Anisleidy Pérez Castillo

Universidade Federal de Minas Gerais

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Evelyn Lobo Rivero

Centro Nacional de Sanidad Agropecuaria

Arianna Duque Ortiz

Centro Nacional de Sanidad Agropecuaria

Yaima Burgher

Université de Montréal

Evelyn Lobo-Rivero

Centro Nacional de Sanidad Agropecuaria

Júlia Angélica Gonçalves da Silveira

Universidade Federal de Minas Gerais

Mario Sergio Solis Castells

Centro Nacional de Sanidad Agropecuaria

Marina L. Meli

University of Zurich

Regina Hofmann‐Lehmann

University of Zurich

Renato Santana Aguiar

Universidade Federal de Minas Gerais

Arsenio Betancourt Bravo

Centro Nacional de Sanidad Agropecuaria

Cooperative Institutions

Universidade Federal de Minas Gerais

Centro Nacional de Sanidad Agropecuaria

Universidade de São Paulo

Universidade Federal do Paraná

Purdue University West Lafayette

Universidade Federal do Rio de Janeiro

ORCID

Universidade Estadual de Santa Cruz

University of Zurich

Instituto Federal de Educação, Ciência e Tecnologia de Minas Gerais

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Criopreservación y almacenamiento de Mycoplasma spp.

Revista de salud animal (2016)

Anisleidy Pérez Castillo Arsenio Betancourt Bravo Arianna Duque Ortiz Evelyn Lobo Rivero

Cryopreservation has as a principle to maintain cell viability and functionality at low temperatures; thus, to understand and properly implement cryopreservation of biological material is essential in laboratories and cell banks. In the case of mycoplasmas, maintenance of viable cultures is not only an operational issue, but also an essential factor in the good performance of the laboratory work. Therefore, this study aimed to assess the viability of mycoplasma strains preserved by freezing, in different matrices and several cryopreserving agents. For the study, it was used a culture of Mycoplasma hyorhinis (selected as type strain) with 5,1x108UFC/ml, which was inoculated into cell culture matrices and ascetic liquid. Both types of samples were stored at -70oC and -20oC by using or not cryopreserving agents; in this case, 5% glycerol and 2.5% dimethylsulfoxide were used. At 3, 30, 60 and 90 days postfreezing, mycoplasma viability was checked by growing them in liquid and solid Hayflick's media. As a result, it was found that in the matrices frozen at -20oC and -70oC and cryopreserving, Mycoplasma hyorhinis remained viable for 90 days, but not in the matrices containing no cryopreserving, where the behavior was variable depending on the temperature. It was concluded that the addition of cryopreserving agents (5% glycerol or 2.5% dimethylsulfoxide) preserved the viability of mycoplasmas in the matrices studied during freezing and subsequent storage; also the temperature of -20oC can be used by those laboratories lacking equipment with -70oC to store samples for three months, period of time designed for interlaboratory studies. Key words: mycoplasmas, cryopreservation, storage, quality, diagnosis.

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Seroprevalence of Anti-Sars-Cov-2 Antibodies in Cats and Dogs of Belo Horizonte, Minas Gerais, Brazil

Viviane Campos Spanhol Cláudia Fideles Resende Marina Mourão Sena Claudino Amanda Melo Ana Moura

Seroprevalence

2019-20 coronavirus outbreak

10.2139/ssrn.4921244

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SARS-CoV-2 surveillance in captive animals at the belo horizonte zoo, Minas Gerais, Brazil

Virology Journal (2024)

Anisleidy Pérez Castillo João Victor Oliveira Miranda Paula Luize Camargos Fonseca Rennan Garcias Moreira Luíza Campos Guerra de Araújo e Santos

