenAIRE

    Ojeda Noda, Guillermo

    2016-01-01

    Kinect es un dispositivo que se presenta en el ámbito de la industria de la animación como una alternativa económica. Haciendo uso de él, este proyecto desarrolla una aplicación de animación facial que aplique las expresiones faciales del usuario a un modelo 3D. Nowadays, facial animation is a core part of the character animation industry. From movies to video games, facial animation is done by most companies with the help of expensive equipment that capture real people's facial expression...

  11. 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...

  12. Standing for Animals

    OpenAIRE

    Sunstein, Cass Robert

    1999-01-01

    From the legal point of view, there is nothing at all new or unfamiliar in the idea of "animal rights;" on the contrary, it is entirely clear that animals have legal rights. Indeed, the rise of legal rights for animals has been one of the most distinctive features of the last thirty years of federal statutory law. An investigation of the question of standing helps show that the real issues involve problems of enforcement and scope. Human beings often do and should have standing to protect ani...

  13. Comparison of Droplet Digital PCR and Quantitative PCR Assays for Quantitative Detection of Xanthomonas citri Subsp. citri.

    Science.gov (United States)

    Zhao, Yun; Xia, Qingyan; Yin, Youping; Wang, Zhongkang

    2016-01-01

    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; Pvalue of ddPCR technology in the diagnosis of plant disease and quarantine applications. PMID:27427975

  14. Comparison of Simultaneous Splenic Sample PCR with Blood Sample PCR for Diagnosis and Treatment of Experimental Ehrlichia canis Infection

    OpenAIRE

    Harrus, Shimon; Kenny, Martin; Miara, Limor; Aizenberg, Itzhak; Waner, Trevor; Shaw, Susan

    2004-01-01

    This report presents evidence that dogs recover from acute canine monocytic ehrlichiosis (CME) after 16 days of doxycycline treatment (10 mg/kg of body weight every 24 h). Blood PCR was as valuable as splenic aspirate PCR for early diagnosis of acute CME. Splenic aspirate PCR was, however, superior to blood PCR for the evaluation of ehrlichial elimination.

  15. Molecular analysis of dolphin morbillivirus: A new sensitive detection method based on nested RT-PCR.

    Science.gov (United States)

    Centelleghe, Cinzia; Beffagna, Giorgia; Zanetti, Rossella; Zappulli, Valentina; Di Guardo, Giovanni; Mazzariol, Sandro

    2016-09-01

    Cetacean Morbillivirus (CeMV) has been identified as the most pathogenic virus for cetaceans. Over the past three decades, this RNA virus has caused several outbreaks of lethal disease in odontocetes and mysticetes worldwide. Isolation and identification of CeMV RNA is very challenging in whales because of the poor preservation status frequently shown by tissues from stranded animals. Nested reverse transcription polymerase chain reaction (nested RT-PCR) is used instead of conventional RT-PCR when it is necessary to increase the sensitivity and the specificity of the reaction. This study describes a new nested RT-PCR technique useful to amplify small amounts of the cDNA copy of Cetacean morbillivirus (CeMV) when it is present in scant quantity in whales' biological specimens. This technique was used to analyze different tissues (lung, brain, spleen and other lymphoid tissues) from one under human care seal and seven cetaceans stranded along the Italian coastline between October 2011 and September 2015. A well-characterized, 200 base pair (bp) fragment of the dolphin Morbillivirus (DMV) haemagglutinin (H) gene, obtained by nested RT-PCR, was sequenced and used to confirm DMV positivity in all the eight marine mammals under study. In conclusion, this nested RT-PCR protocol can represent a sensitive detection method to identify CeMV-positive, poorly preserved tissue samples. Furthermore, this is also a rather inexpensive molecular technique, relatively easy to apply. PMID:27220282

  16. The application of PCR in the detection of mycotoxigenic fungi in foods

    Directory of Open Access Journals (Sweden)

    Ursula Konietzny

    2003-12-01

    Full Text Available It is estimated that 25 to 50% of the crops harvested worldwide are contaminated with mycotoxins. Because of the toxic and carcinogenic potential of mycotoxins, there is an urgent need to develop detection methods that are rapid and highly specific. The highly advanced physico-chemical methods for the analysis of mycotoxins in use, have the disadvantage that highly sophisticated clean-up and/or derivatization procedures must be applied. An alternative could be the detection of the mycotoxigenic moulds themselves, especially as molecular techniques have been introduced recently as powerful tools for detecting and identifying fungi. PCR methods for the detection of aflatoxigenic Aspergilli, patulin-producing Penicillum and trichothecene- as well as fumonisin-producing Fusaria strains have been described. The usefulness of the PCR methods developed so far to monitor quality and safety in the food an feed industry was already demonstrated. Thus, PCR may be applied to the screening of agricultural commodities for the absence of mycotoxin producers prior to or even after processing. Negative results in this assay indicate that a sample should be virtually free of mycotoxins. Only the positive samples left must be analyzed for the presence of mycotoxins using physico-chemical standard methods. This review does not only summarize the so far developed qualitative and quantitative PCR assays for the detection of mycotoxigenic fungi in agricultural commodities, foods and animal feeds, but describes also strategies to develop new specific PCR assays for such a detection.

  17. Measurement of lentiviral vector titre and copy number by cross-species duplex quantitative PCR.

    Science.gov (United States)

    Christodoulou, I; Patsali, P; Stephanou, C; Antoniou, M; Kleanthous, M; Lederer, C W

    2016-01-01

    Lentiviruses are the vectors of choice for many preclinical studies and clinical applications of gene therapy. Accurate measurement of biological vector titre before treatment is a prerequisite for vector dosing, and the calculation of vector integration sites per cell after treatment is as critical to the characterisation of modified cell products as it is to long-term follow-up and the assessment of risk and therapeutic efficiency in patients. These analyses are typically based on quantitative real-time PCR (qPCR), but as yet compromise accuracy and comparability between laboratories and experimental systems, the former by using separate simplex reactions for the detection of endogene and lentiviral sequences and the latter by designing different PCR assays for analyses in human cells and animal disease models. In this study, we validate in human and murine cells a qPCR system for the single-tube assessment of lentiviral vector copy numbers that is suitable for analyses in at least 33 different mammalian species, including human and other primates, mouse, pig, cat and domestic ruminants. The established assay combines the accuracy of single-tube quantitation by duplex qPCR with the convenience of one-off assay optimisation for cross-species analyses and with the direct comparability of lentiviral transduction efficiencies in different species. PMID:26202078

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

    OpenAIRE

    Nevar García Bastidas; Pablo Gutiérrez Sánchez; Mauricio Marín Montoya

    2013-01-01

    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 pru...

  19. INMUNODIAGNOSTICO DE FASCIOLOSIS BOVINA MEDIANTE ELISA Y WESTERN BLOT

    Directory of Open Access Journals (Sweden)

    Texia Gorman

    1998-01-01

    Full Text Available Se evaluó y caracterizó la respuesta inmune humoral de bovinos naturalmente infectados con Fasciola hepatica frente a un extracto total de antígenos de excreción-secreción (E-S y a una fracción antigénica semipurificada cromatográficamente (Immunodiagnosis of bovine fasciolosis was evaluated in terms of its sensitivity and specificity by ELISA and Western blot using a crude excretory-secretory preparation (C E-S and an E-S partially purified chromatographic fraction of Fasciola hepatica (<30 kDa as antigens. Serum samples from 52 bovines with natural fasciolosis, 18 without the infection and 48 bovines with hydatid cysts, all checked by post mortem examination, were tested as well. The sensitivity and specificity obtained by ELISA using the C E-S antigens were 53% and 100%, respectively. Likewise, when using the partially purified fraction, the sensitivity of ELISA test increased to 90% with a 100% specificity. The C E-S antigens reacted specifically with sera from infected bovines exhibiting bands of 14, 22, 27-39 and 37-38 kDa in Western blots. When the partially purified fraction was tested, the 28-30 kDa band was consistently detected by sera from all infected bovines and was absent in sera from uninfected as well as in sera from those with hydatid infection. These results indicate that F. hepatica presents different antigens, among which the 28-30 kDa fraction represents polypeptides with diagnostic potential that can be further purified and applied in the immunodiagnosis of bovine fasciolosis corroborating previous results recorded in other animal species

  20. Bottom dwelling animals: Benthos

    Digital Repository Service at National Institute of Oceanography (India)

    Ingole, B.S.

    . At the bottom/sediment dwelling animal communities are collectively termed as 'BENTHOS'. This extremely valuable component of the marine environment consumes the sediment organic matter from the overlying water column and effectively converts into benthic...

  1. 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.

  2. Animation of Antimicrobial Resistance

    Medline Plus

    Full Text Available ... Food and Drug Administration's (FDA's) Center for Veterinary Medicine (CVM) produced a nine-minute animation explaining how ... efforts are underway in both veterinary and human medicine to preserve the effectiveness of these drugs. One ...

  3. Retrospectives: Animal Spirits

    OpenAIRE

    Roger Koppl

    1991-01-01

    John Maynard Keynes argued that when the conditions for rational action are not present, people are driven by "animal spirits." This article briefly considers Keynes' argument, and the history of the term.

  4. [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. PMID:20669543

  5. Animation of Antimicrobial Resistance

    Medline Plus

    Full Text Available ... About FDA Contact FDA Browse by Product Area Product Areas back Food Drugs Medical Devices Radiation-Emitting Products Vaccines, Blood & Biologics Animal & Veterinary Cosmetics Tobacco Products

  6. Commonality among Fluoroquinolone-Resistant Sequence Type ST131 Extraintestinal Escherichia coli Isolates from Humans and Companion Animals in Australia▿†

    OpenAIRE

    Platell, Joanne L.; Cobbold, Rowland N.; Johnson, James R.; Heisig, Anke; Heisig, Peter; Clabots, Connie; Kuskowski, Michael A.; Trott, Darren J

    2011-01-01

    Escherichia coli sequence type 131 (ST131), an emergent multidrug-resistant extraintestinal pathogen, has spread epidemically among humans and was recently isolated from companion animals. To assess for human-companion animal commonality among ST131 isolates, 214 fluoroquinolone-resistant extraintestinal E. coli isolates (205 from humans, 9 from companion animals) from diagnostic laboratories in Australia, provisionally identified as ST131 by PCR, selectively underwent PCR-based O typing and ...

  7. Animal Models of Fibromyalgia

    OpenAIRE

    Nagakura, Yukinori; Ito, Hiroyuki; Shimizu, Yasuaki

    2012-01-01

    Animal models of disease states are valuable tools for developing new treatments and investigating underlying mechanisms. They should mimic the symptoms and pathology of the disease and importantly be predictive of effective treatments. Fibromyalgia is characterized by chronic widespread pain with associated co-morbid symptoms that include fatigue, depression, anxiety and sleep dysfunction. In this review, we present different animal models that mimic the signs and symptoms of fibromyalgia. T...

