Current Situation of the Presence of Dirofilaria immitis in Dogs and Humans in Bucaramanga, Colombia.

The cardiopulmonary dirofilariosis caused by Dirofilaria immitis, is a vector-borne infection, which can be transmitted to humans. The main hosts are both domestic and wild canids. This species mainly occurs in tropical and subtropical climates, and temperature and humidity are the main factors that favor the presence and proliferation of culicid mosquitoes as vectors of the disease. There are few reports of cardiopulmonary dirofilariosis in dogs and humans in Colombia, a region with favorable climatic conditions which favors the presence of mosquitoes that act as vectors of the disease. Therefore, this study aimed to examine its current prevalence in dogs and the risk of human exposure to the disease in Bucaramanga, one of the most populated areas in Colombia located at the center of the country. Furthermore, its demographic and environmental characteristics could be useful as a study model for other similar locations and neighboring countries. Serum samples from 351 dogs and 506 humans from the Bucaramanga Metropolitan area were analyzed. All dog samples were analyzed by Knott's technique and tested with a commercial immunochromatographic to detect the presence of circulating antigens of D. immitis. Human samples were analyzed using a non-commercial ELISA test kit to detect IgG against the somatic antigens of adult D. immitis and Wolbachia. Positive results were further confirmed using western blot analysis. Thirty-eight dogs tested positive with a overall prevalence of 10.82%. Of these dogs, 18 showed D. immitis microfilariae, being 5.12% of the total population. The overall seroprevalence in humans was 6.71%; seroprevalence was significantly higher in individuals aged 16-34 years-old and in women than in men. To our knowledge, this study describes seropositivity to D. immitis for the first time in a Colombian human population located in the same area as that of dogs infected with D. immitis, which represents a potential threat to public health. In humans, age and gender can be considered risk factors for exposure to D. immitis.

Dengue in Santander State, Colombia: fluctuations in the prevalence of virus serotypes are linked to dengue incidence and genetic diversity of the circulating viruses.

OBJECTIVE: To investigate the link between fluctuations in the prevalence of dengue virus (DENV) serotypes and the number of dengue cases in the metropolitan area of Bucaramanga, Santander State, Colombia, in the 2007-2010 and 2014-2017 periods. METHOD: Viruses were isolated from febrile patient samples by direct application to C6/36-HT cells and typed using monoclonal antibodies. We performed autocorrelation and cross-correlation analyses to determine whether fluctuations in the prevalence of DENV serotypes and dengue cases were correlated. Full envelope (E) gene sequences were employed to examine the genetic diversity of serotypes circulating by using a phylogenetic approach. RESULTS: All four dengue virus serotypes were detected. DENV-1 was the dominant serotype in both periods followed by DENV-3 or DENV-2 depending on the period; DENV-4 was the least prevalent virus in both periods. Cross-correlation analyses suggest a temporal relation between the fluctuations in the prevalence of DENV serotypes, which were almost simultaneous (lag = 0) or related to recent past fluctuations (lag > 1.0) in the number of dengue cases. Data suggest that a sustained predominance of DENV-1, an increase of the DENV-4 prevalence, and a switch from DENV-3 to DENV-2 could be linked to an outbreak. Circulating viruses were grouped into Genotype V, Asia/American III and II for DENV-1, -2, -3 and -4, respectively; intragenotypic diversity was detected. CONCLUSIONS: The present work highlights the need of comprehensive studies on dynamics of DENV in Colombia to understand transmission of dengue and evaluate the effectiveness of a vaccination programme.

OBJECTIF: Etudier le lien entre les fluctuations de la prévalence des sérotypes du virus de la dengue (DENV) et le nombre de cas de dengue dans la région métropolitaine de Bucaramanga, dans l'Etat de Santander, en Colombie, au cours des périodes 2007-2010 et 2014-2017. MÉTHODE: Les virus ont été isolés à partir d'échantillons de patients fébriles par application directe sur des cellules C6/36-HT et typés à l'aide d'anticorps monoclonaux. Nous avons effectué des analyses d'autocorrélation et de corrélation croisée afin de déterminer si les fluctuations de la prévalence des sérotypes du DENV et des cas de dengue étaient corrélées. Des séquences de gènes d'enveloppe complète (E) ont été utilisées pour examiner la diversité génétique des sérotypes en circulation en utilisant une approche phylogénétique. RÉSULTATS: Tous les quatre sérotypes du virus de la dengue ont été détectés. DENV-1 était le sérotype dominant dans les deux périodes, suivi de DENV-3 ou DENV-2, selon la période; le virus DENV-4 était le moins prévalent au cours des deux périodes. Les analyses de corrélation croisée suggèrent une relation temporelle entre les fluctuations de la prévalence des sérotypes de DENV, qui étaient presque simultanées (lag = 0) ou liées aux fluctuations passées récentes (lag > 1,0) du nombre de cas de dengue. Les données suggèrent qu'une prédominance durable de DENV-1, qu'une augmentation de la prévalence de DENV-4 et qu'un passage de DENV-3 à DENV-2 pourraient être liés à une éclosion. Les virus en circulation ont été regroupés dans les génotypes V, Asie/Amérique III et II pour DENV-1, -2, -3 et -4, respectivement; une diversité intra-génotypique a été détectée. CONCLUSIONS: Le présent travail souligne la nécessité d'études approfondies sur la dynamique du DENV en Colombie afin de comprendre la transmission de la dengue et évaluer l'efficacité d'un programme de vaccination.

Discovery of antiviral molecules for dengue: In silico search and biological evaluation.

BACKGROUND: Dengue disease is a global disease that has no effective treatment. The dengue virus (DENV) NS2B/NS3 protease complex is a target for designing specific antivirals due to its importance in viral replication and its high degree of conservation. METHODS: NS2B/NS3 protease complex structural information was employed to find small molecules that are capable of inhibiting the activity of the enzyme complex. This inhibitory activity was probed with in vitro assays using a fluorescent substrate and the complex NS2B/NS3 obtained by recombinant DNA techniques. HepG2 cells infected with dengue virus serotype 2 were used to test the activity against dengue virus replication. RESULTS: A total of 210,903 small molecules from PubChem were docked in silico to the NS2B/NS3 structure (PDB: 2FOM) to find molecules that were capable of inhibiting this protein complex. Five of the best 500 leading compounds, according to their affinity values (-11.6 and -13.5 kcal/mol), were purchased. The inhibitory protease activity on the recombinant protein and antiviral assays was tested. CONCLUSIONS: Chemicals CID 54681617, CID 54692801 and CID 54715399 were strong inhibitors of NS2B/NS3, with IC50 values (µM) and percentages of viral titer reductions of 19.9, 79.9%; 17.5, 69.8%; and 9.1, 73.9%, respectively. Multivariate methods applied to the molecular descriptors showed two compounds that were structurally different from other DENV inhibitors. GENERAL SIGNIFICANCE: This discovery opens new possibilities for obtaining drug candidates against Dengue virus.