Genetic similarities and phylogenetic analysis of Muntjac (Muntiacus spp. ) by comparing the nucleotide sequence of 16S rRNA and cytochrome B genome / Semelhanças genéticas e análise filogenética de muntjac (Muntiacus spp. ) comparando a sequência de nucleótidos de rRNA 16S e genoma citocromo B

This study aimed to identify the phylogenetic similarities among the muntjac (Muntiacus spp.). The phylogenetic similarities among seven major muntjac species were studied by comparing the nucleotide sequence of 16s rRNA and cytochrome b genome. Nucleotide sequences, retrieved from NCBI databases were aligned by using DNASTAR software. A phylogenetic tree was created for the selected species of muntjac by using the maximum likelihood method on MEGA7 software. The results of nucleotide sequences (16s rRNA) showed phylogenetic similarities between, the M. truongsonensis and M. rooseveltorum had the highest (99.2%) while the lowest similarities (96.8%) found between M. crinifrons and M. putaoensi. While the results of nucleotide sequences (Cty b) showed the highest similarity (100%) between M. muntjak and M. truongsonensis and the lowest s (91.5%) among M. putaoensis and M. crinifrons. The phylogenetic tree of muntjac species (16s rRNA gene) shows the main two clusters, the one including M. putaoensis, M. truongsonensis, M. rooseveltorum, and M. muntjak, and the second one including M. crinifrons and M. vuquangensis. The M. reevesi exists separately in the phylogenetic tree. The phylogenetic tree of muntjac species using cytochrome b genes shows that the M. muntjak and M. truongsonensis are clustered in the same group.

Este estudo visou identificar as semelhanças filogenéticas entre os muntjac (Muntiacus spp.). As semelhanças filogenéticas entre sete grandes espécies muntjac foram estudadas comparando a sequência de nucleótidos de 16s rRNA e genoma citocromo b. As sequências de nucleótidos, obtidas a partir de bases de dados NCBI, foram alinhadas utilizando o software DNASTAR. Foi criada uma árvore filogenética para as espécies selecionadas de muntjac utilizando o método de probabilidade máxima no software MEGA7. Os resultados das sequências de nucleótidos (16s rRNA) mostraram semelhanças filogenéticas entre o M. truongsonensis e o M. rooseveltorum tiveram o maior número (99,2%) enquanto as semelhanças mais baixas (96,8%) encontradas entre M. crinifrons e M. putaoensi. Enquanto os resultados das sequências de nucleótidos (Cty-b) apresentaram a maior semelhança (100%) entre M. muntjak e M. truongsonensis e os mais baixos (91,5%) entre M. putaoensis e M. crinifrons. A árvore filogenética das espécies muntjac (gene rRNA 16s) mostra os dois principais aglomerados, o que inclui M. putaoensis, M. truongsonensis, M. rooseveltorum e M. muntjak, e o segundo incluindo M. crinifrons e M. vuquangensis. O M. reevesi existe separadamente na árvore filogenética. A árvore filogenética das espécies muntjac usando genes citocromo b mostra que os M. muntjak e M. truongsonensis estão agrupados no mesmo grupo.

Genetic similarities and phylogenetic analysis of Muntjac (Muntiacus spp.) by comparing the nucleotide sequence of 16S rRNA and cytochrome B genome

Abstract This study aimed to identify the phylogenetic similarities among the muntjac (Muntiacus spp.). The phylogenetic similarities among seven major muntjac species were studied by comparing the nucleotide sequence of 16s rRNA and cytochrome b genome. Nucleotide sequences, retrieved from NCBI databases were aligned by using DNASTAR software. A phylogenetic tree was created for the selected species of muntjac by using the maximum likelihood method on MEGA7 software. The results of nucleotide sequences (16s rRNA) showed phylogenetic similarities between, the M. truongsonensis and M. rooseveltorum had the highest (99.2%) while the lowest similarities (96.8%) found between M. crinifrons and M. putaoensi. While the results of nucleotide sequences (Cty b) showed the highest similarity (100%) between M. muntjak and M. truongsonensis and the lowest s (91.5%) among M. putaoensis and M. crinifrons. The phylogenetic tree of muntjac species (16s rRNA gene) shows the main two clusters, the one including M. putaoensis, M. truongsonensis, M. rooseveltorum, and M. muntjak, and the second one including M. crinifrons and M. vuquangensis. The M. reevesi exists separately in the phylogenetic tree. The phylogenetic tree of muntjac species using cytochrome b genes shows that the M. muntjak and M. truongsonensis are clustered in the same group.