  8. Animal models of schizophrenia

    OpenAIRE

    Jones, CA; Watson, DJG; Fone, KCF

    2011-01-01

    Developing reliable, predictive animal models for complex psychiatric disorders, such as schizophrenia, is essential to increase our understanding of the neurobiological basis of the disorder and for the development of novel drugs with improved therapeutic efficacy. All available animal models of schizophrenia fit into four different induction categories: developmental, drug-induced, lesion or genetic manipulation, and the best characterized examples of each type are reviewed herein. Most rod...

  9. Laboratory animal allergy.

    OpenAIRE

    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 at 8 laboratory animal facilities. All participants completed a questionnaire and underwent skin prick testing with common and occupational allergens. Total and specific IgE measures were obtained....

  10. Whole animal imaging

    OpenAIRE

    Sandhu, Gurpreet Singh; Solorio, Luis; Broome, Ann-Marie; Salem, Nicolas; Kolthammer, Jeff; Shah, Tejas; Flask, Chris; Duerk, Jeffrey L.

    2010-01-01

    Translational research plays a vital role in understanding the underlying pathophysiology of human diseases, and hence development of new diagnostic and therapeutic options for their management. After creating an animal disease model, pathophysiologic changes and effects of a therapeutic intervention on them are often evaluated on the animals using immunohistologic or imaging techniques. In contrast to the immunohistologic techniques, the imaging techniques are noninvasive and hence can be us...

  11. Cultural - Amusement Animation

    OpenAIRE

    Jakovlev, Zlatko; Koteski, Cane; Angelkova, Tanja; Dzambazoski, Kristijan

    2011-01-01

    In the contemporary tourism, the cultural – amusing animation is at the same level as the food and the accommodation. This animation contributes to avoid the monotony and boredom of the guests/visitors.The visitors need diversion and dynamics during their tourist stay.There is nothing more destructive for themas the feeling of boredom, itself. The whole phenomenom of amusement and diversion is very relevant and characteristic for the human and it has its roots in the need for change, because ...

  12. 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.

  13. 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.

  14. 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....

  15. Experimental Animal Welfare

    OpenAIRE

    Yusuf Ergun

    2011-01-01

    It is an obvious obligation for investigators to consume millions of experimental animals every year to obtain scientific data. Because most of these experiments involve painful and distressing procedures, to obey the so-called 3Rs, reduction, refinement and replacement, is a prerequisite for those who would apply to ethics committees for a given research proposal. Of the 3Rs, refinement could be defined as “decrease in the incidence of severity of inhumane procedures applied to those animals...

  16. Small Animal Bone Biomechanics

    OpenAIRE

    Vashishth, Deepak

    2008-01-01

    Animal models, in particular mice, offer the possibility of naturally achieving or genetically engineering a skeletal phenotype associated with disease and conducting destructive fracture tests on bone to determine the resulting change in bone’s mechanical properties. Several recent developments, including nano- and micro- indentation testing, microtensile and microcompressive testing, and bending tests on notched whole bone specimens, offer the possibility to mechanically probe small animal ...

  17. Detection of hepatitis C virus RNA: comparison of one-stage polymerase chain reaction (PCR) with nested-set PCR.

    OpenAIRE

    Gretch, D R; Wilson, J. J; Carithers, R L; dela Rosa, C; Han, J. H.; Corey, L

    1993-01-01

    We evaluated a new hepatitis C virus RNA assay based on one-stage PCR followed by liquid hybridization with an oligonucleotide probe and compared it with nested-set PCR. The one-stage and nested-set PCR assays had identical sensitivities in analytical experiments and showed 100% concordance when clinical specimens were used. One-stage PCR may be less prone to contamination than nested-set PCR.

  18. Information Fusion with Belief Functions: a comparison of Proportional Conflict Redistribution PCR5 and PCR6 rules for Networked Sensors

    OpenAIRE

    Roman Ilin; Erik Blasch

    2015-01-01

    We compare several belief fusion methods, including the proportional conflict redistribution rules (PCR5 and PCR6) for multiple sources. The PCR fusion of evidence methods have shown improvement over the classical Dempster-Shafer and Bayesian fusion techniques in the presence of conflicting information. The PCR6 rule shows improvement over PCR5 when the number of sources increases. Using Hasse graphical diagrams, we highlight the comparison between the methods...

  19. 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

    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...... 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...

  20. 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.

  1. 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

    2016-08-01

    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. PMID:27503920

  2. Simulación de medios de transmisión mediante el método de elementos finitos

    OpenAIRE

    Revillas Sánchez, Carlos

    2015-01-01

    El análisis de medios de transmisión y componentes de circuitos de radiofrecuencia, como filtros, antenas, acopladores, etc. se puede realizar mediante instrumentación especifica, analizadores de red, osciloscopios, analizadores de espectro, o mediante métodos de computación electromagnética, realizando simulaciones para analizar los medios de transmisión y componentes de radiofrecuencia. El análisis mediante simulaciones presenta la clara ventaja de que no es necesario constru...

  3. Comparison of Droplet Digital PCR and Quantitative PCR Assays for Quantitative Detection of Xanthomonas citri Subsp. citri

    Science.gov (United States)

    Yin, Youping; Wang, Zhongkang

    2016-01-01

    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. PMID:27427975

  4. Animal, animalité, devenir-animal

    OpenAIRE

    Viennet, Denis

    2011-01-01

    Question de regard Nous regardons les animaux et les animaux nous regardent. Nous faisons signe à un chat, par la voix, par le geste, le chat nous regarde et cligne des yeux. Il n’a pas la capacité d’exprimer des paroles selon le modèle humain, mais à sa manière il nous répond, par un clin d’œil. Que se passe-t-il dans ce clin d’œil ? Une communication s’établit, un échange a lieu. Nous regardons l’animal qui nous regarde. Que voyons-nous alors ? Le clin d’œil énigmatique nous pousse à regard...

  5. 9 CFR 79.4 - Designation of scrapie-positive animals, high-risk animals, exposed animals, suspect animals...

    Science.gov (United States)

    2010-01-01

    ... complete a post exposure monitoring and management plan. Testing may include live-animal testing using a... testing of genetically susceptible animals in the flock that cannot be evaluated by a live animal test... investigation which may include additional testing of the suspect animal and or animals that have...

  6. PCR AS DIAGNOSTIC METHOD IN AQUACULTURE

    OpenAIRE

    Ivančica Strunjak-Perović; Natalija Topić Popović

    1999-01-01

    PCR is an acronym for »polymerase chain reaction«, a technique based on detection and amplification of specific DNA and RNA sequences. It can be applied in diagnostics of hereditary diseases, forensics, population genetics, systematics, bioengineering, evolution biology, and also aquaculture. With this method it is possible to diagnose an array of viral, bacterial and parasitic diseases of fish, shellfish and crustaceans. The advantages of the technique are manifested in rapid obtaining of re...

  7. PCR identification of Mycobacterium bovis BCG.

    OpenAIRE

    Talbot, E. A.; Williams, D L; Frothingham, R

    1997-01-01

    The attenuated bacillus Calmette-Guérin (BCG) vaccine strain is derived from a virulent strain of Mycobacterium bovis. BCG is difficult to differentiate from other strains of M. bovis and other members of the M. tuberculosis complex by conventional methods. Recently, a genomic region designated RD1 was found to be present in all virulent M. bovis and M. tuberculosis strains tested but deleted from all BCG strains tested. With this information, a multiplex PCR method was developed to detect th...

  8. QUANTATITIVE PCR ASSAY FOR MARINE BACTERIA

    OpenAIRE

    Brunk, Clifford F.

    2003-01-01

    Monitoring the bacterial flora in coastal marine waters by conventional techniques has been difficult as most of the bacteria do not readily grow on culture plates and their morphologies are virtually identical in the microscope. Molecular techniques, particularly characterizing bacteria using polymerase chain reaction (PCR) amplification of their small subunit ribosomal RNA (SSU rRNA) genes, has dramatically improved the ability to identify bacteria from environmental samples. Identificatio...

  9. Algorithms and Software for PCR Primer Design

    OpenAIRE

    Huang, Yu-ting

    2015-01-01

    Polymerase Chain Reaction (PCR) is a widely used technology in molecular bi-ology for DNA amplification. To generate multiple copies of a DNA molecule, a pair ofprimers (two synthesized DNA sequences with a total length of 15-30 bases) are annealedto the boundaries of the targeted DNA molecule. Then, the new replicated DNA fragmentelongates from one primer to the other.Though primers always hybridize to their respective complements within DNAsequences, primer pairs for targeted DNA sequences ...

  10. 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.

  11. 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)

  12. Comparison of noninvasive sample collection procedures for canine leishmaniasis diagnosis by PCR-hybridization

    International Nuclear Information System (INIS)

    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 32P 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 μL 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)

  13. Animal models of ADHD.

    Science.gov (United States)

    Bari, A; Robbins, T W

    2011-01-01

    Studies employing animal models of attention-deficit/hyperactivity disorder (ADHD) present clear inherent advantages over human studies. Animal models are invaluable tools for the study of underlying neurochemical, neuropathological and genetic alterations that cause ADHD, because they allow relatively fast, rigorous hypothesis testing and invasive manipulations as well as selective breeding. Moreover, especially for ADHD, animal models with good predictive validity would allow the assessment of potential new therapeutics. In this chapter, we describe and comment on the most frequently used animal models of ADHD that have been created by genetic, neurochemical and physical alterations in rodents. We then discuss that an emerging and promising direction of the field is the analysis of individual behavioural differences among a normal population of animals. Subjects presenting extreme characteristics related to ADHD can be studied, thereby avoiding some of the problems that are found in other models, such as functional recovery and unnecessary assumptions about aetiology. This approach is justified by the theoretical need to consider human ADHD as the extreme part of a spectrum of characteristics that are distributed normally in the general population, as opposed to the predominant view of ADHD as a separate pathological category. PMID:21287324

  14. Seguimiento continuo del proceso de gelificacion de alcoxidos de silicio y titanio mediante ensayos reologicos

    OpenAIRE

    Rudé i Payró, Elisabet; Llorens Llacuna, Joan; Mans Teixidó, Claudi, 1948-

    2000-01-01

    En este trabajo se ha estudiado el proceso de gelificación desde un nuevo punto de vista: el seguimiento de la transición solgel mediante la evolución con el tiempo de la distribución de tiempos de relajación, que está directamente relacionada con la distribución de pesos moleculares. La técnica utilizada para la determinación de la distribución de tiempos de relajación es la reología. El proceso de gelificación se ha seguido de forma continua mediante dos ensayos reológicos distintos: (1) en...

  15. 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. PMID:25889355

  16. Redes inalámbricas y simulación de WLAN mediante OPNET

    OpenAIRE

    Romero Kanashiro, Walter R.

    2013-01-01

    Proyecto que incluye una investigación sobre las tecnologías usadas en redes inalámbricas y su simulación mediante OPNET para su evaluación. Projecte que inclou una investigació sobre les tecnologies usades en xarxes sense fils i la seva simulació mitjançant OPNET per a la seva avaluació.

