FAA. Investigaciones Biomédicas

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An Approach to the Identification and Phylogenetic Analysis of Emerging and Hemorrhagic Fever Viruses.

Díaz, Francisco J. | 2019-11-22

An important aspect of virological surveillance is the identification of the detected viruses. Broad surveillance, that typically employs deep sequencing of collected tissue samples, provides the investigator with many sequence files constructed from overlapping stretches of DNA sequences. Directed surveillance for viruses of a specific taxonomic group provides the investigator with sequence files from cDNA amplified using specific primers to conserved viral regions. Here we will describe general approaches to identify hemorrhagic viral agents through phylogenetic analysis of cDNA sequences obtained during surveillance activities.

Sampling Design and Mosquito Trapping For Surveillance of Arboviral Activity.

Paternina, Luís E. | 2019-11-22

Mosquitoes are the most important vectors for arboviral human diseases across the world. Diseases such as Dengue Fever (DF), West Nile Virus (WNV), Yellow Fever (YF), Japanese Encephalitis (JE), Venezuelan Equine Encephalitis (VEE), and St. Louis Encephalitis (SLE), among others, have a deep impact in public health. Usually mosquitoes acquire the arboviral infection when they feed on viremic animals (birds or mammals), so their infection can be detected along the year or in short periods of time (seasons). All of this depends on the frequency and seasonality of the encounters between viremic animals and vectors. With the convergence of several phenomena like the increasing traveling of human populations, globalization of economy and more recently the global warming, the introduction of nonendemic arbovirus into new areas has become the current scenario. As examples of this new social and environmental frame we can mention the outbreak of West Nile Virus in North America in the late 1990s and more recently the outbreaks of chikungunya and Zika virus in the Americas. The present chapter deals with one of the first steps in the development of research studies and diagnosis programs, the surveillance of arboviruses in their vectors, the sampling design and mosquito trapping methods. The chapter also includes some important considerations and tips to be taken into account during the mosquito fieldwork.

Dengue del virus 1 aislado de la poliproteína Ov07, región de la proteína E de la envoltura, gen (POLY), CD parcial

Camacho Burgos, Erwin Yesid | 2019-04-10

Detection of Dengue Virus in Department of Bolivar, Colombia, 2012- 2013, Real-Time PCR.

Gómez Camargo, Doris Esther | 2019-11-12

To assess the presence of RNA from dengue virus by Real Time Polymerase Chain Reaction with Reverse Transcriptase (qRT-PCR), in anti-dengue IgM seronegative serum samples obtained from symptomatic patients with less than 5 days of fever, in the department of Bolivar, Colombia.

Seroprevalencia a Rickettsia del grupo de las fiebres manchadas, en población humana de zona rural del municipio de Toluviejo, Colombia.

Arrieta Hernández, Norma | 2019-11-12

Rickettsia is a genus consisting of obligate intracellular bacteria; the main vectors are arthropod ectoparasites, espe-cially ticks. In 2008, was detected in Toluviejo, Colombia, a seropositive person to Rickettsia, however, there is little information about the prevalence in this municipality.Objetives. To study the seroprevalence of Rickettsia of the spotted fever group and it’s relation with socio-demographic factors in a human population in Toluviejo, Colombia.

First report of Amblyomma dissimile (Acari: Ixodidae) on Spilotes pullatus (Squamata: Colubridae) from Colombia.

Verbel Vergara, Daniel E. | 2019-11-08

The very few studies about the tick fauna of Colombia have focused on ectoparasites of domestic and farm animals, therefore little information about the tick fauna in wildlife is available. The purpose of this research is to contribute to the knowledge of ticks that parasitize snakes in Colombia.

Comparación y utilidad de tres métodos de extracción de ADN a partir de potenciales vectores de enfermedades rickettsiales (Acari: Ixodidae) en Colombia.

