First Molecular Detection and Epidemiological Analysis of Equine Influenza Virus in Two Regions of Colombia, 2020-2023.

Equine influenza is a viral disease caused by the equine influenza virus (EIV), and according to the WOAH, it is mandatory to report these infections. In Latin America and Colombia, EIV risk factors have not been analyzed. The objective of this research is to perform an epidemiological and molecular analysis of the EIV in horses with respiratory symptoms from 2020 to 2023 in Colombia. Molecular EIV detection was performed using RT-qPCR and nanopore sequencing. A risk analysis was also performed via the GEE method. A total of 188 equines with EIV respiratory symptoms were recruited. The positivity rate was 33.5%. The descriptive analysis showed that only 12.8% of the horses were vaccinated, and measures such as the quarantine and isolation of symptomatic animals accounted for 91.5% and 88.8%, respectively. The variables associated with the EIV were the non-isolation of positive individuals (OR = 8.16, 95% CI (1.52-43.67), p = 0.014) and sharing space with poultry (OR = 2.16, 95% CI (1.09-4.26), p = 0.027). In conclusion, this is the first EIV investigation in symptomatic horses in Colombia, highlighting the presence of the virus in the country and the need to improve preventive and control measures.

Phylogenetic analysis of Hepatitis E virus strains isolated from slaughter-age pigs in Colombia.

Hepatitis E virus (HEV) is a zoonotic virus that causes acute hepatitis in humans, and can be transmitted via the fecal-oral route. Pigs are considered to be a reservoir for this infection-mainly where the disease is not endemic. In a previous study conducted in Antioquia, which is a region in Colombia where the production and consumption of pork meat is higher than in the rest of the country, the presence of anti-HEV IgG-type antibodies was reported in slaughter-age pigs. Aiming to identify the HEV genotype circulating in swine, animal liver, and feces samples from five swine cattle slaughterhouses located in six different sub-regions of Antioquia were collected. A nested RT-PCR (nRT-PCR) was used in order to amplify the HEV ORF-1 (170bp) and ORF-2 (348, and 958bp). The amplicons yielded in this study were sequenced, and a molecular phylogeny analysis based on the maximum likelihood method, including HEV sequences reported in several countries, was performed. Phylogeny analysis revealed that HEV amplification fragments from Antioquia's pigs were grouped in three clades within the sub-genotype 3a without a specific geographical structure, and were also genetically related to Japanese and American HEV sequences. This analysis provides the first approach on the genetic diversity and circulation dynamics of HEV in Colombian herds.

Cytotoxicity and in vitro activity of chard (Beta vulgaris L. var Cicla) extracts on porcine pancreatic islets / Citotoxicidad y actividad in vitro de extractos de acelga (Beta vulgaris L. var Cicla) en islotes pacreáticos porcinosa / Citotocidae e atividade in vitro de extratos de acelga (Beta vulgaris L. var Cicla) em ilhotas pancreáticas porcinas

Background: reports from traditional medicine suggest that chard (Beta vulgaris L. var Cicla) can have remarkable effects in diabetes therapy. Objective: to evaluate the cytotoxic activity of chard extracts in cell lines and determine the viability of cultured porcine pancreatic islets added with or without chard extracts. Methods: cytotoxic activity of chard extracts was assessed in non-tumor and tumor cell lines using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] technique, and the ability of extracts to maintain porcine pancreatic islets viability and regeneration in vitro was tested. Results: the 50% cytotoxic concentration (CC50) of extracts for non-tumor cell lines was above 1,000 μg/mL, while it was 825 μg/mL, 283 μg/mL, 136 μg/mL and 380 μg/mL, for hexane, ethyl acetate, ethanol and water extracts in the tumor cell line, respectively. The CC50 ratio between cell lines indicates that ethanol extract is 7.5 times more toxic to tumor than non-tumor cell lines. There was an increase in viability of porcine pancreatic islets cultured with aqueous, ethyl acetate, and ethanol extracts compared with standard media (CMRL1066) and Cyclosporine A (CsA) control groups. Furthermore, a greater than one regeneration index of islets cultured with ethanol extract at 1,000 μg/mL and 500 μg/mL concentrations during 15 days was observed, which remained constant and was significantly higher than CsA group. Conclusions: these results suggest that chard metabolites should be researched to develop antitumor therapies and human pancreatic islets recovery in diabetes treatment.

Antecedentes: reportes de medicina tradicional sugieren que la planta acelga (Beta vulgaris L. var Cicla) es importante en el tratamiento de enfermedades como la diabetes. Objetivo: evaluar la citotoxicidad de concentraciones de extractos de acelga en líneas celulares y determinar la viabilidad de islotes pancreáticos porcinos cultivados con y sin extracto de acelga. Método: se evaluó la actividad citotóxica en líneas celulares tumorales y no tumorales, con la técnica del MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide]. Específicamente se hicieron ensayos para comprobar si los extractos de acelga tienen la capacidad de mantener la viabilidad de islotes pancreáticos porcinos aislados e influir en su regeneración in vitro. Resultados: la concentración citotóxica al 50% (CC50) de los extractos en líneas no tumorales fue mayor de 1.000 μg/mL, mientras que para los extractos en hexano, acetato de etilo, etanol y agua fue de 825 μg/mL, 283 μg/mL, 136 μg/mL y 380 μg/mL, respectivamente, en líneas tumorales. La proporción CC50 encontrada indica que el extracto en etanol es 7,5 veces más tóxico para las líneas celulares tumorales que para las no tumorales. Igualmente encontramos un aumento en la viabilidad de los islotes pancreáticos porcinos cultivados con extracto acuoso, de acetato en etilo y etanol en comparación con el medio de cultivo estándar (CMRL1066) y un control inhibidor que contenía medio con Ciclosporina A (CsA). Además, se encontró que el índice de regeneración era mayor de uno en los islotes cultivados con el extracto en etanol a concentraciones de 1.000 μg/mL y 500 μg/mL durante 15 días, que se mantuvo constante y fue significativamente mayor en comparación con el grupo de CsA. Conclusión: estos resultados sugieren que los metabolitos de la acelga podrían ser utilizados en la investigación de nuevos fármacos para el desarrollo de terapias antitumorales y recuperación de islotes pancreáticos en el tratamiento de la diabetes.