  17. Effetto dell'analgesia epidurale sulla progressione della testa fetale valutata mediante ecografia 3D

    OpenAIRE

    Arcangeli, Tiziana

    2014-01-01

    Introduzione: L'analgesia epidurale è stata messa in correlazione con l'aumento della durata del secondo stadio del travaglio e del tasso di utilizzo della ventosa ostetrica. Diversi meccanismi sono stati ipotizzati, tra cui la riduzione di percezione della discesa fetale, della forza di spinta e dei riflessi che promuovono la progressione e rotazione della testa fetale nel canale del parto. Tali parametri sono solitamente valutati mediante esame clinico digitale, costantemente riportato ...

  18. Terrestrial animal ecology

    International Nuclear Information System (INIS)

    The Animal Ecology project is an integral part of the terrestrial ecology program. For convenience, it is reported separately because of the specialized nature of its techniques. It includes studies to characterize faunal populations taxonomically and ecologically and to estimate density and biomass of important mammal, bird, herpetofauna, and invertebrate populations. Extensive studies of small mammal populations conducted in past years are being summarized for open literature publication. Methodology and techniques developed in the animal ecology program are expected to be vital to studies to be initiated under a newly funded 189 entitled Radioecology of Waste Management Zones. These kinds of supportive studies will be needed to determine dietary habits of important animals inhabiting waste management zones, construction of realistic food chain models, and estimating radioactivity doses to biota

  19. Statistics of lattice animals

    Science.gov (United States)

    Hsu, Hsiao-Ping; Nadler, Walder; Grassberger, Peter

    2005-07-01

    The scaling behavior of randomly branched polymers in a good solvent is studied in two to nine dimensions, modeled by lattice animals on simple hypercubic lattices. For the simulations, we use a biased sequential sampling algorithm with re-sampling, similar to the pruned-enriched Rosenbluth method (PERM) used extensively for linear polymers. We obtain high statistics of animals with up to several thousand sites in all dimension 2⩽d⩽9. The partition sum (number of different animals) and gyration radii are estimated. In all dimensions we verify the Parisi-Sourlas prediction, and we verify all exactly known critical exponents in dimensions 2, 3, 4, and ⩾8. In addition, we present the hitherto most precise estimates for growth constants in d⩾3. For clusters with one site attached to an attractive surface, we verify the superuniversality of the cross-over exponent at the adsorption transition predicted by Janssen and Lyssy.

  20. 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.

  1. 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)

  2. The experiments on healthy animals

    International Nuclear Information System (INIS)

    In this chapter author describes the experiments on leukotitin influence on hematosis which was held on :1. healthy animals received the preparation; 2. irradiated animals received the preparation; 3. irradiated animals didn't receive the preparation

  3. Companion animal adoption study.

    Science.gov (United States)

    Neidhart, Laura; Boyd, Renee

    2002-01-01

    To better understand the outcomes of companion animal adoptions, Bardsley & Neidhart Inc. conducted a series of 3 surveys over a 1-year period with dog and cat owners who had adopted their pet through either a (a) Luv-A-Pet location, (b) Adopt-a-thon, or (c) traditional shelter. This article suggests opportunities to improve owners' perceptions of their pets and the adoption process through (a) providing more information before adoption about pet health and behaviors, (b) providing counseling to potential adopters to place pets appropriately, and (c) educating adopters to promote companion animal health and retention. Results demonstrate that the pet's relationship to the family unit, such as where the pet sleeps and how much time is spent with the pet, is related to the amount of veterinary care the companion animal receives, and to long-term retention. Satisfaction and retention are attributed to the pet's personality, compatibility, and behavior, rather than demographic differences among adopters or between adoption settings. The age of the companion animal at adoption, the intended recipient, and presence of children in the home also play a role. Health problems were an issue initially for half of all adopted pets, but most were resolved within 12 months. Roughly one fourth of adopters who no longer have their companion animal said their pet died. Characteristics of pets that died support the contention that spaying and neutering profoundly affects a companion animal's life span. Although retention is similar for dogs and cats, mortality is higher among cats in the first year after adoption. PMID:12578739

  4. 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

  5. COMPAIXÃO ANIMAL

    OpenAIRE

    Márcio Seligmann Silva

    2011-01-01

    O trabalho estuda a questão da compaixão, que na história do pensamento foi ora tratada como uma marca da humanidade, ora pensada como uma marca de nossa origem natural e animal. Para Lactâncio, por exemplo, sem piedade o homem é um animal. O texto parte de uma discussão de Buffon, que falava de uma compaixão como uma de nossas “affections naturelles”. Para ele, “a alma tem menos a ver do que o corpo nesse sentimento de piedade natural e os animais, assim como o homem, sã...

  6. L’animal

    OpenAIRE

    Rongier, Sébastien

    2012-01-01

    On sait que l’amitié entre Maurice Blanchot et Emmanuel Lévinas est dense, complète et exigeante. À partir de quelques textes (et des lignes de fuite), je voudrais souligner ce lien amical et intellectuel, la constance avec laquelle les deux hommes ont mutuellement nourri leur réflexion, leur écriture. Et, au fil des lectures, s’est progressivement dégagée l’idée de l’animal comme espace d’interrogation, l’animal et l’animalité comme enjeu pour lire le lien, mais aussi la distance. Trois text...

  7. Animal models of tinnitus.

    Science.gov (United States)

    Brozoski, Thomas J; Bauer, Carol A

    2016-08-01

    Presented is a thematic review of animal tinnitus models from a functional perspective. Chronic tinnitus is a persistent subjective sound sensation, emergent typically after hearing loss. Although the sensation is experientially simple, it appears to have central a nervous system substrate of unexpected complexity that includes areas outside of those classically defined as auditory. Over the past 27 years animal models have significantly contributed to understanding tinnitus' complex neurophysiology. In that time, a diversity of models have been developed, each with its own strengths and limitations. None has clearly become a standard. Animal models trace their origin to the 1988 experiments of Jastreboff and colleagues. All subsequent models derive some of their features from those experiments. Common features include behavior-dependent psychophysical determination, acoustic conditions that contrast objective sound and silence, and inclusion of at least one normal-hearing control group. In the present review, animal models have been categorized as either interrogative or reflexive. Interrogative models use emitted behavior under voluntary control to indicate hearing. An example would be pressing a lever to obtain food in the presence of a particular sound. In this type of model animals are interrogated about their auditory sensations, analogous to asking a patient, "What do you hear?" These models require at least some training and motivation management, and reflect the perception of tinnitus. Reflexive models, in contrast, employ acoustic modulation of an auditory reflex, such as the acoustic startle response. An unexpected loud sound will elicit a reflexive motor response from many species, including humans. Although involuntary, acoustic startle can be modified by a lower-level preceding event, including a silent sound gap. Sound-gap modulation of acoustic startle appears to discriminate tinnitus in animals as well as humans, and requires no training or

  8. God, animals and zombies

    OpenAIRE

    Lynch, Joseph J.

    2011-01-01

    Argumentos neo-cartesianos recientes intentan reducir los animales a zombis filosóficos, seres sin estados de conciencia fenoménica. Si tales argumentos fuesen correctos, los animales verdaderamente no sufrirían, y, por tanto, no existiría el problema de Dios y el sufrimiento animal. En mi opinión, la afirmación de que los animales son zombis no es suficientemente plausible para proporcionar una teodicea adecuada acerca del problema de Dios y el dolor animal.

  9. 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.

  10. Laboratorio de sanidad animal

    OpenAIRE

    anonymous

    2009-01-01

    El laboratorio de Sanidad Animal de Jove, integrado orgánicamente en el CIATA, es el Laboratorio Oficial de Análisis en el campo de la Sanidad Animal del Principado de Asturias. Entre sus objetivos está el análisis y diagnóstico de las enfermedades animales con mayor interés sanitario y el apoyo técnico y análisis de muestras para la erradicación de la Brucelosis, Perineumonia Bovina y Leucosis Enzoótica bovina.

  11. 锚定PCR(Anchored PCR):一种新的染色体步行方法

    Institute of Scientific and Technical Information of China (English)

    陈柏君; 孙超; 王勇; 胡鸢雷; 林忠平

    2004-01-01

    基于PCR的技术是克隆已知DNA片段侧翼序列的最常用方法.到目前为止,这些方法大致可以分为3种类型:反向PCR(inverse PCR)、连接介导的PCR(1igation-mediated PCR)和随机引物PCR(randomly primed PCR).反向PCR是使用最早的方法,其原理是用限制性内切酶消化基因组总

  12. 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.

  13. Comparison of Nonculture Blood-Based Tests for Diagnosing Invasive Aspergillosis in an Animal Model

    OpenAIRE

    White, P. Lewis; Wiederhold, Nathan P.; Loeffler, Juergen; Najvar, Laura K.; Melchers, Willem; Herrera, Monica; Bretagne, Stephane; Wickes, Brian; Kirkpatrick, William R.; Barnes, Rosemary A.; Donnelly, J. Peter; Patterson, Thomas F.

    2016-01-01

    The European Aspergillus PCR Initiative (EAPCRI) has provided recommendations for the PCR testing of whole blood (WB) and serum/plasma. It is important to test these recommended protocols on nonsimulated “in vivo” specimens before full clinical evaluation. The testing of an animal model of invasive aspergillosis (IA) overcomes the low incidence of disease and provides experimental design and control that is not possible in the clinical setting. Inadequate performance of the recommended protoc...