Guevara Vega, Marco | 2019-11-08

The study of diseases transmitted for tick as rickettsia is of great interest to medical and veterinary level and requires the use of extracts of good quality DNA for subsequent analysis of amplified genes of these pathogens. In this study, three methods were compared DNA extraction from ticks complex Rhipicephalus sanguineus based on: chaotropic guanidine thiocyanate (method 1), high salt (method 2) and an extraction method based on commercially available columns (Method 3). The DNA was quantified and the quality of each extract in the amplification of the 16S gene ticks (360pb) was evaluated, a nonparametric Kruskal-Wallis one-way analysis was performed, while comparing the amplification efficiency of mitochondrial gene was performed by Dunn test range. Method 3 allowed the amplification of the 16S gene of ticks in 100% (30/30) followed by method 1 with 93.33% (28/30) and finally with method 2 86.66% (26/30). The three methods are efficient at extracting genetic material of high quality tick-borne studies since their performance in amplification of mitochondrial genes tick agents.

Distribución geográfica de especies de garrapatas infectadas con Babesia de zonas rurales del departamento de Sucre, Colombia.

Guevara Vega, Marco | 2019-11-08

The agents of the genus Babesia (Piroplasma: Babesiidae) are hemoparasites protozoa transmitted by ticks of the family Ixodidae during blood meal. In Colombia, the few studies of ticks, have focused on aspects of pest control in veterinary medicine and its impact on livestock, with very few contributions to the study of these ectoparasites as potential vectors of zoonotic diseases. In this paper, distribution models for tick species as R. (Boophilus) microplus, D. nitens, and R. sanguineus to estimate risk areas ixodológico in the Caribbean region were created. This distribution modeling was built by algorithms such as Maximum Entropy, and implemented in 3.3.3k Maxent program, taking the geographical points where there has been the presence of tick species and environmental variables to describe the ecological space used by each of them. All projections geographic distribution models generated were processed in QGIS 2.8.2 program. It was obtained as a result that some areas of the department of Cordoba can probably be these tick species and absent in places like the Sierra Nevada de Santa Marta. This is the first step in determining which areas are likely to find potential vectors of zoonotic agents of the genus Babesia.

Molecular Evidence Confirms the Taxonomic Separation of Lutzomyia tihuiliensis from Lutzomyia pia (Diptera: Psychodidae) and the Usefulness of Pleural Pigmentation Patterns in Species Identification.

Pérez Doria, Alveiro | 2019-11-08

The phlebotomine sand flies Lutzomyia pia (Fairchild & Hertig 1961) and Lutzomyia tihuiliensis Le Pont, Torrez-Espejo & Dujardin 1997 (Diptera: Psychodidae) belong to the pia series of the Lu. verrucarum species group, which includes several species that bite humans in Andean foci of leishmaniasis. The females of these two species exhibit isometry and isomorphism in anatomical structures of the head and terminalia commonly used in taxonomic identipcation of sand bies. They can only be differentiated based on subtle differences in the pigmentation of the pleura. In Lu. tihuiliensis, this is restricted to the basal portions of the katepimeron and katepisternum, whereas in Lu. piaboth structures are totally pigmented. Takinginto account the subtle morphological differences between these species, the objective of the current study was to evaluate the specipc taxonomic status of Lu. tihuiliensis with respect to Lu. pia. A 475-bp portion of the mitochondrial genome was sequenced, composed of the 3 end of the cytochrome b gene, intergenic spacer 1, the transfer RNA gene for serine, intergenic spacer 2, and the 3 end of the gene NAD dehydrogenase 1. Genetic analysis conÞrms that Lu. tihuiliensis and Lu. pia constitute two distinct species and this is supported by four strong lines of evidence, i.e., the paired genetic distances, size differences and amino acid composition of the cytochrome b protein, presence and absence of intergenic spacer one and divergence observed in the sequence of the transfer RNA gene for serine. It also conÞrms the validity of the pleural pigmentation pattern as a species diagnostic character and the importance of performing a detailed examination of this character during morphological determination of phlebotomine sand flies in the series pia.

Presencia de Lutzomyia dubitans (Sherlock, 1962) (Diptera: Psychodidae: Phlebotominae) en la Ciudad de Sincelejo, Departamento de Sucre, Colombia.

Pérez Doria, Alveiro | 2019-11-07

El objetivo de esta nota es informar del hallazgo de Lutzomyia dubitans (Sherlock, 1962), en zona urbana de la ciudad de Sincelejo, Departamento de Sucre, Colombia. Los flebotomíneos fueron colectados con trampas de luz tipo CDC instaladas en el peridomicilio. Este hallazgo eleva a ocho el número de especies de Lutzomyia registradas en la ciudad, lo que pone en evidencia la tendencia hacia la colonización de áreas urbanas por los insectos transmisores de leishmaniosis.