Antecedentes: relatos encontrados em medicina sugerem que a planta acelga (Beta vulgaris L. var Cicla) tem un papel importante no tratamento das doenças como a diabetes. Objetivo: avaliar a citotoxicidade de concentrações de extratos em linhagens celulares e determinar a viabilidade de ilhotas pancreáticas de porcos cultivadas com e sem extrato de acelga. Métodos: neste trabalho foi avaliada a atividade citotóxica dos extratos da acelga em linhagens celulares tumorais e não tumorais, usando a técnica do MTT [3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide]; além disso, foram feitos ensaios para verificar a capacidade que têm os extratos para manter a viabilidade das ilhotas pancreáticas isoladas de porcos e a influência em sua regeneração in vitro. Resultados: a concentração citotóxica ao 50% (CC50) dos extratos em linhagens não tumorais está acima de 1000 μg/mL, enquanto para os extratos de hexano, acetato de etilo, etanol y água é de 825 μg/mL, 283 μg/mL, 136 μg/mL y 380 μg/mL, respectivamente, em linhagens tumorais. A proporção CC50 entre a célula indica que o extrato de etanol é 7,5 vezes mais tóxico para as linhas celulares tumorais que para as linhas não tumorais. Houve um aumento na viabilidade dos isolados pancreáticos de porcos cultivados com extrato aquoso, de acetato de etilo y etanol, em comparação com o meio de cultura padrão (CMRL 1066) e um controle inibitório contendo meio com Ciclosporina A (CsA). Encontrou-se também uma taxa de regeneração maior do que um em ilhotas cultivadas com concentrações de 1000 μg/mL e 500 μg/mL durante 15 días, que se manteve constante e foi significativamente mais elevada em comparação com a CsA. Conclusões: estes resultados sugerem que os metabolitos da acelga poderiam ser usados para a pesquisa de novas drogas para o desenvolvimento de terapias antitumorais e recuperação de ilhotas pancreáticas para o tratamento da diabetes.

Actividad antiviral in vitro de extractos de Hura crepitans y Codiaeum variegatum en la replicación de herpes virus bovino tipo-1 y virus de estomatitis vesicular / In vitro antiviral activity of extracts from Hura crepitans y Codiaeum variegatum on bovine herpes virus type-1 and vesicular stomatitis virus replication

Hojas de las plantas Hura crepitans y Codiaeum variegatum se recolectaron en la ciudad de Medellín (Colombia), para determinar la actividad biológica (Actividad citotóxica [AC] y antiviral [AAV]) de cuatro extractos de éstas (hexánico, en acetato de etilo, metanólico y acuoso) sobre los virus de Estomatitis Vesicular (VSV) serotipos Indiana y New Jersey y Herpes Virus Bovino tipo-1 cepa Bogotá (BHV-1B). El tamizaje de actividad biológica de los extractos se realizó en monocapas de células BHK-21. Luego los extractos promisorios, pasaron a la fase cuantitativa de AAV por el ensayo de MTT [3-(4,5-dimethylthiazol–2-yl)-2,5-diphenyltetrazolium bromide] para determinar la concentración citotóxica 50 (CC50), la concentración inhibitoria 50 (IC50) y el índice de selectividad (IS). Ninguno de los extractos de las cuatro plantas presentó AAV contra VSV, mientras que los extractos hexánico, en acetato de etilo y metanólico de H. crepitans confirieron resistencia a la infección por BHV-1B con IC50 de 17.41, 7.87 y 2.75 µ g/ml, respectivamente. El extracto con mejor IS (relación entre IC50 y CC50) fue el hexánico de H. crepitans (IS > 17.4). Estos resultados sugieren que pueden existir compuestos en el extracto hexánico de H. crepitans con una actividad promisoria anti BHV-1B. Este estudio es pionero en demostrar actividad antiviral de extractos de plantas de la especie Euphorbiaceae contra BHV-1B.

Leaves from Hura crepitans and Codiaeum variegatum plants were collected in Medellín, Colombia to determine its cytotoxic [CA] and antiviral activity [AVA]. Four extracts were prepared using hexane, ethyl acetate, methanol and water; the antiviral activity was tested against Vesicular Stomatitis Virus (VSV), serotypes Indiana and New Jersey and Bovine Herpes Virus (type 1 Bogotá strain). The preliminary biological activity of each of the eight extracts was determined on BHK-21 cell monolayers using 96 well-plates. Then, promissory extracts were further tested using the MTT [3-(4,5-dimethylthiazol–2-yl)-2,5-diphenyltetrazolium bromide] assay to determine the cytotoxic concentration 50 (CC50), the inhibitory concentration 50 (IC50 ), and the selectivity index (SI). None of the evaluated plants exhibited AVA against VSV, whereas the extracts in hexane, ethyl acetate and methanol from Hura crepitans protected cell monolayer from infection against BHV-Bogotá with IC50 of 17.41, 7.87 and 2.75 µ g/ml respectively. The best IS was hexane of H. crepitans (SI > 17.4). These results suggest that this extract contains antiviral compounds. This research is pioneer demonstrating AVA against BHV-1B in extracts from plants of the Euphorbiaceae family.