  14. Nucleic acid extraction, oligonucleotide probes and PCR methods

    International Nuclear Information System (INIS)

    Complex microbiomes of rumen and gastrointestinal tracts. Bacteria, fungi and protozoa, present in rumen and gastrointestinal (GI) tracts, interact with feed, with each other, and with their host animals, resulting in a complex symbiotic microbiota of distinctive composition and structure. Such microbiota is dynamic and highly responsive to a variety of biotic and abiotic factors, such as diet, feed additives, age, health and physiological status of the host animal, geographical locations, season and feeding regimen (reviewed in Ref. [39]). This symbiotic microbiota has been the focus of microbial research for over half a century in search for improved ruminant nutrition. Before the advent of molecular biology techniques, microorganisms in rumen and GI tracts, as in other habitats, were studied with cultivation-based techniques, which only allows for the isolation and characterization of a limited number of readily culturable species. As estimated, there are more than 400 species of bacteria and up to 100 species of protozoa and fungi inhabiting rumen and GI tracts. In human GI tracts, as much as 60% of these members cannot be isolated on agar plates and, thus, remain unknown. In ruminants, although it is not known, the culturable species of the microbiota are probably in the same range. Even among the culturable species, probably only some of them have been isolated and described. The application of cultivation-independent, more sensitive and accurate molecular techniques to the study of ruminal and GI microorganisms provided an alternative to directly examining the diversity and the community structure of ruminal and GI microbiota on the basis of genotypes, instead of phenotypes. Both polymerase chain reaction (PCR)-based methods, such as denaturing gradient gel electrophoresis (DGGE), ribosomal intergenic spacer analysis, terminal restriction fragment length polymorphism, cloning and sequencing of PCR amplicons and amplified 16S ribosomal DNA restriction

  15. Imaging Histone Methylations in Living Animals.

    Science.gov (United States)

    Sekar, Thillai V; Paulmurugan, Ramasamy

    2016-01-01

    Histone modifications (methylation, acetylation, phosphorylation, sumoylation, etc.,) are at the heart of cellular regulatory mechanisms, which control expression of genes in an orderly fashion and control the entire cellular regulatory networks. Histone lysine methylation has been identified as one of the several posttranslational histone modifications that plays crucial role in regulating gene expressions in facultative heterochromatic DNA regions while maintaining structural integrity in constitutive heterochromatic DNA regions. Since histone methylation is dysregulated in various cellular diseases, it has been considered a potential therapeutic target for drug development. Currently there is no simple method available to screen and preclinically evaluate drugs modulating this cellular process, we recently developed two different methods by adopting reporter gene technology to screen drugs and to preclinically evaluate them in living animals. Method detects and quantitatively monitors the level of histone methylations in intact cells, is of a prerequisite to screen small molecules that modulate histone lysine methylation. Here, we describe two independent optical imaging sensors developed to image histone methylations in cells and in living animals. Since we used standard PCR-based cloning strategies to construct different plasmid vectors shown in this chapter, we are not providing any details regarding the construction methods, instead, we focus on detailing various methods used for measuring histone methylation-assisted luciferase quantitation in cells and imaging in living animals. PMID:27424907

  16. Presence of Clostridium difficile PCR ribotype clusters related to 033, 078 and 045 in diarrhoeic calves in Germany.

    Science.gov (United States)

    Schneeberg, Alexander; Neubauer, Heinrich; Schmoock, Gernot; Grossmann, Ernst; Seyboldt, Christian

    2013-08-01

    This study provides data on the distribution and relationship of C. difficile PCR ribotypes in diarrhoeic calves in Germany. C. difficile was isolated from 176 of 999 (17.6 %) faecal samples or swabs of diarrhoeic calves from 603 farms collected between January 2010 and August 2012 by eight federal laboratories of six states. Strains were assigned to 17 PCR ribotypes. PCR ribotypes 033 (57 %), 078 (17 %) and 045/FLI01 (closest match to 045 in the WEBRIBO database; 9 %) were found the most frequently. Nine per cent of all culture-positive tested animals shed more than one multiple locus variable number tandem repeat analysis (MLVA) or PCR ribotype. Eight PCR ribotypes with related profiles (including 033, 078 and 045/FLI01) representing 92 % of all isolates were grouped into three clusters. Molecular relatedness was supported by the absence of the MLVA locus A6Cd only in clustered strains and identical toxin gene profiles for strains within each cluster. Previously reported mulitilocus sequence typing analysis for PCR ribotypes that were also recovered in this study found identical sequence types and a tcdC deletion (Δ39 bp) for 033, 045, 078 and 126 (ST-11), confirming this clustering. A different geographical occurrence of PCR ribotypes was shown for cluster 033 (found more frequently in southern Germany) and 045 (found more frequently in northern Germany). This study showed that clusters of C. difficile PCR ribotypes related to 033, 078 and 045 are predominant in diarrhoeic calves in Germany. The high number of strains belonging to PCR ribotype 078 demonstrated that diarrhoeic calves are also potential reservoirs for human pathogenic C. difficile strains. PMID:23639987

  17. Monitorización de una central fotovoltaica mediante sensores inalámbricos con tecnología Bluetooth

    OpenAIRE

    Abajo González, David

    2015-01-01

    Este proyecto consiste en utilizar tecnología inalámbrica para monitorizar un parque fotovoltaico. Se trata de monitorizar una central fotovoltaica mediante sensores inalámbricos con tecnología zigbee. Se ha realizado una posible planificación para el parque la cual se ha basado únicamente en datos obtenidos mediante simulación, sería una buena idea poder comparar estos datos en un futuro con datos de mediciones reales.

  18. Geolocalizaciones newtoniana y postnewtoniana mediante la técnica TDOA: el efecto "time-delay" de Shapiro

    OpenAIRE

    Rodríguez Teijeiro, María del Carmen

    2011-01-01

    En esta Tesis se presenta un procedimiento para localizar radiotransmisores pasivos situados en el entorno de la Tierra mediante satélites artificiales y mediciones TDOA. Las localizaciones de los radiotransmisores se determinan en forma cerrada mediante ecuaciones algebraicas que corresponden a los modelos newtoniano y postnewtoniano del campo gravitatorio terrestre. La Geolocalización, es decir, la localización de un radiotransmisor cuyas señales son captadas por un conjunto de receptores e...

  19. Sensitive ligand-based protein quantification using immuno-PCR

    DEFF Research Database (Denmark)

    Hansen, Marcus Celik; Nederby, Line; Henriksen, Mads Okkels-Birk; Hansen, Maria; Nyvold, Charlotte Guldborg

    2014-01-01

    Quantitative PCR (qPCR) of reverse-transcribed mRNA has revolutionized gene expression analyses. qPCR analysis is based on the prevalent assumption that mRNA transcript numbers provide an adequate measure of specific biomarker expression. However, taking the complexity of protein turnover into...... account, there is a need to correlate qPCR-derived transcriptional patterns with protein translational patterns so as to not leave behind important pathobiological details. One emerging approach in protein analysis is PCR-coupled protein quantification, often denoted as immuno-PCR (iPCR), which targets...... soluble proteins. Here we review recent trends and applications in iPCR assays that may bridge the gap between classical enzyme-linked immunosorbent assays and mass spectrometry methodologies in terms of sensitivity and multiplexing....

  20. Multiplex polymerase chain reaction (PCR) on a SU-8 chip

    DEFF Research Database (Denmark)

    Christensen, Troels Balmer; Bang, Dang Duong; Wolff, Anders

    2008-01-01

    We present the detection of Campylobacter at species level using multiplex PCR in a micro fabricated PCR chip. The chip is based on the polymer SU-8 that allows integration with different microfluidic components, e.g., sample pre-treatment before PCR, and DNA detection simultaneously with or after...... the PCR. The chip performs very well with respect to heating and cooling rates with values up to around 40 °C/s and 20 °C/s, respectively, and has low power consumption (0.5–2.5 W depending on temperature). Multiplex DNA amplification by PCR for the detection of Campylobacter at species level...... was performed successfully on the chip with results comparable to conventional PCR methods. Microsystems often show serious PCR inhibition due to a high surface to volume ratio which causes an increased proclivity of the PCR mix ingredients to stick to the surfaces. To avoid this, a simple method of dynamic...

  1. DNA fingerprinting of medically important microorganisms by use of PCR.

    OpenAIRE

    van Belkum, A

    1994-01-01

    Selected segments of any DNA molecule can be amplified exponentially by PCR. This technique provides a powerful tool to detect and identify minimal numbers of microorganisms. PCR is applicable both in diagnosis and in epidemiology. By amplification of hypervariable DNA domains, differences can be detected even among closely related strains. PCR fingerprinting is a valuable tool for medical microbiologists, epidemiologists, and microbial taxonomists. The current state of PCR-mediated genotypin...

  2. In vivo cloning of PCR products in E. coli.

    OpenAIRE

    Oliner, J D; Kinzler, K W; Vogelstein, B

    1993-01-01

    This report describes an efficient method to clone PCR products exploiting endogenous Escherichia coli enzymatic activities. PCR products are engineered to contain terminal sequences identical to sequences at the two ends of a linearized vector. PCR products and vector DNA are then simply co-transfected into E. coli strain JC8679, obviating the requirement for enzymatic treatment of the PCR product or in vitro ligation. The high rate of homologous recombination in this strain results in effic...

  3. PCR-based diagnosis of human fungal infections

    OpenAIRE

    Khot, Prasanna D.; Fredricks, David N

    2009-01-01

    PCR is a very appealing technology for the detection of human pathogens, but the detection of fungal pathogens is particularly challenging. Fungi have cell walls that impede the efficient lysis of organisms and liberation of DNA, which can lead to false-negative PCR results. Conversely, some human pathogens are also ubiquitous environmental saprophytes that can contaminate PCR reagents and cause false-positive results. We examine the quality of PCR-based studies for fungal diagnostics using 4...

  4. Clinical Utility of Droplet Digital PCR for Human Cytomegalovirus

    OpenAIRE

    Sedlak, Ruth Hall; Cook, Linda; Cheng, Anqi; Magaret, Amalia; Jerome, Keith R.

    2014-01-01

    Human cytomegalovirus (CMV) has historically been the major infectious cause of morbidity and mortality among patients receiving hematopoietic cell or organ transplant. Standard care in a transplant setting involves frequent monitoring of CMV viral load over weeks to months to determine when antiviral treatment may be required. Quantitative PCR (qPCR) is the standard molecular diagnostic method for monitoring. Recently, digital PCR (dPCR) has shown promise in viral diagnostics, although curre...

  5. 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.

  6. Freeing Captive Animals

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Even though theymight not haveenough food intheir own stom-achs,Tibetan peasantswould feed their draughtcattle with the best food,asthey depended on them forplowing. Such good treat-ment lasted until the ani-mals died,after which,some peasants would burythem in their own fields,

  7. 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...

  8. 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...

  9. 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...

  10. 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

  11. Counting Hexagonal Lattice Animals

    OpenAIRE

    Mohammed, Mohamud

    2002-01-01

    We describe Maple packages for the automatic generation of generating functions(and series expansions) for counting lattice animals(fixed polyominoes), in the two-dimensional hexagonal lattice, of bounded but arbitrary width. Our Maple packages(complete with source code) are easy-to-use and available from my website.

  12. Holographic Animation Apparatus.

    Science.gov (United States)

    Johnston, Sean F.

    1979-01-01

    Describes a simple apparatus for producing strip holograms with a number of slit-shaped exposures displaced along the vertical direction. The hologram maintains full horizontal parallax, but the slit aperture reduces the vertical viewing angle of the animated object. (Author/GA)

  13. 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.

  14. Animation-based Sketching

    DEFF Research Database (Denmark)

    Vistisen, Peter

    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 be...

  15. Animation of Antimicrobial Resistance

    Medline Plus

    Full Text Available ... back to top Popular Content Home Latest Recalls Report an Adverse Event MedWatch Safety Alerts News Releases Consumer Updates About FDA Contact FDA Browse by Product Area Product Areas back Food Drugs Medical Devices Radiation-Emitting Products Vaccines, Blood & Biologics Animal & ...

  16. Animal ethics dilemma

    DEFF Research Database (Denmark)

    Dich, Trine; Hansen, Tina; Algers, Anne;

    2006-01-01

    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 to...

  17. Impactor No More (Animation)

    Science.gov (United States)

    2005-01-01

    [figure removed for brevity, see original site] Quick Time Movie for PIA02130 Realtime Ejecta (Animation) This movie was taken by Deep Impact's flyby spacecraft shows the flash that occurred when comet Tempel 1 ran over the spacecraft's probe. It was taken by the flyby craft's medium-resolution camera.