Resumo Este estudo visou identificar as semelhanças filogenéticas entre os muntjac (Muntiacus spp.). As semelhanças filogenéticas entre sete grandes espécies muntjac foram estudadas comparando a sequência de nucleótidos de 16s rRNA e genoma citocromo b. As sequências de nucleótidos, obtidas a partir de bases de dados NCBI, foram alinhadas utilizando o software DNASTAR. Foi criada uma árvore filogenética para as espécies selecionadas de muntjac utilizando o método de probabilidade máxima no software MEGA7. Os resultados das sequências de nucleótidos (16s rRNA) mostraram semelhanças filogenéticas entre o M. truongsonensis e o M. rooseveltorum tiveram o maior número (99,2%) enquanto as semelhanças mais baixas (96,8%) encontradas entre M. crinifrons e M. putaoensi. Enquanto os resultados das sequências de nucleótidos (Cty-b) apresentaram a maior semelhança (100%) entre M. muntjak e M. truongsonensis e os mais baixos (91,5%) entre M. putaoensis e M. crinifrons. A árvore filogenética das espécies muntjac (gene rRNA 16s) mostra os dois principais aglomerados, o que inclui M. putaoensis, M. truongsonensis, M. rooseveltorum e M. muntjak, e o segundo incluindo M. crinifrons e M. vuquangensis. O M. reevesi existe separadamente na árvore filogenética. A árvore filogenética das espécies muntjac usando genes citocromo b mostra que os M. muntjak e M. truongsonensis estão agrupados no mesmo grupo.

Infección natural por SARS-CoV-2 en gatos y perros domésticos de personas con diagnóstico de COVID-19 en el Valle de Aburrá, Antioquia / Natural SARS-CoV- 2 infection in domestic cats and dogs of humans diagnosed with COVID-19 in Valle de Aburrá, Antioquia

Introducción. El síndrome respiratorio agudo grave causado por el nuevo coronavirus SARS-CoV-2 es causa de la emergencia sanitaria por la pandemia de COVID-19. Si bien el humano es el principal huésped vulnerable, en estudios experimentales y reportes de infección natural, se han encontrado casos de zoonosis inversa de SARS-CoV-2 en animales. Objetivo. Evaluar la infección natural por SARS-CoV-2 en gatos y perros de propietarios con diagnóstico de COVID-19 en el Valle de Aburrá, Antioquia, Colombia. Materiales y métodos. La circulación del SARS-CoV-2 se evaluó por RT-qPCR y RT-PCR en muestras de frotis nasofaríngeos y orofaríngeos de gatos y perros cuyos propietarios se encontraban dentro del periodo de los 14 días de aislamiento. Los casos positivos se verificaron amplificando fragmentos de los genes RdRp, N y E; se secuenció el gen RdRp y se analizó filogenéticamente. Resultados. De 80 animales evaluados, seis gatos y tres perros fueron casos confirmados de infección natural por SARS-CoV-2. Los animales no presentaron signos clínicos y sus propietarios, que padecían la infección, reportaron únicamente signos leves de la enfermedad sin complicaciones clínicas. En el análisis de una de las secuencias, se encontró un polimorfismo de un solo nucleótido (SNP) con un cambio en la posición 647, con sustitución del aminoácido serina (S) por una isoleucina (I). Los casos se presentaron en los municipios de Caldas, Medellín y Envigado. Conclusiones. Se infiere que la infección natural en los gatos y perros se asocia al contacto directo con un paciente con COVID-19. No obstante, no es posible determinar la virulencia del virus en este huésped, ni su capacidad de transmisión zoonótica o entre especie.

Introduction: The severe acute respiratory syndrome caused by the new coronavirus SARS-CoV-2 is the cause of the health emergency due to the COVID-19 pandemic. Although humans are the main susceptible host, experimental studies and reported cases of natural infection have evidenced scenarios of SARS-CoV-2 reverse zoonosis in animals. Objective: To evaluate the natural infection of SARS-CoV-2 in cats and dogs with owners diagnosed with COVID-19 in the Valle de Aburrá subregion in Antioquia, Colombia. Materials and methods. The circulation of SARS-CoV-2 was evaluated by RT-qPCR and RT-PCR in samples of nasopharyngeal and oropharyngeal smears from cats and dogs whose owners presented latent COVID-19 infection. Positive cases were verified through amplification of N, E and RdRp gene fragments; with the latter being sequenced and the phylogenetically analyzed. Results. From 80 tested animals, 6 cats and 3 dogs resulted positive for natural SARS-CoV-2 infection. These animals did not show any clinical signs; and their infected owners only reported mild signs of COVID-19, without clinical complications. Regarding analysis of one of the sequences, a single nucleotide polymorphism (SNP) was found, with a substitution in position 647, resulting in the change of the amino acid serine (S) for isoleucine (I). The cases occurred in the municipalities of Caldas, Medellín and Envigado. Conclusions. It is inferred that natural infection in cats and dogs is associated with direct contact with a positive COVID-19 patient.