Primer registro de Lutzomyia scorzai y Lutzomyia reburra (Diptera: Psychodidae) en Antioquia, Colombia.

Vergara, Daniela | 2019-10-07

Seven species of Lutzomyia and one of Brumptomyia were collected while surveying two cutaneous leishmaniosis foci in the department Antioquia, Colombia, these are: Lutzomyia gomezi, Lu. scorzai, Lu. trapidoi, Lu. triramula, Lu. panamensis, Lu. aragaoi, Lu. reburra and Brumptomyia galindoi. Lu. scorzai (subgenus Helcocyrtomyia) and Lu. reburra (subgenus Trichophoromyia) are new records for the department. The specimens of Lu. scorzai were captured using a Shannon light trap in the municipality of Tarazá, while those of Lu. reburra were collected using CDC-like light traps near a nest of armadillo in the municipality of Urrao. The biogeographic importance of these findings is discussed given the limited distribution of both species in Colombia.

Evidence for anthropophily in five species of phlebotomine sand flies (Diptera: Psychodidae) from northern Colombia, revealed by molecular identification of bloodmeals.

Paternina, Luís E. | 2019-11-06

Identification of the bloodmeal sources of phlebotomine sand flies is fundamental to determining which species are anthropophilic and understanding the transmission of Leishmania parasites in natural epidemiological settings. The objective of this study was to identify sand fly bloodmeals in the mixed leishmaniasis focus of the department of Sucre, northern Colombia. In all 141 engorged female sand flies were analyzed, after being captured in intradomiciliary, peridomiciliary and extradomiciliary habitats with Shannon and CDC traps and by active searching in diurnal resting sites. Bloodmeals were identified by sequencing and analysis of a 358 bp fragment of the mitochondrial gene Cytochrome b (CYB) and a 330 bp fragment of the nuclear gene prepronociceptin (PNOC). Using both genes 105 vertebrate bloodmeals were identified, with an efficiency of 72% for CYB but only 7% for PNOC. Ten species of vertebrates were identified as providing bloodmeal sources for eight sand fly species: Homo sapiens (Lutzomyia evansi, Lu. panamensis, Lu. micropyga, Lu. shannoni and Lu. atroclavata), Equus caballus (Lu. evansi, Lu. panamensis and Lu. cayennensis cayennensis), Eq. asinus (Lu. evansi and Lu. panamensis), Bos taurus 4 (Lu. evansi, Lu. panamensis and Lu. c. cayennensis), Tamandua mexicana (Lu. shannoni and Lu. trinidadensis), Proechimys guyanensis (Lu. evansi, Lu. panamensis and Lu. c. cayennensis), Mabuya sp. (Lu. micropyga), Sus scrofa (Lu. evansi and Lu. gomezi) and Gallus gallus (Lu. evansi). Cattle, donkeys, humans and pigs were significantly more important than other animals (P = 0.0001) as hosts of Lu. evansi, this being the most abundant sand fly species. The five Lutzomyia species in which blood samples of human origin were detected included Lu. micropyga and Lu. atroclavata, constituting the first evidence of anthropophily in both species.

Diagnostic accuracy of rKLO8 versus rK26 ELISAs for screening of canine visceral leishmaniasis.

Martínez Abad, Lily P. | 2019-11-05

Canine visceral leishmaniasis (CVL) represents an important public health issue. Despite numerous diagnostic tests available, CVL diagnosis still needs to be improved to achieve a more accurate detection rate. Recently, rKLO8, a new antigenic protein of Sudanese Leishmania donovani, was studied for the first time in diagnosis of human visceral leishmaniasis (HVL) and showed good performance. The present study aimed to evaluate serum reactivity to rKL08 and the reference antigen rK26, and to compare both diagnostic proteins with the combined DPP® CVL rapid test and ELISA (EIE-Bio-Manguinhos) confirmatory test, which are both recommended for the diagnosis of CVL in Brazil. Serum samples of dogs were grouped into: (I) DPP®/EIE negative (n=100) and (II) DPP®/EIE positive sera (n=100). Enhanced levels of IgG, mainly IgG2, to both rKLO8 and rK26 were found in group II. Sensitivity was 68% and 77% and specificity was 92% and 91%, for rKLO8 and rK26 antigens, respectively. Moreover, the combination of rKLO8 and rK26 antigens (rKLO8+rK26) exhibited higher sensitivity (85%) and specificity (93%). Thus, our results show that apart from the improved diagnostic power of rKLO8 in HVL, this new antigen is also suitable for the diagnosis of CVL. Further, the combination of rKLO8 and rK26 antigens increases the diagnostic accuracy of CVL.