The pandemic caused by SARS-CoV-2 has not only affected humans but also raised concerns about its transmission to wild animals, potentially creating natural reservoirs. Understanding these dynamics is critical for preventing future pandemics and developing control strategies. This study aims to investigate the presence of SARS-CoV-2 in wild mammals at the Belo Horizonte Zoo in Brazil, analyzing the virus's evolution and zoonotic potential. The study was conducted at the Belo Horizonte Zoo, Minas Gerais, Brazil, covering a diverse population of mammals. Oropharyngeal, rectal, and nasal swabs were collected from 47 captive animals between November 2021 and March 2023. SARS-CoV-2 presence was determined using RT-PCR, and positive samples were sequenced for phylogenetic analysis. Consensus genomes were classified using Pangolin and NextClade tools, and a maximum likelihood phylogeny was inferred using IQ-Tree. Of the 47 animals tested, nine (19.1%) were positive for SARS-CoV-2. Positive samples included rectal, oropharyngeal, and nasal swabs, with the highest positivity in rectal samples. Three genomes were successfully sequenced, revealing two variants: VOC Alpha in a maned wolf (Chrysocyon brachyurus) and a fallow deer (Dama dama), and VOC Omicron in a western lowland gorilla (Gorilla gorilla gorilla). Phylogenetic analysis indicated potential human-to-animal transmission, with animal genomes clustering close to human samples from the same region. This study highlights the presence of SARS-CoV-2 in various wild mammal species at the Belo Horizonte Zoo, emphasizing the virus's zoonotic potential and the complexity of interspecies transmission. The detection of different variants suggests ongoing viral evolution and adaptation in new hosts. Continuous monitoring and genomic surveillance of SARS-CoV-2 in wildlife are essential for understanding its transmission dynamics and preventing future zoonotic outbreaks. These findings underscore the need for integrated public health strategies that include wildlife monitoring to mitigate the risks posed by emerging infectious diseases.

Gorilla

Pandemic

Mammal

Natural reservoir

Zoonosis

10.1186/s12985-024-02505-9

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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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Molecular detection and characterization of Hepatozoon canis in stray dogs from Cuba

Parasitology International (2020)

Adrian Alberto Díaz-Sánchez Regina Hofmann‐Lehmann Marina L. Meli Lisset Roblejo‐Arias Osvaldo Fonseca‐Rodríguez

Hepatozoon

10.1016/j.parint.2020.102200

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Citations (17)

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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Molecular Diagnosis, Prevalence and Importance of Zoonotic Vector-Borne Pathogens in Cuban Shelter Dogs—A Preliminary Study

Pathogens (2020)

Adrian Alberto Díaz-Sánchez Belkis Corona Marina L. Meli Lisset Roblejo‐Arias Osvaldo Fonseca‐Rodríguez

The present study aimed to determine the prevalence of zoonotic vector-borne pathogens, including

Ehrlichia canis

Anaplasma phagocytophilum

Rhipicephalus sanguineus

Babesia canis

10.3390/pathogens9110901

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Citations (8)

Cryopreservation and storage of Mycoplasma spp.

Revista de salud animal (2016)

Anisleidy Pérez Castillo Arsenio Betancourt Bravo Arianna Duque Ortiz Evelyn Lobo Rivero

Mycoplasmataceae

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Molecular detection and characterization of SARS-CoV-2 in cats and dogs of positive owners during the first COVID-19 wave in Brazil

Scientific Reports (2023)

Juliana Arena Galhardo David Soeiro Barbosa Louise Bach Kmetiuk Otávio Carvalho Ana Izabel Passarella Teixeira

Despite previous reports of SARS-CoV-2 infection in dogs and cats worldwide, the type of swab sample used for its detection through RT-qPCR needs to be better compared and described. Accordingly, as part of a multicenter study in Brazil, the aim of the present study was to assess which rectal or oropharyngeal swabs would be more appropriate for detecting SARS-CoV-2 in cats and dogs, through viral load comparison. Pets of owners diagnosed with COVID-19 in the last 7 days were eligible. A total of 148 animals from four of the five Brazilian geographical regions were analyzed, among which 10/48 cats (20.83%) and 11/100 dogs (11.00%) were positive. The results suggested that oropharyngeal swabs should be considered for SARS-CoV-2 detection, particularly in cats, due to the higher cDNA viral load. Also, the genomic results showed similarities between SARS-CoV-2 animal variants and human variants that were circulating at the time of sampling, thus corroborating the existence of zooanthroponotic transmission. In conclusion, the present study highlighted the importance of SARS-CoV-2 monitoring among cats and dogs, as virus modification may indicate the possibility of mutations in animals and spillover back to owners. Thus, positive individuals should always self-isolate from their pets during COVID-19, to prevent trans-species transmission and mutation.

2019-20 coronavirus outbreak

10.1038/s41598-023-41285-0

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Citations (4)

Notes sobre canonges de la catedral de Barcelona i els seus testaments (segle XII)

Analecta sacra tarraconensia: Revista de ciències historicoeclesiàstiques (1998)

Anisleidy Pérez Castillo

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