Virulence factors and phylotyping of Escherichia coli isolated from non-diarrheic and diarrheic water buffalo calves / Caracterização dos fatores de virulência e filogrupos de Escherichia coli isoladas de bezerros bubalinos sem diarreia e diarréicos

ABSTRACT: This study aimed to determine the virulence factors, phylogenetic groups, and the relationships between pathovars and phylogenetic groups of E. coli strains isolated from feces of buffalo calves. A total of 217 E. coli strains were obtained from feces after culture and were screened by PCR for detection of virulence factors EAST-1, enterohemolysin, Saa, CNF2, F41, F5, STa, intimin, Stx1 and Stx2. One hundred and thirty-four isolates were positive for one or more virulence factors: eighty-four from diarrheic animals, and fifty from non-diarrheic calves. The pathovars of E. coli identified in diarrheic feces were ETEC (F5+) (2/84), NTEC (16/84), STEC (20/84), EPEC (3/84), EHEC (3/84), and EAEC (EAST-1+) (33/84). Pathovars identified in non-diarrheic animals were NTEC (21/50), STEC (17/50), EHEC (1/50) and EAEC (7/50). E. coli strains positive for EAST-1 (P=0.008) and phylogroup C (P = 0.05) were associated with the presence of diarrhea. Phylogenetic analysis showed that 58.95% of the isolates belonged to phylogroup B1, followed by E (9.70%), B2 (5.90%), C (5.90%), D (5.22%), A (2.24%), and F (1.50%). Phylogroup B1 predominated in pathogenic E. coli isolated from water buffalo, and phylogroup C constituted an enteropathogenic E. coli for water buffalo calves.

RESUMO: O objetivo foi determinar os fatores de virulência, os grupos filogenéticos e as possíveis relações entre os patovares e os grupos filogenéticos identificados de cepas de Escherichia coli isoladas de fezes de bezerros bubalinos. Um total de 217 amostras de E. coli foram identificadas a partir de cultura das fezes e submetidas a reação em cadeia da polimerase (PCR) para detecção dos fatores de virulência EAST-1, enterohemolisina, Saa, CNF2, F41, F5, STa, intimina, Stx1 e Stx2. Foram identificadas 134 cepas positivas para um ou mais fatores de virulência: 84isoladas de bezerros bubalinos diarreicos e 50 de bezerros bubalinos saudáveis. Os patovares de E. coli obtidos de fezes diarreicas foram ETEC (F5+) (2/84), NTEC (16/84), STEC (20/84), EPEC (3/84), EHEC (3/84), e EAEC (EAST-1+) (33/84). Os patovares isolados de fezes não diarreicas foram NTEC (21/50), STEC (17/50), EHEC (1/50) e EAEC (7/50). Cepas de E. coli positivas para EAST-1 (P = 0,008) e filogrupo C (P = 0,05) foram associadas com a presença de diarreia. A análise de filogrupos revelou que 58,95% dos isolados pertencem ao filogrupo B1, seguido por E (9,70%), B2 (5,90%), C (5,90%), D (5,22%), A (2,24%) e F (1,50%). O filogrupo B1 predomina em cepas de E. coli patogênicas isoladas de bezerros búfalos e o filogrupo C constitui um filogrupo de E. coli patogênica entérica para bezerros.

Phylogenetic structure is determined by patch size in rock outcrop vegetation on an inselberg in the northern Amazon region / Estrutura filogenética é determinada pelo tamanho de manchas de vegetação sobre afloramento rochoso em um inselberg na região norte da Amazônia

Although inselbergs from around the world are iconic ecosystems, little is known on the underlying mechanisms of community assembly, especially in their characteristic patchy outcrop vegetation. Environmental constraints are expected to cause phylogenetic clustering when ecological niches are conserved within evolutionary lineages. We tested whether vegetation patches from rock outcrops of the Piedra La Tortuga Natural Monument, in the northern Amazon region, are phylogenetically clustered, indicating that environmental filtering is the dominant driver of community assemblage therein. We classified all patches according to their size as very small (< 1 m2), small (1-4 m2), medium-sized (4-8 m2), and large patches (8-15 m2). From each class, we randomly selected 10 patches, totalizing 40 patches covering 226 m2. All individuals found in the 40 isolated patches were identified to the species level. We also correlated measurements of phylogenetic community structure with patch size. We found that species from patches are restricted to the clades monocots, fabids, malvids, and lamiids. We conclude that vegetation in this rock outcrop is phylogenetically clustered. Furthermore, we found that phylogenetic turnover between pairs of patches increases with patch size, which is consistent with a scenario of higher environmental stress in smaller patches. Further research is necessary to identify nurse species in inselberg vegetation, which is pivotal for conservation and restoration of this particular ecosystem.