  18. From Spirit's Perspective (Animation)

    Science.gov (United States)

    2004-01-01

    This animation shows the perspective from the navigation camera on the Mars Exploration Rover Spirit before and after its automated stand-up process. After standing up, the rover is approximately 12 inches higher off of the lander, resulting in a better view of the surrounding terrain.

  19. 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.

  20. 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

  1. 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.

  2. Comparison of a Commercially Available Repetitive-Element PCR System (DiversiLab) with PCR Ribotyping for Typing of Clostridium difficile Strains ▿

    OpenAIRE

    Eckert, C; Van Broeck, J; Spigaglia, P.; Burghoffer, B; Delmée, M; Mastrantonio, P; Barbut, F.

    2011-01-01

    This study compared a repetitive-element PCR (rep-PCR) method (DiversiLab system) to PCR ribotyping. The discriminatory power of rep-PCR was 0.997. Among the PCR ribotype 027 isolates tested, different rep types could be distinguished. rep-PCR showed a higher discriminatory power than PCR ribotyping. Nevertheless, this method requires technical skill, and visual interpretation of rep-PCR fingerprint patterns may be difficult.

  3. 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…

  4. People vs. animals.

    Science.gov (United States)

    Engram, S

    1992-07-12

    Animal rights activists demonstrated against physicians in Pittsburgh, Pennsylvania, who had transplanted a baboon liver into a man. They complained that baboons should not serve as spare parts for humans, but the complaint misfired when another man with liver disease challenged them. Nevertheless the rapidly growing population in the world is threatening animal species such as elephants. In Zimbabwe where a severe drought exists and which has been somewhat able to protect animals from poachers, the government now allows people to kill elephants and other animals for their meat. The great numbers of wildlife have placed considerable population pressure on Gonarezhou National Park. The government hopes the good will plan will reduce the number of illegal poachings in the future. This illustrates the need for population stability to protect the environment. Yet the 1992 UN environment conference in Rio de Janeiro, Brazil, did not address population growth as a threat to biodiversity and the environment. Indeed if population continues to grow at its present rate, the population in 2100 will stand at 19 billion and each year before that the Earth will lose more farmland and forests and witness more days of smog, polluted water, political instabilities, and environmental refugees. Viruses like HIV may afflict the population. Most of the population growth will be in developing countries where drought and economic and political instabilities are common. In 2100 with such a hugh population, a national park for wildlife will most likely only be a luxury. We can no longer be complacent and must take action now to prevent this disaster. It will soon be clear that a growing population does not produce more prosperity as many economists would like us to believe, and discussions about using animals for spare parts will be ludicrous. PMID:12286283

  5. Multiplex PCR serogrouping of Listeria monocytogenes isolated in Japan.

    Science.gov (United States)

    Shimojima, Yukako; Ida, Miki; Nishino, Yukari; Ishitsuka, Rie; Kuroda, Sumiyo; Hirai, Akihiko; Sadamasu, Kenji; Nakama, Akiko; Kai, Akemi

    2016-04-01

    PCR serogrouping methods were used to examine strains of L. monocytogenes isolated in Japan. Among 187 strains, 99.5% were classified into 4 PCR serogroups corresponding to conventional serotypes. Only one isolate had a new PCR profile, which may be a variant of serogroup IVb. PMID:26537550

  6. Real-Time PCR for Universal Phytoplasma Detection and Quantification

    DEFF Research Database (Denmark)

    Christensen, Nynne Meyn; Nyskjold, Henriette; Nicolaisen, Mogens

    2013-01-01

    Currently, the most efficient detection and precise quantification of phytoplasmas is by real-time PCR. Compared to nested PCR, this method is less sensitive to contamination and is less work intensive. Therefore, a universal real-time PCR method will be valuable in screening programs and in other...

  7. Determining Fungi rRNA Copy Number by PCR

    Science.gov (United States)

    The goal of this project is to improve the quantification of indoor fungal pollutants via the specific application of quantitative PCR (qPCR). Improvement will be made in the controls used in current qPCR applications. This work focuses on the use of two separate controls within ...

  8. Development and Evaluation of a Real-Time PCR Assay for Detection and Quantification of Blastocystis Parasites in Human Stool Samples: Prospective Study of Patients with Hematological Malignancies▿

    OpenAIRE

    Poirier, Philippe; Wawrzyniak, Ivan; Albert, Aurélie; El Alaoui, Hicham; Delbac, Frédéric; Livrelli, Valérie

    2011-01-01

    Blastocystis anaerobic parasites are widespread worldwide in the digestive tract of many animal species, including humans. Epidemiological Blastocystis studies are often limited by the poor sensitivity of standard parasitological assays for its detection. This report presents a highly sensitive real-time quantitative PCR (qPCR) assay developed to detect Blastocystis parasites in stool samples. The assay targets a partial sequence of the Blastocystis small ribosomal subunit (SSU) rRNA gene, al...

  9. PCR AS DIAGNOSTIC METHOD IN AQUACULTURE

    Directory of Open Access Journals (Sweden)

    Ivančica Strunjak-Perović

    1999-12-01

    Full Text Available PCR is an acronym for »polymerase chain reaction«, a technique based on detection and amplification of specific DNA and RNA sequences. It can be applied in diagnostics of hereditary diseases, forensics, population genetics, systematics, bioengineering, evolution biology, and also aquaculture. With this method it is possible to diagnose an array of viral, bacterial and parasitic diseases of fish, shellfish and crustaceans. The advantages of the technique are manifested in rapid obtaining of results, high specificity and sensitivity.

  10. Analysis of extracellular RNA by digital PCR

    Directory of Open Access Journals (Sweden)

    Kenji eTakahashi

    2014-06-01

    Full Text Available The transfer of extracellular RNA is emerging as an important mechanism for intracellular communication. The ability for the transfer of functionally active RNA molecules from one cell to another within vesicles such as exosomes enables a cell to modulate cellular signaling and biological processes within recipient cells. The study of extracellular RNA requires sensitive methods for the detection of these molecules. In this methods article, we will describe protocols for the detection of such extracellular RNA using sensitive detection technologies such as digital PCR. These protocols should be valuable to researchers interested in the role and contribution of extracellular RNA to tumor cell biology.

  11. Attitudes towards animal use and belief in animal mind

    OpenAIRE

    Knight, Sarah; Vrij, Aldert; Cherryman, Julie; Nunkoosing, Karl

    2004-01-01

    Animals are used by humans in many ways, yet science has paid little attention to the study of human-animal relationships (Melson 2002). In the present study participants (n= 96) completed a questionnaire on attitudes towards animal use and individual differences were examined to determine which characteristics might underlie these attitudes (‘belief in animal mind’, age, gender, experience of animals, vegetarianism, political stance, and living area). It emerged that participants held differ...

  12. Design of Multiplex Polymerase Chain Reaction (PCR Method for Molecular Detection of Yersinia pestis Bacterium

    Directory of Open Access Journals (Sweden)

    Mohammad Soleimani

    2010-01-01

    Full Text Available Objective: Yersinia pestis, the causative agent of the zoonotic plague infection, is a majorpublic health concern both as a threat and potential bioweapon. The objective of thepresent study was to establish a uniplex and multiplex - polymerase chain reaction (PCRtest for the specific detection of Y. pestis.Materials and Methods: PCR reactions performed by three pair primers which targetedthe caf1 and pla genes located on the pFra and pPst plasmids and the irp2 chromosomalgene located on the ‘pathogenicity island’. After TA cloning of the PCR products, the test’slimit of detection (LOD was determined. For evaluating the specificity, PCR reactionswere performed with negative control bacteria.Results: Assays were performed with the genome of Y. pestis which produced three DNAfragments of the expected sizes 300, 400 and 520 bp which corresponded to the irp2,caf1 and pla genes, respectively. The lower LoD was 370 copy numbers for the caf1 geneand 21 for the pla gene. In PCR reactions that used negative control bacteria, detectablefragments were not observed.Conclusion: Our method clearly discriminated Y. pestis DNA. The rapidity, specificityand sensitivity of this procedure suggest that it can serve as a useful alternative methodfor the inoculation of laboratory animals or the use of specific culture media for routineplaque surveillance and outbreak investigations. Another vital result of this study was theestablishment of Y. pestis molecular detection technique in Iran.

  13. An efficient multistrategy DNA decontamination procedure of PCR reagents for hypersensitive PCR applications.

    Directory of Open Access Journals (Sweden)

    Sophie Champlot

    Full Text Available BACKGROUND: PCR amplification of minute quantities of degraded DNA for ancient DNA research, forensic analyses, wildlife studies and ultrasensitive diagnostics is often hampered by contamination problems. The extent of these problems is inversely related to DNA concentration and target fragment size and concern (i sample contamination, (ii laboratory surface contamination, (iii carry-over contamination, and (iv contamination of reagents. METHODOLOGY/PRINCIPAL FINDINGS: Here we performed a quantitative evaluation of current decontamination methods for these last three sources of contamination, and developed a new procedure to eliminate contaminating DNA contained in PCR reagents. We observed that most current decontamination methods are either not efficient enough to degrade short contaminating DNA molecules, rendered inefficient by the reagents themselves, or interfere with the PCR when used at doses high enough to eliminate these molecules. We also show that efficient reagent decontamination can be achieved by using a combination of treatments adapted to different reagent categories. Our procedure involves γ- and UV-irradiation and treatment with a mutant recombinant heat-labile double-strand specific DNase from the Antarctic shrimp Pandalus borealis. Optimal performance of these treatments is achieved in narrow experimental conditions that have been precisely analyzed and defined herein. CONCLUSIONS/SIGNIFICANCE: There is not a single decontamination method valid for all possible contamination sources occurring in PCR reagents and in the molecular biology laboratory and most common decontamination methods are not efficient enough to decontaminate short DNA fragments of low concentration. We developed a versatile multistrategy decontamination procedure for PCR reagents. We demonstrate that this procedure allows efficient reagent decontamination while preserving the efficiency of PCR amplification of minute quantities of DNA.

  14. 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.

