Detection of the ompA gene of Chlamydophila pecorum in captive birds in Argentina / Detección del gen ompA de Chlamydophila pecorum en aves en cautiverio, en la Argentina

Bacteria belonging to the family Chlamydiaceae cause a broad spectrum of diseases in a wide range of hosts, Including humans, other mammals and birds. However, very little is known about chlamydial infections in birds in our region. In the present study, we examined 28 clinically normal birds In illegal captivity that were confiscated in the province of Córdoba, Argentina. The objective was to detect Chlamydophila spp. in cloacal swabs by genetic analysis of the ompA gene. Nested-PCR of the ompA gene identified five samples as Chlamydophila pecorum and the sequence analysis demonstrated the presence of the ompA gene of C. pecorum In these birds. On the other hand, Chlamydophila psittaci was not detected. These birds could be either asymptomatic reservoirs or subclinical carriers of C. pecorum. This is the first report of the detection of C. pecorum in Argentina.

Las bacterias que pertenecen a la familia Chlamydiaceae causan un extenso espectro de enfermedades en una amplia gama de huéspedes, incluidos los seres humanos, otros mamíferos y aves. Sin embargo, se sabe muy poco acerca de las infecciones por clamidias en aves de nuestra reglón. Esta Investigación examinó 28 aves clínicamente normales mantenidas en cautiverio ¡legal, que fueron confiscadas en Córdoba, Argentina. El objetivo fue detectar Chlamydophila spp. en hisopados cloacales por análisis del gen ompA. La PCR anidada del gen ompA reveló la presencia de Chlamydophila pecorum en cinco muestras. El análisis de secuencias demostró la presencia del gen ompA de C. pecorum en estas aves. Por el contrario, Chlamydophila psittaci no se detectó. Estas aves pueden ser reservónos asintomáticos o portadores subclínlcos de C. pecorum. Este es el primer informe de la detección de C. pecorum en la Argentina.

Evidencia morfológica de hibridación entre las subespecies de Ramphocelus flammigerus (Passeriformes: Thraupidae) en Colombia / Morphological evidence of hybridization between Ramphocelus flammigerus subspecies (Passeriformes: Thraupidae) in Colombia

Morphological evidence of hybridization between Ramphocelus flammigerus subspecies (Passeriformes: Thraupidae) in Colombia. Habitat modifications such as deforestation and the increase of agricultural activities, have led to uncommon faunal interactions. In Colombia, this condition have caused the secondary contact of subspecies of Ramphocelus flammigerus populations from Cauca valley and the Pacific coast; and some specimens with rumps of intermediate colors of the subspecies have been found and are thought as hybrids. The objective of this study was to assess the presence of morphological evidence that may suggest hybridization and may explain the origin of individuals with intermediate coloration. We predict that if subspecies hybridize, they will be more similar in morphology when coexisting than when separated. Alternatively, coexisting subspecies might diverge in sympatry, because of selection to reduce competition for resources (character displacement). For this, a survey in 15 localities was undertaken: 10 allopatric areas (five for each subspecies), and five sympatric areas. Mist nets were used to capture individuals and a total of seven morphological characters were measured. To identify the patterns of morphological variation, we compared morphology of subspecies, sympatric and allopatric populations and individuals of intermediate colors. Consequently, we performed discriminant analysis and test for differences between groups by using 95% confidence intervals for log-ratio tests. A total of 112 individuals were captured (46 intermediate-colored individuals, 20 R. f. flammigerus, and 46 R. f. icteronotus. Discriminant analyses showed that subspecies were well differentiated, and intermediate individuals overlapped with them. Log-ratio test, based on Mahalanobis distances, showed that intermediate individuals were morphologically more similar to both subspecies than subspecies themselves. In addition, log-ratio tests showed that subspecies sympatric populations were similar but allopatric ones were different, and that individuals of intermediate colors were more similar to sympatric than to allopatric populations of the two subspecies. Therefore, morphological evidence supports the predictions of a hybridization hypothesis among the subspecies of R. flammigerus. In conclusion, the analysis of morphological variation in R. flammigerus suggests that hybridization between subspecies is occurring and that a process of genetic introgression is probably in progress.

Las modificaciones de los hábitats naturales, tales como la deforestación y el incremento de las actividades agrícolas, han conducido a interacciones faunísticas inusuales. En Colombia, esta situación ha generado el contacto secundario entre las poblaciones de las subespecies de Ramphocelus flammigerus del Valle del Cauca y de la costa Pacífica; y se han encontrado individuos con rabadillas de colores intermedios entre las subespecies que se han catalogado como híbridos. El objetivo del presente estudio fue evaluar si existe evidencia morfológica que sugiera hibridación y que pueda explicar el origen de los individuos de coloración intermedia. Con este fin, se obtuvieron muestras de 15 localidades; 10 zonas alopátricas (cinco por cada subespecie) y cinco zonas simpátricas. Para la captura de individuos se utilizaron redes de niebla y fueron tomados siete caracteres morfológicos. Asimismo, se predijo que si las subespecies están hibridando, las mismas, podrían ser morfológicamente más similares cuando coexisten que cuando se encuentran separadas. Alternativamente, cuando las subespecies coexisten, éstas pueden divergir en simpatría debido a presiones selectivas para reducir la competencia por recursos (desplazamiento de caracteres). Para identificar los patrones de variación morfológica, se comparó la morfología de las subespecies, de poblaciones simpátricas y alopátricas de ambas subespecies y de los individuos de cloración intermedia. Consecuentemente, se realizó un análisis discriminante y se evaluaron las diferencias entre los grupos con la utilización de intervalos de confianza del 95% para las relaciones logarítmicas. Y se capturaron un total de 112 individuos (46 de coloraciones intermedias, 20 R. f. flammigerus y 46 R. f. icteronotus). Los análisis discriminantes mostraron que las subespecies se diferencian entre ellas y que los individuos de coloraciones intermedias se traslapan con estas. Las relaciones logarítmicas, basadas en las distancias cuadradas de Mahalanobis, mostraron que los individuos intermedios fueron más similares morfológicamente a ambas subespecies que las subespecies entre ellas. Adicionalmente, se encontró que las poblaciones simpátricas de ambas subespecies son similares pero las poblaciones alopátricas son diferentes, y que los individuos de coloraciones intermedias son más similares a las poblaciones simpátricas de las dos subespecies que a las alopátricas. Por lo tanto, la evidencia morfológica evidencia las predicciones derivadas de la hipótesis de hibridación entre las subespecies de R. flammigerus. En conclusión, el análisis morfológico de la variación en R. flammigerus sugiere que se presenta hibridación entre las subespecies y que probablemente está en progreso un proceso de introgresión genética.