Ainda que os inselbergs ao redor do mundo sejam ecossistemas icônicos, pouco se sabe sobre os mecanismos subjacentes que estruturam suas comunidades vegetais, especialmente nas manchas de vegetação sobre afloramentos rochosos. Espera-se que as restrições ambientais causem agrupamento filogenético quando os nichos ecológicos são conservados dentro das linhagens evolutivas. Nós testamos se as manchas de vegetação dos afloramentos rochosos do Monumento Natural Piedra La Tortuga, no norte da região amazônica, apresentam indicadores filogenéticos de que a filtragem ambiental é o principal direcionador da estruturação da comunidade. Classificamos todas as manchas de acordo com seu tamanho como muito pequenas (<1 m2), pequenas (1-4 m2), médias (4-8 m2) e grandes (8-15 m2). Selecionamos aleatoriamente 10 manchas em cada classe de tamanho, totalizando 40 manchas cobrindo 226 m2. Todos os indivíduos encontrados nas 40 manchas foram identificados ao nível de espécie. Correlacionamos as medidas da estrutura filogenética da comunidade com o tamanho das manchas e encontramos que as espécies das manchas são restritas aos clados das monocotiledôneas, fabídeas, malvídeas e lamiídeas. Concluímos que a vegetação neste afloramento rochoso é agrupada filogeneticamente. Além disso, encontramos que o turnover filogenético entre pares de manchas aumenta com o tamanho da mancha, o que é consistente com um cenário de alto estresse ambiental nas manchas menores. São necessárias mais pesquisas para identificar espécies facilitadoras, que são fundamentais para a conservação e restauração destes ecossistemas.

Molecular epidemiology and phylogenetic analysis of human papillomavirus infection in women with cervical lesions and cancer from the coastal region of Ecuador / Epidemiología molecular y análisis filogenético de la infección por el virus del papiloma humano en mujeres con lesiones cervicales y cáncer en la región litoral del Ecuador

The aim of the present study was to gather information regarding the molecular epidemiology of Human papillomavirus (HPV) and related risk factors in a group of women with low- and high-grade cervical lesions and cancer from the coastal region of Ecuador. In addition, we studied the evolution of HPV variants from the most prevalent types and provided a temporal framework for their emergence, which may help to trace the source of dissemination within the region. We analyzed 166 samples, including 57 CIN1, 95 CIN2/3 and 14 cancer cases. HPV detection and typing was done by PCR-sequencing (MY09/MY11). HPV variants and estimation of the time to most recent common ancestor (tMRCA) was assessed through phylogeny and coalescence analysis. HPV DNA was found in 54.4% of CIN1, 74.7% of CIN2/3 and 78.6% of cancer samples. HPV16 (38.9%) and HPV58 (19.5%) were the most prevalent types. Risk factors for the development of cervical lesions/cancer were the following: three or more pregnancies (OR = 4.3), HPV infection (OR = 3.7 for high-risk types; OR = 3.5 for HPV16), among others. With regard to HPV evolution, HPV16 isolates belonged to lineages A (69%) and D (31%) whereas HPV58 isolates belonged only to lineage A. The period of emergence of HPV16 was in association with human populations (tMRCA = 91 052 years for HPV16A and 27000 years for HPV16D), whereas HPV58A preceded Homo sapiens evolution (322 257 years). This study provides novel data on HPV epidemiology and evolution in Ecuador, which will be fundamental in the vaccine era.

El objetivo del presente estudio fue aportar información sobre la epidemiología molecular del virus del papiloma humano (human papillomavirus [HPV]) y los factores de riesgo asociados al desarrollo de lesiones cervicales y cáncer en mujeres de la costa del Ecuador. Además, se estudiaron la evolución de las variantes de los HPV más prevalentes y el marco temporal de su emergencia, para ayudar a rastrear la fuente de dispersión en la región. Se analizaron 166 muestras, incluyendo 57 y 95 casos de neoplasia intraepitelial cervical tipo 1 (CIN1) y tipo 2/3 (CIN2/3), respectivamente, y 14 de casos de cáncer. La detección/tipificación de HPV se realizó por PCR-secuenciación (MY09/MY11). La caracterización de variantes y la datación del ancestro común más reciente (tMRCA) se realizaron mediante filogenia y coalescencia. Se encontró ADN de HPV en el 54,4% de las muestras de CIN1, el 74,7% de las muestras de CIN2/3 y el 78,6% de las muestras de cáncer. Los tipos HPV16 (38,9%) y HPV58 (19,5%) fueron los más frecuentes. Los factores de riesgo para el desarrollo de lesiones cervicales/cáncer fueron 3 o más embarazos (OR = 4,3) e infección por HPV (O = 3,7 para HPV de alto riesgo, OR = 3,5 para HPV16), entre otros. En cuanto a la evolución viral, los aislados del HPV16 pertenecían a los linajes A (69%) y D (31%), mientras que los aislados del HPV58 pertenecían únicamente al linaje A. El período de emergencia del HPV16 estuvo asociado a poblaciones humanas (tMRCA = 91.052 años para HPV16Ay 27.000 para HPV16D), mientras que el del HPV58A precedió a la evolución de Homo sapiens (322.257 años). Este estudio proporciona datos novedosos sobre la epidemiología y la evolución del HPV en Ecuador, los cuales serán fundamentales en la era de la vacuna.