  15. Diagnostic evaluation of a multiplexed RT-PCR microsphere array assay for the detection of foot-and-mouth disease virus and look-alike disease viruses

    Energy Technology Data Exchange (ETDEWEB)

    Hindson, B J; Reid, S M; Baker, B R; Ebert, K; Ferris, N P; Bentley Tammero, L F; Lenhoff, R J; Naraghi-Arani, P; Vitalis, E A; Slezak, T R; Hullinger, P J; King, D P

    2007-07-26

    A high-throughput multiplexed assay was developed for the differential laboratory diagnosis of foot-and-mouth disease virus (FMDV) from viruses which cause clinically similar diseases of livestock. This assay simultaneously screens for five RNA and two DNA viruses using multiplexed reverse transcription PCR (mRT-PCR) amplification coupled with a microsphere hybridization array and flow-cytometric detection. Two of the seventeen primer-probe sets included in this multiplex assay were adopted from previously characterized real-time RT-PCR (rRT-PCR) assays for FMDV. The diagnostic accuracy of the mRT-PCR was evaluated using 287 field samples, including 248 (true positive n= 213, true negative n=34) from suspect cases of foot-and-mouth disease collected from 65 countries between 1965 and 2006 and 39 true negative samples collected from healthy animals. The mRT-PCR assay results were compared with two singleplex rRT-PCR assays, using virus isolation with antigen-ELISA as the reference method. The diagnostic sensitivity of the mRT-PCR assay for FMDV was 93.9% [95% C.I. 89.8-96.4%], compared to 98.1% [95% C.I. 95.3-99.3%] for the two singleplex rRT-PCR assays used in combination. In addition, the assay could reliably differentiate between FMDV and other vesicular viruses such as swine vesicular disease virus and vesicular exanthema of swine virus. Interestingly, the mRT-PCR detected parapoxvirus (n=2) and bovine viral diarrhea virus (n=2) in clinical samples, demonstrating the screening potential of this mRT-PCR assay to identify viruses in FMDV-negative material not previously recognized using focused single-target rRT-PCR assays.

  16. Diagnosis of canine visceral leishmaniasis with radiolabelled probes: comparison of the kDNA PCR-hybridization with three molecular methods in different clinical samples

    International Nuclear Information System (INIS)

    Leishmania (Leishmania) chagasi is responsible for visceral leishmaniasis (VL) in Brazil and the dog is the main domestic reservoir. Disease control is based on the elimination of infected animals and the use of a sensitive and specific diagnostic test is necessary. The Brazilian VL control program emphasizes serologic surveys, mainly using the enzyme-linked immunosorbent assay (ELISA) and the immunofluorescence antibody test (IFAT), followed by the elimination of the seropositive dogs. However, these techniques present limitations in terms of sensitivity and specificity. The Polymerase Chain Reaction (PCR) associated to hybridization with DNA probes labeled with 32P has been recognized as a valuable tool for Leishmania identification. In this study, the sensitivity of kDNA PCR hybridization method was compared with three other molecular methods: Internal Transcribed Spacer 1 Nested PCR (ITS-1nPCR), Leishmania nested PCR (LnPCR) and Seminested kDNA PCR (kDNA snPCR). The comparison was performed in different clinical specimens: conjunctival swab, skin, blood and bone marrow. A group of thirty symptomatic dogs, positive in the parasitological and serological tests, was used. When. The techniques targeting kDNA mini-circles (kDNA snPCR and KDNA PCR-hybridization) showed the worst result for blood samples. The KDNA-PCR hybridization showed the best sensitivity for conjunctival swab. By comparing the samples on the basis of positivity obtained by the sum of all methods, the blood showed the worst outcome (71/120).The bone marrow showed the highest positivity (106/120), followed by conjunctival swab (100/120) and skin (89/120). Since the bone marrow samples are unsuitable for routine epidemiological surveys, the conjunctival swab was recommended because it allows high sensitivity, especially when associated with kDNA PCR hybridization method, and is a noninvasive sampling method. (author)

  17. Diagnosis of canine visceral leishmaniasis with radiolabelled probes: comparison of the kDNA PCR-hybridization with three molecular methods in different clinical samples

    Energy Technology Data Exchange (ETDEWEB)

    Ferreira, Aline Leandra C.; Ferreira, Sidney A.; Carregal, Virginia M.; Andrade, Antero Silva R., E-mail: antero@cdtn.br [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil). Lab. de Radiobiologia; Melo, Maria N., E-mail: melo@icb.ufmg.br [Departamento de Parasitologia. Instituto de Ciencias Biologicas. Universidade Federal de Minas Gerais, Belo Horizonte, MG (Brazil)

    2011-07-01

    Leishmania (Leishmania) chagasi is responsible for visceral leishmaniasis (VL) in Brazil and the dog is the main domestic reservoir. Disease control is based on the elimination of infected animals and the use of a sensitive and specific diagnostic test is necessary. The Brazilian VL control program emphasizes serologic surveys, mainly using the enzyme-linked immunosorbent assay (ELISA) and the immunofluorescence antibody test (IFAT), followed by the elimination of the seropositive dogs. However, these techniques present limitations in terms of sensitivity and specificity. The Polymerase Chain Reaction (PCR) associated to hybridization with DNA probes labeled with {sup 32}P has been recognized as a valuable tool for Leishmania identification. In this study, the sensitivity of kDNA PCR hybridization method was compared with three other molecular methods: Internal Transcribed Spacer 1 Nested PCR (ITS-1nPCR), Leishmania nested PCR (LnPCR) and Seminested kDNA PCR (kDNA snPCR). The comparison was performed in different clinical specimens: conjunctival swab, skin, blood and bone marrow. A group of thirty symptomatic dogs, positive in the parasitological and serological tests, was used. When. The techniques targeting kDNA mini-circles (kDNA snPCR and KDNA PCR-hybridization) showed the worst result for blood samples. The KDNA-PCR hybridization showed the best sensitivity for conjunctival swab. By comparing the samples on the basis of positivity obtained by the sum of all methods, the blood showed the worst outcome (71/120).The bone marrow showed the highest positivity (106/120), followed by conjunctival swab (100/120) and skin (89/120). Since the bone marrow samples are unsuitable for routine epidemiological surveys, the conjunctival swab was recommended because it allows high sensitivity, especially when associated with kDNA PCR hybridization method, and is a noninvasive sampling method. (author)

  18. Development of a pan-Babesia FRET-qPCR and a survey of livestock from five Caribbean islands

    OpenAIRE

    Li, Jing; Kelly, Patrick; Zhang, Jilei; Xu, Chuanling; Wang, Chengming

    2015-01-01

    Background Babesia spp. are tick-borne protozoan hemoparasites and the second most common blood-borne parasites of mammals, in particular domestic animals. We used the Clustal Multiple Alignment program and 18S rRNA gene sequences of 22 Babesia species from GenBank to develop a PCR that could detect a wide variety of Babesia spp. in a single reaction. The pan-Babesia FRET-qPCR we developed reliably detected B. gibsoni, B. canis, B. vogeli, B. microti, B. bovis, and B. divergens under controll...

  19. 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, ...

  20. Amp-PCR: Combining a Random Unbiased Phi29-Amplification with a Specific Real-Time PCR, Performed in One Tube to Increase PCR Sensitivity

    OpenAIRE

    Lena Erlandsson; Lars Peter Nielsen; Anders Fomsgaard

    2010-01-01

    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 sam...

  1. An Assessment of Whole Blood and Fractions by Nested PCR as a DNA Source for Diagnosing Canine Ehrlichiosis and Anaplasmosis

    Directory of Open Access Journals (Sweden)

    Tereza Emmanuelle de Farias Rotondano

    2012-01-01

    Full Text Available Ehrlichiosis and anaplasmosis are tick-borne diseases. Ehrlichia canis and Anaplasma platys infect mainly white cells and platelets, respectively. The main DNA source for PCR is peripheral blood, but the potential of blood cell fractions has not been extensively investigated. This study aims at assessment of whole blood (WB and blood fractions potential in nested PCR (nPCR to diagnose canine ehrlichiosis and anaplasmosis. The 16S rRNA gene was amplified in 71.4, 17.8, 31.57, and 30% of the WB, granulocyte (G, mononuclear cells (M, and buffy coat (BC samples. Compared to the WB, the sensitivity of the PCR was 42.86% for the M, and BC fractions, 21.43% for the G, and 33.33% for the blood clot (C. There was fair agreement between the WB and M, BC and C, and slight with the G. Fair agreement occurred between the nPCR and morulae in the blood smear. One animal was coinfected with A. platys and E. canis. This study provided the first evidence of A. platys infection in dogs in Paraíba, Brazil, and demonstrated that WB is a better DNA source than blood fractions to detect Ehrlichia and Anaplasma by nPCR, probably because of the plasma bacterial concentration following host cell lysis.

  2. Comparative analysis of cultural isolation and PCR based assay for detection of Campylobacter jejuni in food and faecal samples

    Directory of Open Access Journals (Sweden)

    Harkanwaldeep Singh

    2011-03-01

    Full Text Available In the present study, the efficacy of polymerase chain reaction (PCR based on mapA gene of C. jejuni was tested for detection of Campylobacter jejuni in naturally infected as well as spiked faecal and food samples of human and animal origin. Simultaneously, all the samples were subjected to the cultural isolation of organism and biochemical characterization. The positive samples resulted in the amplification of a DNA fragment of size ~589 bp in PCR assay whereas the absence of such amplicon in DNA extracted from E. coli, Listeria, Salmonella and Staphylococcus confirmed the specificity of the primers. Of randomly collected 143 faecal samples comprising human diarrheic stools (43, cattle diarrheic faeces (48 and poultry faecal swabs (52 only 4, 3 and 8, respectively, could be detected by isolation whereas 6, 3 and 10, respectively, were found positive by PCR. However, among food samples viz. beef (30, milk (35, cheese (30, only one beef sample was detected both by culture as well as PCR. Additionally, PCR was found to be more sensitive for C. jejuni detection in spiked faecal and food samples (96.1% each as relative to culture isolation which could detect the organism in 86.7% and 80% samples, respectively. The results depicted the superior efficacy of PCR for rapid screening of samples owing to its high sensitivity, specificity and automation potential.

  3. Identification of periodontopathogen microorganisms by PCR technique

    Directory of Open Access Journals (Sweden)

    Milićević Radovan

    2008-01-01

    Full Text Available INTRODUCTION Periodontitis is an inflammatory disease of the supporting tissues of teeth and is a major cause of tooth loss in adults. The onset and progression of periodontal disease is attributed to the presence of elevated levels of a consortium of pathogenic bacteria. Gram negative bacteria, mainly strict anaerobes, play the major role. OBJECTIVE The present study aimed to assess the presence of the main types of microorganisms involved in the aetiopathogenesis of periodontal disease: Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Eikenella corrodens, Treponema denticola, Tanerella forsythia and Prevotella intermedia in different samples collected from the oral cavity of 90 patients diagnosed with periodontitis. METHOD Bacterial DNA detection was performed in diverse biological materials, namely in dental plaque, gingival tissue and saliva, by means of multiplex PCR, a technique that allows simultaneous identification of two different bacterial genomes. RESULTS In the dental plaque of the periodontitis patients, Treponema denticola dominated. In the gingival tissue, Tannerella forsythia and Treponema denticola were the microbiota most frequently detected, whilst in saliva Treponema denticola and Eikenella corrodens were found with the highest percentage. CONCLUSION The identification of microorganisms by multiplex PCR is specific and sensitive. Rapid and precise assessment of different types of periodontopathogens is extremely important for early detection of the infection and consequently for the prevention and treatment of periodontal disease. In everyday clinical practice, for routine bacterial evaluation in patients with periodontal disease, the dental plaque is the most suitable biological material, because it is the richest in periodontal bacteria.

