Yaima Burgher

Université de Montréal

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Evelyn Lobo

Centro Nacional de Sanidad Agropecuaria

Carl A. Gagnon

Université de Montréal

Arianna Duque Ortiz

Centro Nacional de Sanidad Agropecuaria

Chantale Provost

Agence Nationale de Sécurité Sanitaire de l’Alimentation, de l’Environnement et du Travail

Yoesley Lozada

Centro Nacional de Sanidad Agropecuaria

Marie-Jeanne Pesant

Université de Montréal

Jorge Timenetsky

Universidade de São Paulo

Anisleidy Pérez Castillo

Universidade Federal de Minas Gerais

Julia Noda

Centro Nacional de Sanidad Agropecuaria

Ana María Acevedo

University of Missouri

Cooperative Institutions

Centro Nacional de Sanidad Agropecuaria

Université de Montréal

Ministry of Agriculture, Fisheries and Food

Instituto de Medicina Tropical “Pedro Kourí”

Instituto de Investigaciones Avícolas

Universidade de São Paulo

Ministère de l'Agriculture

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Pan American Health Organization (Cuba)

Instituto Nacional de Higiene, Epidemiología y Microbiología

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DETECCIÓN EN MUESTRA CLÍNICA E IDENTIFICACIÓN DE AISLADOS DEL VIRUS DE LA BRONQUITIS INFECCIOSA AVIAR POR UN ENSAYO DE REVERSO TRANSCRIPCIÓN ACOPLADO A REACCIÓN EN CADENA DE LA POLIMERASA

Ana María Acevedo Yaima Burgher María Pilar Colás Bravo Damaris Relova A P Folmer Corrêa

La bronquitis infecciosa aviar (BIA) es una enfermedad viral ampliamente diseminada en el mundo y se considera una de las principales causas de perdidas economicas significativas en la industria avicola, debido a que afecta el desarrollo de aves de engorde y ponedoras. En Cuba, el diagnostico actual se realiza por aislamiento viral y es dificil su identificacion por metodos convencionales. Para identificar la posible presencia del virus de la BIA en aislados cubanos se aplico un ensayo de reverso transcripcion acoplado a reaccion en cadena de la polimerasa (RT-PCR) especifico para este virus. Las muestras de traquea-pulmon fueron tomadas como grupos de tres y cuatro aves despues de la necropsia de cada una, evaluandose un total de once aves. Las secuencias del virus de la BIA fueron detectadas en dos de los tres grupos evaluados. En uno de estos grupos se logro ademas, la deteccion del virus a partir de la muestra clinica; aspecto importante para la diferenciacion con enfermedades graves que cursan con un cuadro clinico similar como la Influenza aviar y la Enfermedad de Newcastle. La RT-PCR mostro potencial para ser usado en la identificacion de aislados del virus de la BIA, asi como en el examen directo a partir de muestras clinicas, lo que aporta posibilidades de su empleo en nuestro pais para la deteccion rapida, sensible y especifica de este agente, aspecto esencial en el diagnostico y control de esta infeccion viral.

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Virus Role During Intraepidemic Increase in Dengue Disease Severity

Vector-Borne and Zoonotic Diseases (2011)

R. Rodríguez Lizet Sánchez Yaima Burgher Delfina Rosario Mayling Álvarez

Dengue epidemics in Cuba have repeatedly demonstrated a month-to-month increase in clinical severity during secondary infections. The dengue 2 outbreak that occurred in Santiago de Cuba in 1997 was accompanied by the most severe intraepidemic increase in disease severity reported to date. It was initially proposed that the appearance of neutralization escape mutants during the course of the epidemic might explain this phenomenon. Recent studies have revealed that during the course of this epidemic, nucleotide substitutions appeared only in nonstructural (NS) genes, most of which were silent, except for one change in the NS1 gene. To study whether or not variation in the NS1 gene might be associated with increased disease severity during the epidemic, this gene was partially sequenced from 15 isolates obtained at different times during the 1997 epidemic. Early epidemic isolates differed from those obtained later by replacement only of threonine with serine at position 164 in the NS1 protein, an amino acid rarely found in any genotype of dengue 2 virus. All viruses isolated from patients located in Health Districts, where dengue 2 transmissions occurred late in the epidemic, contained Serine at position 164, indicating that this change was fixed within a few months. Here we argue that this single mutation contributes to viral survival or replication efficiency, resulting in enhanced infection in the presence of enhancing antibodies, a phenomenon that we term increased virus "fitness" in contrast to "virulence," an intrinsic property of the virus.

10.1089/vbz.2010.0177

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Detection in clinical sample and identification of avian infectious bronchitis virus isolates by a reverse transcription polymerasechain reaction assay.

Ana María Acevedo Yaima Burgher María Pilar Colás Bravo Damaris Relova Arisel Correa

Avian infectious bronchitis

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Desarrollo de PCR especie-específicos para la detección de micoplasmas contaminantes de cultivos celulares

Revista de salud animal (2020)

Anisleidy Pérez Castillo Arianna Duque Ortiz Yaima Burgher Jose Antonio Agüero Fernández Mario Sergio Solis Castells

Mollicute contamination in cell cultures is common. Species such as Mycoplasma arginini , Mycoplasma salivarium , Mycoplasma fermentans , Mycoplasma hyorhinis , Mycoplasma orale , and Acholeplasma laidlawii are identified as causing 95 % of the contaminations. The present work is aimed at developing species-specific PCRs for the identification of the most frequent contaminating mycoplasma species in cell cultures. The critical parameters of PCR were optimized and the analytical sensitivity and specificity were determined. The optimum temperature for hybridization was set at 55oC; magnesium concentration at 1,5 mM. A range of 0,3-0,5 µM was selected for the primers. The analytical sensitivity of PCR was 1 pg/µL, while its analytical specificity was 100 %. Species-specific PCR was performed on 58 cell culture samples where the presence of Mollicutes was previously detected. As a result, Mycoplasma orale was detected as the most frequent species, followed by Mycoplasma fermentans , Acholeplasma laidlawii , Mycoplasma arginini , Mycoplasma hyorhinis , and Mycoplasma salivarium . The results of this study confirm that the species-specific PCR assays developed are feasible to use for the identification of mycoplasma species in cell cultures.