Presence of bovine leukemia virus genotypes 1 and 3 in antioquia, colombia / Presencia de los genotipos 1 y 3 del virus de la leucosis bovina en antioquia, colombia

SUMMARY Bovine leukemia virus (BLV) is an immunosuppressant retrovirus that primarily affects dairy livestock, its target cells are B lymphocytes in which it integrates its genome infecting cattle for life. It is important to identify the distribution of the BLV in the region and to reconstruct its evolutionary history through phylogenetic trees, for the province of Antioquia this is the first report of the BLV genotypes. The aim of this study was to identify the genotype of BLV circulating in dairy cattle of different regions of the province of Antioquia, Colombia. DNA was extracted from 8 Holstein cows. Nested PCR was performed to amplify a fragment of 444 pb of the env gene. The env viral gene codes for surface protein gp51, gene is highly conserved and it is used for phylogenetic analysis. Obtained amplicons were sequenced, manually aligned in MEGA V7 program, and compared to 53 viral env gene sequences registered in GenBank. Phylogenetic analysis was performed by Maximum Likelihood and Bayesian methods. Two circulating genotypes were found: the most common genotype was 1, found in seven samples; they grouped with sequences from EE. UU, Argentina and Japan; only one sample was classified as genotype 3 and was grouped with samples from EE. UU and Japan. At least two genotypes (1 and 3) of BLV are circulating in Antioquia; however, more cattle and herds should be evaluated to elucidate the diversity and distribution of BLV in Colombia.

RESUMEN El virus de la leucosis bovina (BLV) es un retrovirus inmuno-supresor que afecta, principalmente, al ganado lechero; sus células diana son los linfocitos B, en los cuales, integra su genoma, infectando al ganado de por vida. Es importante identificar la distribución del BLV en la región y reconstruir su historia evolutiva, a través de árboles filogenéticos; para el departamento de Antioquia, este es el primer reporte de los genotipos del BLV. El objetivo de este estudio fue identificar el genotipo de BLV que circula en ganado lechero de diferentes regiones del departamento de Antioquia, Colombia. Se extrajo ADN de 8 vacas Holstein. Se realizó una PCR anidada, para amplificar un fragmento del gen env de 444 pb. El gen env viral codifica la proteína de superficie gp51, altamente conservado y es usado en análisis filogenéticos. Los amplicones obtenidos se secuenciaron, se alinearon manualmente en el programa MEGA V7 y se compararon con 53 secuencias del gen env viral, registradas en GenBank. El análisis filogenético, se realizó por métodos de Máxima Verosimilitud y Bayesianos. Se encontraron dos genotipos circulantes: el genotipo más común fue 1, hallado en siete muestras, agrupadas con secuencias de EE. UU, Argentina y Japón; solo una muestra se clasificó como genotipo 3 y se agrupó con muestras de EE. UU y Japón. Al menos dos genotipos (1 y 3) de BLV están circulando en Antioquia; sin embargo, se deben evaluar más bovinos y hatos para elucidar la diversidad y la distribución de BLV en Colombia.

Implicaciones de la precipitación sobre la evolución del tamaño corporal y distancia interaxilar en el complejo Aspidoscelis gularis (Squamata: Teiidae) / Rainfall implications on body size evolution of Aspidoscelis gularis (Squamata: Teiidae)