  4. Cryptococcus species identification by multiplex PCR.

    Science.gov (United States)

    Leal, Ana Lusia; Faganello, Josiane; Bassanesi, Maria Cristina; Vainstein, Marilene H

    2008-06-01

    Members of the Cryptococcus species complex are encapsulated basidiomycetous yeasts, which can affect the central nervous system (CNS) and if untreated may cause meningitis. Cryptococcus neoformans is an opportunistic pathogen causing infections mainly in immunocompromised individuals. Cryptococcus gattii is a primary pathogen responsible for a high incidence of cryptococcomas in the lung and brain and shows a delayed response to antifungal therapy. The differentiation between the two species is primarily based on their growth on and color change of canavanine - glycine-bromothymol blue agar (CGB). Since this test is not always reliable, a multiplex PCR to identify both Cryptococcus species using more than 130 samples was standardized and the results obtained compared to those with the CGB test, using the Crypto Check serotyping kit as the standard. The multiplex PCR was shown to be more specific than the CGB test, in that results obtained with it were in agreement with those from serotyping all the samples, while the data from the CGB test disagreed with 6 out of 131 samples. PMID:18415847

  5. 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.

  6. Weak-willed Animals?

    Directory of Open Access Journals (Sweden)

    Thomas Spitzley

    2009-01-01

    Full Text Available The aim of this paper is to contribute to answering the conceptual question whether there can be weak-willed non-human animals. After some preliminary clarifications concerning the phenomenon of weakness of will three different accounts are examined for the conditions a being has to fulfill in order to be in a position to display weakness of will. It is argued that these conditions are very strong and that there are good reasons to assume that, e.g., only language users can be weak-willed. This is taken as an independent argument for Davidson's thesis that non-human animals which are not language users cannot act intentionally.

  7. 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...... by an empirical study of Responsible Research and Innovation (RRI) in a Triple Helix constellation. Using a three-week long innovation workshop, U- CrAc, involving 16 Danish companies and organisations and 142 students as empirical data, we discuss how animation-based sketching can explore not yet existing user...... 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...

  8. 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.

  9. 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.

  10. [Influenza in heterothermic animals].

    Science.gov (United States)

    Mancini, Dalva Assunção Portari; Mendonça, Rita Maria Zucatelli; Cianciarullo, Aurora Marques; Kobashi, Leonardo Setsuo; Trindade, Hermínio Gomes; Fernandes, Wilson; Pinto, José Ricardo

    2004-01-01

    The objective was to study Orthomyxovirus in heterothermic animals. Blood samples from snakes (genus Bothrops and Crotalus) and from toads and frogs (genus Bufo and Rana) were collected to evaluate the red cell receptors and antibodies specific to influenza virus by the hemagglutination and hemagglutination inhibition tests, respectively. Both snakes and toads kept in captivity presented receptors in their red cells and antibodies specific to either influenza virus type A (human and equine origin) or influenza type B. The same was observed with recently captured snakes. Concerning the influenza hemagglutination inhibition antibodies protective levels were observed in the reptiles' serum, against influenza type A and type B. Unlike the toads, 83.3% of the frogs presented mean levels of Ab 40HIU for some influenza strains. It was concluded that heterothermic animals could offer host conditions to the influenza virus and also susceptibility to the infection. PMID:15330057

  11. Modeling animal landscapes.

    Science.gov (United States)

    Porter, W P; Ostrowski, S; Williams, J B

    2010-01-01

    There is an increasing need to assess the effects of climate and land-use change on habitat quality, ideally from a mechanistic basis. The symposium "Molecules to Migration: Pressures of Life" at the Fourth International Conference in Africa for Comparative Physiology and Biochemistry, Maasai Mara National Reserve, Kenya, 2008, illustrated how the principles of biophysical ecology can capture the mechanistic links between organisms, climate, and other habitat features. These principles provide spatially explicit assessments of habitat quality from a physiological perspective (i.e., "animal landscapes") that can be validated independently of the data used to derive and parameterize them. The contents of this symposium showcased how the modeling of animal landscapes can be used to assess key issues in applied and theoretical ecology. The presentations included applications to amphibians, reptiles, birds, and mammals. The rare Arabian oryx on the Arabian Peninsula is used as an example for energetic calculations and their implications for behavior on the landscape. PMID:20670170

  12. 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

    OpenAIRE

    Jensen Jørgen; Østergaard Lars; Birkebæk Niels H; Birkelund Svend; Mygind Tina; Christiansen Gunna

    2002-01-01

    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. pneumon...

  13. IS711-based real-time PCR assay as a tool for detection of Brucella spp. in wild boars and comparison with bacterial isolation and serology

    Directory of Open Access Journals (Sweden)

    Miserez Raymond

    2009-07-01

    Full Text Available Abstract Background Control of brucellosis in livestock, wildlife and humans depends on the reliability of the methods used for detection and identification of bacteria. In the present study, we describe the evaluation of the recently established real-time PCR assay based on the Brucella-specific insertion sequence IS711 with blood samples from 199 wild boars (first group of animals and tissue samples from 53 wild boars (second group of animals collected in Switzerland. Results from IS711 real-time PCR were compared to those obtained by bacterial isolation, Rose Bengal Test (RBT, competitive ELISA (c-ELISA and indirect ELISA (i-ELISA. Results In the first group of animals, IS711 real-time PCR detected infection in 11.1% (16/144 of wild boars that were serologically negative. Serological tests showed different sensitivities [RBT 15.6%, c-ELISA 7.5% and i-ELISA 5.5%] and only 2% of blood samples were positive with all three tests, which makes interpretation of the serological results very difficult. Regarding the second group of animals, the IS711 real-time PCR detected infection in 26% of animals, while Brucella spp. could be isolated from tissues of only 9.4% of the animals. Conclusion The results presented here indicate that IS711 real-time PCR assay is a specific and sensitive tool for detection of Brucella spp. infections in wild boars. For this reason, we propose the employment of IS711 real-time PCR as a complementary tool in brucellosis screening programs and for confirmation of diagnosis in doubtful cases.

  14. Animal traction in Ghana:

    OpenAIRE

    Houssou, Nazaire; Kolavalli, Shashidhara; Bobobee, Emmanuel; Owusu, Victor

    2013-01-01

    The recent interest of the government of Ghana in agricultural mechanization has largely focused on the provision of tractors and imported machinery to the farming population. Animal traction has not received much attention from the country’s policymakers. The strong demand for mechanization services (Houssou et al., 2012; Benin et al., 2012) and inadequate number of tractors to meet the demand in the country call for more effective use of other power sources for the agriculture sector. Usi...

  15. Animal models of sepsis

    OpenAIRE

    Fink, Mitchell P.

    2013-01-01

    Sepsis remains a common, serious, and heterogeneous clinical entity that is difficult to define adequately. Despite its importance as a public health problem, efforts to develop and gain regulatory approval for a specific therapeutic agent for the adjuvant treatment of sepsis have been remarkably unsuccessful. One step in the critical pathway for the development of a new agent for adjuvant treatment of sepsis is evaluation in an appropriate animal model of the human condition. Unfortunately, ...

  16. Animal Models of Atherosclerosis

    OpenAIRE

    Godfrey S Getz; Reardon, Catherine A

    2012-01-01

    Atherosclerosis is a chronic inflammatory disorder that is the underlying cause of most cardiovascular disease. Both cells of the vessel wall and cells of the immune system participate in atherogenesis. This process is heavily influenced by plasma lipoproteins, genetics and the hemodynamics of the blood flow in the artery. A variety of small and large animal models have been used to study the atherogenic process. No model is ideal as each has its own advantages and limitations with respect to...

  17. 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.

  18. Metacognition in animals

    OpenAIRE

    Crystal, Jonathon D.; Foote, Allison L.

    2009-01-01

    Metacognition is thinking about thinking. There is considerable interest in developing animal models of metacognition to provide insight about the evolution of mind and a basis for investigating neurobiological mechanisms of cognitive impairments in people. Formal modeling of low-level (i.e., alternative) mechanisms has recently demonstrated that prevailing standards for documenting metacognition are inadequate. Indeed, low-level mechanisms are sufficient to explain data from existing methods...

  19. 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...

  20. AnimalChange

    OpenAIRE

    Van den Pol-van Dasselaar, Agnes; Bellocchi, Gianni; Hutchings, Nicholas John; Olesen, Jørgen Eivind; Saetnan, Eli Rudinow

    2014-01-01

    The EU-FP7 project AnimalChange (AN Integration of Mitigation and Adaptation options for sustainable Livestock production under climate CHANGE, http://www.animalchange.eu, 2011-2015) addresses mitigation and adaptation options and provides scientific guidance for their integration in sustainable development pathways for livestock production under climate change in Europe, Northern and Sub-Saharan Africa, and Latin America. The project provides insights, innovations, tools and models for lives...

  1. Slaughter - not only about animals

    OpenAIRE

    Wiberg, Sofia

    2012-01-01

    In order to get meat for human consumption animals have to be slaughtered. In Sweden, about 450,000 cattle are slaughtered every year; in 2011 93% of these were slaughtered at the 16 largest slaughter plants. Maintaining acceptable animal welfare standards in the industrial slaughter of animals places great demands on the management and staff. Good animal welfare means that consideration has been given to the animals' biology and subjective experience and to its possibilities to adapt to the ...

  2. Transfer to animals

    International Nuclear Information System (INIS)

    Data have been compiled to derive animal product transfer coefficients for radionuclides to update the values given in Technical Reports Series No. 364. Significant new data inputs have been incorporated from an extensive review of Russian language information and inclusion of data published since the early 1990s. The resultant database has been used to provide reference transfer coefficient values for a range of radionuclides to (i) cow, sheep and goat milk, (ii) meat (muscle) of cattle, sheep, goats, pigs and poultry and (iii) eggs. The approaches and procedures used to identify and collate data, and assumptions used are given. For most animal products, transfer coefficient values for elements additional to those in Technical Reports Series No. 364 are provided, although some elements were considered in the earlier evaluation which were not included in this review. Differences between the Technical Reports Series No. 364 'expected' values and the reference values from this document, which will be incorporated into the revised transfer parameter handbook, are discussed. An alternative approach to quantifying transfer by using concentration ratios is evaluated and CR values which could be applied across animal species have been provided for milk and meat. Information on fractional gastrointestinal absorption in adult ruminants has been compiled and reference values presented. Despite these improvements many data gaps remain. (author)

  3. 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.