Distribution of Spotted Fever Group Rickettsiae in Hard Ticks (Ixodida: Ixodidae) from Panamanian Urban and Rural Environments (2007-2013).

Bermúdez, Sergio E. | 2019-10-05

Tick-borne rickettsiosis is an important emerging disease in Panama; to date, there have been 12 confirmed cases, including eight fatalities. To evaluate the distribution of rickettsiae in Panamanian ticks, we collected questing and on-host ticks in urban and rural towns in elevations varying between 0 and 2300 m. A total of 63 sites (13 urban and 50 rural towns) were used to develop models of spatial distributions. We found the following tick species: Rhipicephalus sanguineus s.l. (present in 54 of 63 towns and cities), Amblyomma mixtum (45/63), Dermacentor nitens (40/63), A. ovale (37/63), Rhipicephalus microplus (33/63), A. oblongoguttatum (33/63), Ixodes affinis (3/63), and Ixodes boliviensis (2/63). Rhipicephalus sanguineus s.l. was present in urban and rural towns, and other species were present only in rural towns. DNA was extracted from 408 R. sanguineus s.l., 387 A. mixtum, 103 A. ovale, and 11 A. oblongoguttatum and later tested for rickettsiae genes using PCR. Rickettsia DNA was detected in ticks from 21 of 63 localities. Rickettsia rickettsii was detected in five A. mixtum (1.29%), and Candidatus ‘‘Rickettsia amblyommii’’ was found in 138 A. mixtum (35%), 14 R. sanguineus (3.4%), and one A. ovale (0.9%). These results suggest that much of rural Panama is suitable for the expansion of tick populations and could favor the appearance of new tick-borne rickettsiosis outbreaks.

L-Asparaginase from E. chrysanthemi expressed in glycoswitch: effect of Hist-Tag fusion on the extracellular expression.

Effer, Brian | 2019-10-31

L-Asparaginase (L-ASNase) is an important enzyme used to treat acute lymphoblastic leukemia, recombinantly produced in a prokaryotic expression system. Exploration of alternatives production systems like as extracellular expression in microorganisms generally recognized as safe (such as Pichia pastoris GlycoswitchVR ) could be advantageous, in particular, if this system is able to produce homogeneous glycosylation. Here, we evaluated extracellular expression into GlycoswitchVR using two different strains constructions containing the asnB gene coding for Erwinia chrysanthemi L-ASNase (with and without His-tag), in order to find the best system for producing the extracellular and biologically active protein. When the His-tag was absent, both cell expression and protein secretion processes were considerably improved. Three-dimensional modeling of the protein suggests that additional structures (His-tag) could adversely affect native conformation and folding from L-ASNase and therefore the expression and cell secretion of this enzyme.

Presence of Lutzomyia evansi, a vector of American visceral leishmaniasis, in an urban area of the Colombian Caribbean coast.

Bejarano, Eduar Elias | 2019-10-31

American visceral leishmaniasis (AVL) is a potentially fatal disease of neotropical countries, chiefly affecting children aged under 5 years, caused by ~ishmania (Leishmanial &a& Cunha & Chapas. which is indisiinguishablk from the ‘Old World’ paiasite Le. (Le.) injuntum Nicolle, of which it may be a synonym. AVL is transmitted to human or other mammalian hosts by the bites of the phlebotomine sandflies Lutzanzyia (Lutzomy&z) Zongt$alpis (Lutz & Neiva) or Lmmnyiu (Lutzomyia) evami (Nufiez-Tovar) (GRIMALDI et al., 1989; TRAVI et al., 1996). Although Lu. longipalpis is the most widespread and important vector of AVL, Lu. evarnsi has been confirmed as a primary vector in rural areas of the Caribbean coast of Colombia and in some areas of Venezuela (AGUIUR et aE., 1995; MONTOYA, 1996; FEIJCLWGELI et al., 19991. While conducting entomological studies in the urbad area of Sincelejo &y (9’ 18’ N. 75” 25’ WI in northern Colombia. we found adults of Lzk. eeransi ih houses and in peridbmiciliary environments. This is the first record of Lu. evansi from an urban area in Colombia.