ResumenLas historias de vida son atributos altamente variables que maximizan la adecuación de los organismos. Se ha documentado la relación del peso y el tamaño corporal sobre estos atributos, los cuales además estan influidos por variaciones ambientales y la heterogeneidad del hábitat. El tamaño y forma corporal son considerados atributos de historia de vida, los cuales estan principalmente asociados a la precipitación que promueve la cantidad de recursos disponibles en el ambiente. En el género Aspidoscelis el tamaño y masa relativa de la nidada estan asociados principalmente a la latitud y altitud. En Aspidoscelis gularis la precipitación de invierno favorece dos temporadas reproductoras, lo que determina que el tamaño de nidada sea variable en cada puesta. Las muestras fueron tomadas de mayo-julio 2013 y mayo-septiembre 2015. Se obtuvieron un total de 65 individuos de los cuales se tomaron medidas de longitud del cuerpo y distancia interaxilar, así como muestras de tejido hepático para la extracción de ADN y obtener las relaciones filogenéticas con respecto a un análisis de Inferencia Bayesiana para posteriormente aplicar los Métodos Comparados Filogenéticos como señal filogenética, contrastes filogenéticamente independientes y la reconstrucción del carácter ancestral. Por lo tanto, nuestros resultados muestran que en el tamaño y forma del cuerpo existe poca señal filogenética, en tanto que los contrastes filogenéticamente independientes y la reconstrucción del carácter ancestral sugieren que los tamaños corporales pequeños están asociadas a localidades con mayor cantidad de precipitación, lo que puede estar relacionado con un establecimiento de la madurez sexual temprana lo cual es el reflejo de la talla máxima de los adultos. Además, de acuerdo a un ANOVA y un ANCOVA hubo diferencias estadísticamente significativas en el tamaño y forma corporal respectivamente, lo que promueve un sistema para la competencia sexual para machos y un sistema para la fecundidad en hembras. Estos resultados son importantes para determinar el efecto de la precipitación sobre algunas características de historia de vida, lo que indica que las lagartijas del complejo sureste del complejo A. gularis son capaces de enfrentar diferentes presiones de selección, impuestas por el ambiente.

AbstractLife history traits are highly variable attributes that maximize organisms's fitness. The relationship of weight and body size with environmental changes and habitat heterogeneity has been documented in previous reports; and size and body shapes are both considered life history attributes that are associated with rainfall, that boost available resources in the environment. While in Aspidoscelis genus, clutch size and relative mass are mainly associated with latitude and altitude, in Aspidoscelis gularis, winter rainfall favors two reproductive seasons, which may determine season variable clutch size. With the aim to study this, samplings were undertaken from May-July 2013, and May-September 2015. A total of 65 individuals lizards of the Southeast clade were obtained, and body length and interaxilar distance measurements were taken; furthermore, hepatic tissue samples were taken for DNA extraction, which allowed us to analyze phylogenetic relationships through a Bayesian Inference analysis, and subsequently, to apply Phylogenetic Comparative Methods (like phylogenetic signal, phylogenetically independent contrasts and reconstruction of ancestral character). Our results showed that there is a low phylogenetic signal regarding body size and shape, while the phylogenetically independent contrasts and reconstruction of ancestral characters suggest that small body sizes are associated to locations with highest rainfall. This can be associated to an establishment of an early sexual maturity, which reflects the maximum size of adults. Furthermore, according to an ANOVA and ANCOVA, there were statistically significant differences in body size and shape respectively, which promote a system for sexual competition for males and a system for fertility in females. These results were important to determine the effect of rainfall on some life history traits, pointing out that lizards of the Southeast clade, belonging to the A. gularis complex were able to face different selection pressures, determined by the environment.

Phylogeny of Dengue virus type 2 isolated in the Central Highlands, Vietnam / Filogenia del virus del Dengue tipo2 identificado en el Altiplano Central de Vietnam

AbstractDengue fever is perhaps the most important viral re-emergent disease especially in tropical and sub-tropical countries, affecting about 50 million people around the world every year. In the Central Highlands regions of Vietnam, dengue fever still remains as a major public health issue. Although four viral serotypes have been currently identified, dengue virus type 2 (DENV-2) was involved in the most important outbreaks during 2010-2012, especially, 2010 when the fatality rate highly increased. Detection of genotype of DENV2 provided information on origin, distribution and genotype of the virus. In this study, DEN-2 isolated from dengue patients during the 2010-2012 epidemics was amplified and sequenced with E gene. The consensus sequences were aligned with reference E gene sequences of globally available Genbank. Phylogenetic analysis was performed using Neighbor-joining and Kimura 2-parameter model to construct phylogenetic tree. A total of 15 isolates (seven from 2010; one from 2011 and seven from 2012) were obtained from human serum samples. Phylogenetic analysis revealed that Asian genotype 1 is currently circulating locally in Central Highlands region. Isolates of this genotype were closely related to viruses from Thailand, Laos, and Cambodia. It indicated that these epidemics maybe imported into the Central Highlands region from South-East Asia neighbor countries. The study results would help in planning for prevention and control of dengue virus in Vietnam. Continuous monitoring of DENV genotypes is necessary to confirm the current findings and detect possible genotype shifts within the dengue viruses in the future.