  4. Novel polyomavirus detected in the feces of a chimpanzee by nested broad-spectrum PCR.

    Science.gov (United States)

    Johne, Reimar; Enderlein, Dirk; Nieper, Hermann; Müller, Hermann

    2005-03-01

    In order to screen for new polyomaviruses in samples derived from various animal species, degenerated PCR primer pairs were constructed. By using a nested PCR protocol, the sensitive detection of nine different polyomavirus genomes was demonstrated. The screening of field samples revealed the presence of a new polyomavirus, tentatively designated chimpanzee polyomavirus (ChPyV), in the feces of a juvenile chimpanzee (Pan troglodytes). Analysis of the region encoding the major capsid protein VP1 revealed a unique insertion in the EF loop of the protein and showed that ChPyV is a distinct virus related to the monkey polyomavirus B-lymphotropic polyomavirus and the human polyomavirus JC polyomavirus. PMID:15731285

  5. Comparative analysis of whole genome structure of Streptococcus suis using whole genome PCR scanning

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    An outbreak associated with Streptococcus suis infection in humans emerged in Sichuan province, China in 2005. The outbreak is atypical for the apparent large number of human cases, high fatality rate and geographical spread. To determine whether the bacterium has changed, we compared both human and animal isolates from the Sichuan outbreak with those collected previously within China and in other countries using whole genome PCR scanning (WGPScaning) comparative sequencing of several known virulence factor genes and multilocus sequence typing (MLST) analysis. WGPScanning analysis showed that all primer pairs yielded PCR products of the expected sizes in all four strains tested. The nucleotide sequences of all the detected virulence factor genes are identical in the four strains and MLST results showed that the four isolates studied and reference strain all belonged to the ST1 com-plex. No new genetic changes were found in the genome structure of the isolates from this Sichuan outbreak.

  6. Comparative analysis of whole genome structure of Streptococcus suis using whole genome PCR scanning

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    An outbreak associated with Streptococcus suis infection in humans emerged in Sichuan province, China in 2005. The outbreak is atypical for the apparent large number of human cases, high fatality rate and geographical spread. To determine whether the bacterium has changed, we compared both human and animal isolates from the Sichuan outbreak with those collected previously within China and in other countries using whole genome PCR scanning (WGPScaning) comparative sequencing of several known virulence factor genes and multilocus sequence typing (MLST) analysis. WGPScanning analysis showed that all primer pairs yielded PCR products of the expected sizes in all four strains tested. The nucleotide sequences of all the detected virulence factor genes are identical in the four strains and MLST results showed that the four isolates studied and reference strain all belonged to the ST1 complex. No new genetic changes were found in the genome structure of the isolates from this Sichuan outbreak.

  7. Application of qPCR assays for diagnosing causes of viral mink diarrhea. Preliminary results

    DEFF Research Database (Denmark)

    Hartby, Christina Marie; Kvisgaard, Lise Kirstine; Larsen, Lars Erik;

    ). Diarrhea in mink can be caused by infectious agents (virus, bacteria and parasites) and food-related/multifactorial conditions. Known enteric viral infections are mink enteritis virus (MEV) and mink astrovirus. Coronaviruses and caliciviruses have also been implicated as potential causes or contributors to...... diarrhea in mink. Rotavirus is poorly described in mink, but has previously been demonstrated in feces from mink pups with and without clinical signs (Jorgensen et al. 1996). The pathogenicity of these viruses could be related to viral load, virulence and the age of the mink. Therefore, there is a need for...... a quantitative diagnostic approach. We have developed new or adapted previously published real-time PCR/RT-PCR assays for MEV, astrovirus, rota- and coronavirus diagnostics. The technical test validation was initially carried out on archived diarrhea samples from diagnosed positive animals and on...

  8. 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. PMID:26845298

  9. A RAPID PCR-QUALITY DNA EXTRACTION METHOD IN FISH

    Institute of Scientific and Technical Information of China (English)

    LI Zhong; LIANG Hong-Wei; ZOU Gui-Wei

    2012-01-01

    PCR has been a general preferred method for biological research in fish, and previous research have enabled us to extract and purify PCR-quality DNA templates in laboratories[1-4]. The same problem among these procedures is waiting for tissue digesting for a long time. The overabundance time spent on PCR-quality DNA extraction restricts the efficiency of PCR assay, especially in large-scale PCR amplification, such as SSR-based genetic-mapping construction [5,6], identification of germ plasm resource[7,8] and evolution research [9,10], etc. In this study, a stable and rapid PCR-quality DNA extraction method was explored, using a modified alkaline lysis protocol. Extracting DNA for PCR only takes approximately 25 minutes. This stable and rapid DNA extraction method could save much laboratory time and promotes.%PCR has been a general preferred method for biological research in fish,and previous research have enabled us to extract and purify PCR-quality DNA templates in laboratories [1-4].The same problem among these procedures is waiting for tissue digesting for a long time.The overabundance time spent on PCR-quality DNA extraction restricts the efficiency of PCR assay,especially in large-scale PCR amplification,such as SSR-based genetic-mapping construction [5,6],identification of germ plasm resource[7,8] and evolution research [9,10],etc.In this study,a stable and rapid PCR-quality DNA extraction method was explored,using a modified alkaline lysis protocol.Extracting DNA for PCR only takes approximately 25 minutes.This stable and rapid DNA extraction method could save much laboratory time and promotes.

  10. 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%. PMID:25976998

  11. PCR and real-time PCR assays to detect fungi of Alternaria alternata species.

    Science.gov (United States)

    Kordalewska, Milena; Brillowska-Dąbrowska, Anna; Jagielski, Tomasz; Dworecka-Kaszak, Bożena

    2015-01-01

    Fungi of the Alternaria genus are mostly associated with allergic diseases. However, with a growing number of immunocompromised patients, these fungi, with A. alternata being the most prevalent one, are increasingly recognized as etiological agents of infections (phaeohyphomycoses) in humans. Nowadays, identification of Alternaria spp. requires their pure culture and is solely based on morphological criteria. Clinically, Alternaria infections may be indistinguishable from other fungal diseases. Therefore, a diagnostic result is often delayed or even not achieved at all. In this paper we present easy to perform and interpret PCR and real-time PCR assays enabling detection of A. alternata species. On the basis of alignment of β-tubulin gene sequences, A. alternata-specific primers were designed. DNA from fungal isolates, extracted in a two-step procedure, were used in PCR and real-time PCR assays followed by electrophoresis or melting temperature analysis, respectively. The assays specificity was confirmed, since positive results were obtained for all A. alternata isolates, and no positive results were obtained neither for other molds, dermatophytes, yeast-like fungi, nor human DNA. The assays developed here enable fast and unambiguous identification of A. alternata pathogens. PMID:26610309

  12. 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

  13. Knowledge of the Animal Welfare Act and Animal Welfare Regulations Influences Attitudes toward Animal Research

    OpenAIRE

    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...

  14. From the 'cinematic' to the 'anime-ic': Issues of movement in anime

    OpenAIRE

    Ruddell, C

    2008-01-01

    This is the author's accepted manuscript. The final published article is available from the link below. This article explores the way that movement is formally depicted in anime. Drawing on Thomas Lamarre's concepts of the `cinematic' and the `anime-ic', the article interrogates further the differences in movement and action in anime from traditional filmic form. While often considered in terms of `flatness', anime offers spectacle, character development and, ironically, depth through the ...

  15. Latent class analysis of the diagnostic characteristics of PCR and conventional bacteriological culture in diagnosing intramammary infections caused by Staphylococcus aureus in dairy cows at dry off

    DEFF Research Database (Denmark)

    Cederlöf, Sara Ellinor; Toft, Nils; Aalbæk, Bent;

    2012-01-01

    characteristics of PathoProof TM Mastitis PCR Assay and bacteriological culture (BC) in diagnosing bovine intramammary infections caused by S. aureus at dry off at different PCR cycle threshold (Ct)-value cut-offs. METHODS: Sterile quarter samples and non-sterile composite samples from 140 animals in seven herds...... were collected in connection with the dairy herd improvement (DHI) milk recording. All quarter samples were analyzed using BC whereas all composite samples were analyzed with PathoProof TM Mastitis PCR Assay. Latent class analysis was used to estimate test properties for PCR and BC in the absence of a...

  16. Detection of Human Papillomavirus DNA in Cervical Samples: Analysis of the New PGMY-PCR Compared To the Hybrid Capture II and MY-PCR Assays and a Two-Step Nested PCR Assay

    OpenAIRE

    Giovannelli, Lucia; Lama, Anna; Capra, Giuseppina; Giordano, Viviana; Aricò, Pietro; Ammatuna, Pietro

    2004-01-01

    The PGMY-PCR for human papillomavirus (HPV) was evaluated, in parallel with nested PCR (nPCR), in samples with noted Hybrid Capture II (HCII) and MY-PCR results. PGMY-PCR detected HPV DNA in 2.5% of HCII-negative-MY-PCR-negative samples and in 71.7% of HCII-positive-MY-PCR-negative samples; also, it detected the MY-PCR-negative-nPCR-negative types HPV-42, HPV-44, HPV-51, HPV-87, and HPV-89.

  17. PcrG protects the two long helical oligomerization domains of PcrV, by an interaction mediated by the intramolecular coiled-coil region of PcrG

    OpenAIRE

    Basu, Abhishek; Das, Urmisha; Dey, Supratim; Datta, Saumen

    2014-01-01

    Background PcrV is a hydrophilic translocator of type three secretion system (TTSS) and a structural component of the functional translocon. C-terminal helix of PcrV is essential for its oligomerization at the needle tip. Conformational changes within PcrV regulate the effector translocation. PcrG is a cytoplasmic regulator of TTSS and forms a high affinity complex with PcrV. C-terminal residues of PcrG control the effector secretion. Result Both PcrV and PcrG-PcrV complex exhibit elongated c...

  18. Experiencia en el Laboratorio de Matemática mediante el soporte de computadoras

    OpenAIRE

    Guala, G.; Oscherov, V.; Perez Millán, C.

    2009-01-01

    Nuestro propósito es presentar una experiencia desarrollada en el marco del Proyecto de Investigación: El uso de nuevas tecnologías en la enseñanza del cálculo. En él enfocamos el uso de nuevas tecnologías como un modo de superar la metodología habitual de dar las respuestas antes de que surjan las preguntas. Incorporamos el uso de un Laboratorio de Matemática mediante el soporte de computadoras para el que tuvimos en cuenta tanto las aplicaciones de la Matemática como la exigencia actual de ...

  19. Estudio mediante elementos finitos de los esfuerzos producidos en una biela de bicicleta

    OpenAIRE

    Gesé Bordils, Francisco Javier

    2013-01-01

    En el presente Trabajo Fin de Grado se realiza un modelo de un conjunto de pedalier de una bicicleta en un sistema CAD de manera que pueda importarse mediante un programa de elementos finitos y realizar simulaciones de cargas para obtener los estados de tensión y deformación de las piezas. Con la enorme evolución que se ha llevado a cabo en el mundo del ciclismo en las últimas décadas, especialmente a causa de la aparición de nuevos materiales y más aún en las técnicas de prototipado y ens...

  20. Monitorización de un lecho fluidizado mediante acelerometría

    OpenAIRE

    Velasco Fernández, Mario

    2013-01-01

    El presente proyecto estudiará la posibilidad de monitorizar un reactor químico mediante sensores de vibración. Actualmente, no se realiza este tipo de monitorización sobre reactores químicos, y los estudios realizados al respecto son escasos. Se tratarán de establecer las posibles equivalencias entre las medidas realizadas con sensores de presión y de vibración. Para ello se realizará la monitorización de un modelo de reactor a escala, del laboratorio de la Universidad, utilizand...