Mycoplasmataceae

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Hallazgos histopatológicos en gallinas ponedoras afectadas por el virus de la bronquitis infecciosa aviar

Biotecnología aplicada (2012)

Manuel Colás Ana María Acevedo Alejandro Merino María del Carmen Lamazares Yaima Burgher

Histopathological fi ndings in egg-laying hens infected with avian infectious bronchitis virus. In order to dissect the histopathological changes produced by the infection of avian infectious bronchitis virus in previously vaccinated egg-laying hens from a poultry farming unit, 35 White Leghorn egg-laying hens that had been in pro- duction for 9 to 10 months (twenty seven of which had clinical symptoms corresponding to respiratory disease and eight apparently healthy individuals) were selected for further study. After clinical examination and necropsy, they were classifi ed into apparently healthy, mild, moderate or severe according to the severity of the clinical-pathological process. Samples were taken from paranasal sinuses, trachea and lungs for histopathological study, and trachea- lung pools were prepared from four individuals for virus isolation and molecular biology assays. The presence of mucus was evidenced with Schiff's non-enzymatic histochemical staining, and histomorphometric analyses were used to estimate the number of glands in the tracheal mucosa. The proportions of histopathological lesions were compared, using one-way Anova to determine gland loss at the tracheae with a signifi cance level of p < 0.05 in both cases. Histopathological analysis of the epithelia of paranasal sinuses, trachea and bronchia revealed the presence of epithelial erosion, mucous exudate and hyperplasia of mucosa-associated lymphoid tissue. Glandular cysts were observed at the paranasal sinuses, and epithelial metaplasia was detected in the trachea. It was possible to isolate and identify infectious bronchitis coronavirus from the original samples and from samples passaged in chicken embryos.

Histopathology

Mucous membrane

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Development of a PCR system with internal control for the reliable detection of mycoplasmas contaminating cell cultures and biological products

Revista de salud animal (2013)

Yaima Burgher

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Porcine Circovirus Modulates Swine Influenza Virus Replication in Pig Tracheal Epithelial Cells and Porcine Alveolar Macrophages

Yaima Burgher Chantale Provost Marie-Jeanne Pesant Carl A. Gagnon

The pathogenesis of porcine circovirus type 2b (PCV2b) and swine influenza A virus (SwIV) during co-infection in swine respiratory cells is poorly understood. To elucidate the impact of PCV2b/SwIV co-infection, newborn porcine tracheal epithelial cells (NPTr) and immortalized porcine alveolar macrophages (iPAM 3D4/21) were co-infected with PCV2b and SwIV (H1N1 or H3N2 genotype). Viral replication, cell viability and cytokine mRNA expression were determined and compared between single-infected and co-infected cells. Finally, 3’mRNA sequencing was performed to identify the modulation of gene expression and cellular pathways in co-infected cells. It was found that PCV2b significantly decreased and improved SwIV replication, in co-infected NPTr and iPAM 3D4/21 cells respectively, compared to single infected cells. Interestingly, PCV2b/SwIV co-infection synergistically up-regulated IFN expression in NPTr cells whereas in iPAM 3D4/21 cells, PCV2b impaired the SwIV IFN induced response, both correlating with SwIV replication modulation. RNA-sequencing analyses revealed that the modulation of gene expression and enriched cellular pathways during PCV2b/SwIV H1N1 co-infection is regulated in a cell type-dependent-manner. This study revealed different outcomes of PCV2b/SwIV co-infection in porcine epithelial cells and macrophages and provides new insights on porcine viral co-infections pathogenesis.

Pathogenesis

Circovirus

10.20944/preprints202304.0781.v1

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Experiencia de MYCOLAB en las actividades de Calidad y Acreditación como laboratorio de alcance microbiológico

REDVET. Revista Electrónica de Veterinaria (2013)

Yoesley Lozada Yaima Burgher Arianna Duque Arsenio Betancourt Armando Vega

Resumen es: Los micoplasmas son contaminantes frecuentemente de cultivos celulares, sueros y productos biofarmeceuticos de aplicacion medica, lo que hace necesario c...

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Validación del método de cultivo microbiológico para su utilización en el diagnóstico de micoplasmas en sueros, cultivos celulares y productos biofarmacéuticos

REDVET. Revista Electrónica de Veterinaria (2012)

Yoesley Lozada Evelyn Lobo Yaima Burgher Arianna Duque Siomara Martínez

Mycoplasmas frequently infected cell cultures, serums and biofarmeceutical products. With the development of biotechnology in Cuba, it is necessary to strict...

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Development of species-specific PCRs for the detection of mycoplasmas cell culture contaminants.

Revista de salud animal (2020)

Anisleidy Pérez Castillo Arianna Duque Ortiz Yaima Burgher Jose Antonio Agüero Fernández Mario Sergio Solis Castells

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