ResumenPosiblemente, la fiebre del dengue es la enfermedad viral recurrente más importante en los países tropicales y subtropicales que afecta cerca de 50 millones de personas cada año en todo el mundo. En las regiones del Altiplano Central de Vietman, la fiebre del dengue aun se considera como una gran preocupación de salud pública. Aunque los cuatro serotipos virales han sido identificados, el virus del dengue tipo 2 (DENV-2) estuvo involucrado en el brote más importante durante el 2010-2012, especialmente en el 2010 cuando los índices de mortalidad aumentaron considerablemente. El descubrimiento del genotipo DENV-2 proporcionó información del origen, distribución y genotipo del virus. En este estudio, el serotipo DENV-2 identificado de pacientes con dengue durante las epidemias de 2010-2012 se amplificaron y secuenciaron con E gene. Las secuencias consenso se alinearon con secuencias de referencia mundiales de E gene disponibles en GenBank. El análisis filogenético se llevó a cabo utilizando el modelo Neighbor-joining y Kimura 2-parámetros para construir el árbol filogenético. Un total de 15 cepas (siete de 2010, una de 2011 y 7 de 2012) se obtuvieron de las muestras de suero humano. El análisis filogenético reveló que el genotipo asiático 1 circula localmente en la región del Altiplano Central. Las cepas de este genotipo estan muy relacionadas con los virus de Tailandia, Laos y Camboya. El análisis también indicó que estas epidemias pudieron migrar a la región del Altiplano Central desde los países vecinos del sureste asiático. Los resultados de este estudio pueden ayudar en la planificación de la prevención y el control del virus del dengue en Vietnam. Un monitoreo contante de los genotipos DENV es necesario para confirmar los hallazgos recientes y detectar los posibles cambios del genotipo de los virus del dengue en el futuro.

Detection of virulence factors and antimicrobial resistance patterns in shiga toxin-producing Escherichia coli isolates from sheep / Detecção de fatores de virulência e de resistência antimicrobiana em isolados produtores de toxina Shiga de ovinos

In order to detect virulence factors in Shiga toxin-producing Escherichia coli (STEC) isolates and investigate the antimicrobial resistance profile, rectal swabs were collected from healthy sheep of the races Santa Inês and Dorper. Of the 115 E. coli isolates obtained, 78.3% (90/115) were characterized as STEC, of which 52.2% (47/90) carried stx1 gene, 33.3% (30/90) stx2 and 14.5% (13/90) both genes. In search of virulence factors, 47.7% and 32.2% of the isolates carried the genes saa and cnf1. According to the analysis of the antimicrobial resistance profile, 83.3% (75/90) were resistant to at least one of the antibiotics tested. In phylogenetic classification grouped 24.4% (22/90) in group D (pathogenic), 32.2% (29/90) in group B1 (commensal) and 43.3% (39/90) in group A (commensal). The presence of several virulence factors as well as the high number of multiresistant isolates found in this study support the statement that sheep are potential carriers of pathogens threatening public health...

A fim de detectar os fatores de virulência em isolados de E. coli produtoras de toxina Shiga (STEC) e investigar o perfil de resistência aos antimicrobianos, swabs retais foram coletados em ovelhas saudáveis das raças Santa Inês e Dorper. Dos 115 isolados de E. coli obtidos, 78,3% (90/115) foram caracterizados como STEC, dos quais 52,2% (47/90) possuíam o gene stx1, 33,3% (30/90) stx2 e 14,5% (13/90) ambos os genes. Em busca de fatores de virulência, 47,7% e 32,2% dos isolados apresentaram genes saa e cnf1. De acordo com a análise do perfil de resistência a antimicrobianos, 83,3% (75/90) eram resistentes a pelo menos um dos antibióticos testados. Na classificação filogenética, os isolados foram agrupados 24,4% (22/90) no grupo D (patogênico), 32,2% (29/90) no grupo B1 (comensal) e 43,3% (39/90) no grupo A (comensal). A presença de vários fatores de virulência, bem como o elevado número de isolados multirresistentes encontrados neste estudo apoia a afirmação de que as ovelhas são portadoras potenciais de patógenos que ameaçam a saúde pública...

Variaciones nucleotídicas de dos grupos de tepezcuintles, Agouti paca (Rodentia: Agoutidae), en cautiverio provenientes de dos localidades de Yucatán, México / Nucleotidic variations of two captive groups of tepezcuintle, Agouti paca (Rodentia: Agoutidae), from two sites in Yucatan, Mexico

The objective of this work was to estimate the nucleotidic variation between two groups of tepezcuintles (Agouti paca) from the states of Campeche and Quintana Roo, Mexico and within members of each group. Blood samples were collected from eleven A. paca kept in captivity. DNA from leukocytic cells was used for Ramdom Amplification of DNA Polimorphism (RAPD). The primers three 5'-d(GTAGACCCGT)- 3' and six 5'-d(CCCGTCAGCA)- 3' were selected from de Amersham kit (Ready.To.Go. RAPD Analysis Beads, Amersham Pharmacia Biotech), because they produced an adequate number of bands. The electrophoretic pattern of bands obtained was analyzed using software for phylogenetic analysis based on the UPGMA method, to estimate the units of nucleotidic variation. The phylogenetic tree obtained with primer three reveals a dicotomic grouping between the animals from both states in the Yucatan Peninsula showing a divergent value of 1.983 nucleotides per hundred. Animals from Quintana Roo show a grouping with primer six; an additional grouping was observed with animals from Campeche. Nucleotidic variation between both groups was 2.118 nucleotides per hundred. The nucleotidic variation for the two primers within the groups from both states, showed fluctuating values from 0.46 to 1.68 nucleotides per hundred, which indicates that nucleotidic variation between the two groups of animals is around two nucleotides per hundred and, within the groups, less than 1.7 nucleotides per hundred.