Identification of rickettsia spp infection in patients with clinical suspicion of leptospirosis application of molecular techniques in a case series.

Gómez Carmago, Doris E. | 2019-10-30

Leptospirosis is a public health problem on the Colombian Caribbean Coast. In Cartagena, a major city in this region, this disease is endemic with high annual lethality. In clinical practice, differential diagnosis of leptospirosis with others icteric febrile syndromes is crucial to an appropriated therapeutic approach. Nevertheless, high complexity hospitals employ only serological test to confirm suspicious cases, with an elevated frequency of undetermined results. Therefore, the aim of this study was to apply molecular techniques in patients under leptospirosis suspicion. It was carried out a case series study with adults in 2013 last trimester. Patients with clinical manifestations and positive/undetermined results in serological test for leptospirosis were included. We used Polymerase Chain Reaction to detect Leptospira spp from blood samples with specific primers that amplify a DNA segment from the lipl32 gen. A total of 9 patients were included (3 women, 6 men). IgM test for leptospirosis was positive in 5 subjects and undetermined in 4 others. A 423pb product was expected, however only 3 samples showed an amplicon of 358pb. When those DNA fragments were sequenced, none of them were part of Leptospira spp. genome. Thereafter, samples were amplified for Riskettsia spp. by nested PCR for the OmpB gene. Seven samples were positive (420pb amplicon). These samples were also positive for Rickettsia spp 17KDa gene (230pb amplicon).

Detección parasitológica y molecular de Anaplasma spp en ganado bovino de Ovejas, Sucre (Colombia).

Carrillo Martínez, Auris C. | 2019-10-29

Uno de los obstáculos más relevantes de la ganadería nacional son las enfermedades transmitidas por ectoparásitos en particular la anaplasmosis bovina, una enfermedad causada por Anaplasma marginale. Esta infección se caracteriza por presentar amplia distribución mundial y por alcanzar especial importancia económica, debido a la disminución de la producción de carne y leche, mortalidad y pérdida de la eficiencia reproductiva por abortos. Hasta la fecha se desconoce la circulación del género Anaplasma en bovinos del municipio de Ovejas. El propósito de esta investigación es determinar la frecuencia de infección de Anaplasma spp en bovinos del municipio de Ovejas, Sucre (Colombia).

Zika virus like particles elicit protective antibodies in mice.

Salvo, Mauricio A | 2019-10-29

Mosquito-borne Zika virus (ZIKV) typically causes a mild and self-limiting illness known as Zika fever. Since its recent emergence in 2014 in the American continent, ZIKV infection during pregnancy has been closely associated with a wide range of congenital abnormalities. To date, no vaccines or antivirals are publicly available. We developed Zika virus-like particles (VLPs) and evaluated their immunogenicity and protective efficacy in mouse models. ZIKV VLPs (ZIKVLPs) formulated with alum were injected into 6-8-week-old interferon deficient AG129 mice as well as wild type BALB/c mice. Control mice received PBS/alum. Animals were challenged with 200 PFU (>1000 AG129 LD50s) of ZIKV strain H/PF/2013. All vaccinated mice survived with no morbidity or weight loss while control animals either died at 9 days post challenge (AG129) or had increased viremia (BALB/c). Neutralizing antibodies were observed in all ZIKVLP vaccinated mice. The role of neutralizing antibodies in protecting mice was demonstrated by passive transfer. Our findings demonstrate the protective efficacy of the ZIKVLP vaccine and highlight the important role that neutralizing antibodies play in protection against ZIKV infection.

Influencia de un programa de estimulación temprana en el desarrollo lingüístico de niños de Sincelejo.

Palacio Vásquez, Madelin María | 2019-10-29

There is evidence of the benefits of early stimulation on the development of abilities and abilities of children in biological or social risk condition, it is also known about the negative impact to the lack of stimulation on development. Objective: to determine the impact of an early stimulation program on language development in 3-year-old children in the municipality of Sincelejo.