Estimamos las variaciones nucleotídicas entre dos grupos de tepezcuintles (Agouti paca) provenientes de los estados de Campeche y Quintana Roo, México y, dentro de cada grupo. Se colectaron muestras sanguíneas de once A. paca mantenidos en cautiverio. El ADN de leucocitos se utilizó para efectuar la amplificación aleatoria de polimorfismos de ADN (RAPD). Se seleccionaron los iniciadores número tres 5’ -d(GTAGACCCGT)-3’ y seis 5’ -d(CCCGTCAGCA)-3’ del estuche (Ready.To.Go. RAPD Analysis Beads, Amersham Pharmacia Biotech), porque produjeron un adecuado número de bandas. Los patrones electroforéticos de bandas fueron procesados con el software para análisis filogenético basado en el método de UPGMA para estimar la variación nucleotídica. El árbol filogenético obtenido con el iniciador tres reveló una agrupación dicotómica entre los animales de ambos estados de la Península de Yucatán, con un valor de divergencia de 1.983 nucleótidos de cada cien. Los animales de Quintana Roo mostraron un agrupamiento con el iniciador seis y, otro grupo más con animales procedentes de Campeche. La variación nucleotídica entre estos dos grupos fue de 2.118 nucleótidos por cada cien. Las variaciones nucleotídicas dentro de los grupos procedentes de ambos estados, para los dos iniciadores, mostraron valores que fluctuaron entre 0.46 y 1.68 nucleótidos de cada cien, lo cual indica que la variación nucleotídica entre los dos grupos de animales es alrededor de dos nucleótidos por cada cien y, dentro de grupos es menor a 1.7 nucleótidos por cada cien.

Phylogenetic analysis of the envelope protein (domain lll) of dengue 4 viruses

Objective. To evaluate the genetic variability of domain III of envelope (E) protein and to estimate phylogenetic relationships of dengue 4 (Den-4) viruses isolated in Mexico and from other endemic areas of the world. Material and Methods. A phylogenetic study of domain III of envelope (E) protein of Den-4 viruses was conducted in 1998 using virus strains from Mexico and other parts of the world, isolated in different years. Specific primers were used to amplify by RT-PCR the domain III and to obtain nucleotide sequence. Based on nucleotide and deduced aminoacid sequence, genetic variability was estimated and a phylogenetic tree was generated. To make an easy genetic analysis of domain III region, a Restriction Fragment Length Polymorphism (RFLP) assay was performed, using six restriction enzymes. Results. Study results demonstrate that nucleotide and aminoacid sequence analysis of domain III are similar to those reported from the complete E protein gene. Based on the RFLP analysis of domain III using the restriction enzymes Nla III, Dde I and Cfo I, Den-4 viruses included in this study were clustered into genotypes 1 and 2 previously reported. Conclusions. Study results suggest that domain III may be used as a genetic marker for phylogenetic and molecular epidemiology studies of dengue viruses.

Objetivo. Evaluar la variabilidad genética del dominio III de la proteína de envoltura (E) y estimar la relación filogenética de los virus dengue 4 (Den-4) aislados en México y en otras regiones endémicas del mundo. Material y métodos. En el presente trabajo reportamos un estudio filogenético del dominio III de la proteína de envoltura (E) que se realizó en 1998 con virus Den-4 aislados en distintos años en México y en otras partes del mundo. Se usaron oligonucleótidos específicos para amplificar por RT-PCR la región del dominio III y para obtener la secuencia de nucleótidos. Mediante el análisis de la secuencia de nucleótidos y de la secuencia deducida de aminoácidos se estimó la variabilidad genética y se generó un árbol filogenético. Para facilitar el análisis genético del dominio III se usó la técnica basada en el polimorfismo de fragmentos generados con enzimas de restricción (PFER) utilizando seis enzimas de restricción. Resultados. Los datos demuestran que la información del análisis de la secuencia de nucleótidos y de aminoácidos de la región del dominio III es similar a la del gene completo de la proteína E. El análisis de PFER con las enzimas de restricción Nla III, Dde I y Cfo I, mostró que los virus Den-4 incluidos en este estudio se agruparon en los genotipos 1 y 2 reportados previamente. Conclusiones. Los resultados sugieren que el dominio III se puede utilizar como un marcador para estudios filogenéticos y de epidemiología molecular del virus Den-4.