Sex determination in Turdus amaurochalinus (Passeriformes: Muscicapidae): morphometrical analysis supported by CHD gene

Sex determination is important for conservation and population studies, particularly for reproduction programs of threatened species and behavioural ecology. Turdus amaurochalinus, Creamy-bellied Thrush, only exhibits sexual dimorphism during the breeding season, when males are considered to show intense yellow bills, and females and immature males show dark brown bills. The objectives of this study were: 1) to determine the sex of individuals using genetic techniques, and 2) to test the hypothesis that sex dimorphism can be detected by morphometry. This study was carried out at Parque Nacional da Restinga de Jurubatiba, a preserved area located on the North coast of Rio de Janeiro State. The birds were captured using ornithological nets, singly marked with metal rings, weighed, measured and had blood samples collected before being released. The sex of 42 T. amaurochalinus individuals was determined using the CHD gene marker. A total of 20 males and 22 females were identified from June to August, with peak capture frequency in June. Turdus amaurochalinus females and males differed significantly in morphometrical measures. The most important traits to distinguish males from females were wing length (Student t-test=4.34, df=40, p=0.0001) and weight (Student t-test=2.08,df=40, p=0.044): females were heavier and had significantly shorter wing length than males. Females and males were correctly classified in 86% and 75% of cases, respectively, using Discriminant Analysis. The molecular analysis was the most secure method for sex determination in the studied species. Rev. Biol. Trop. 59 (2): 789- 794. Epub 2011 June 01.

La determinación del sexo es importante para la conservación y los estudios poblacionales. Turdus amaurochalinus no presenta aparente dimorfismo sexual. El objetivo de este estudio fue determinar el sexo a través de una técnica genética, mediante el uso del marcador del gen CHD y se puso a prueba la hipótesis de que el dimorfismo sexual puede ser detectado por morfometría. Este estudio se llevó a cabo en el Parque Nacional da Restinga de Jurubatiba, una zona protegida situada en la costa norte de Río de Janeiro. Las aves fueron capturadas con redes de niebla, los individuos se marcaron con anillos de metal, se pesaron, medieron y se les tomó una muestra de sangre antes de ser liberados. Un total de 20 machos y 22 hembras fueron identificados en el área de estudio desde junio hasta agosto, con la frecuencia máxima de captura en junio. La prueba de t-student fue usada para evaluar si hembras y machos se diferencian considerablemente en relación a medidas morfométricas. Los rasgos más importantes para distinguir machos de hembras fueron la longitud del ala y el peso: las hembras eran más pesadas y tenían longitud de ala considerablemente más corta que los machos. Hembras y machos fueron correctamente clasificados en un 86% y 75% de casos respectivamente, donde se usó un análisis discriminante. El análisis molecular es el método más seguro para la determinación sexual en la especie estudiada.

Iris colour as an indicator of age feature in female Brazilian tanagers (Passeriformes: Emberizidae) confirmed by a molecular sexing technique

The Brazilian tanager, Ramphocelus bresilius is an endemic species from Brazil that is sexually dimorphic in adult plumage. Young males are similar to adult and young females until their second year. Adults and young females are not distinguishable in plumage. We tested whether iris colour can be used to separate adult females from immature females. We used for the first time the molecular sexing technique based on CHD-genes to confirm the sex of the individuals classified as "female plumage with red iris", and to identify the sex of individuals classified as "female plumage and brown iris". The adult males were used as a positive control. DNA samples from 190 individuals were analysed. The sizes of the PCR products were identified as 350 base pairs (bp) for CHD-Z and 388 bp for CHD-W. We confirmed that adult females have a red iris and the young females a brown iris. We could also separate young males and females which present the same iris colour and plumage. Although there are indications that the iris colour can be used by birds to identify the adults in co-operative breeding species such as the Brazilian tanager, more behavioural data are required to understand the role of iris coloration in this species. Rev. Biol. Trop. 56 (4): 1629-1633. Epub 2008 December 12.

El ave Ramphocelus bresilius es una especie endémica de Brasil con dimorfismo sexual en el plumaje del adulto. Los machos jóvenes son similares a las hembras adultas y jóvenes hasta el segundo año de vida. Adultos y hembras jóvenes son indistinguibles por el plumaje. Evaluamos si el color del iris puede ser utilizado para distinguir hembras adultas de hembras inmaduras. Utilizamos por primera vez la técnica molecular de identificación de sexos basada en los genes CHD para confirmar el género de individuos clasificados como plumaje femenino con iris rojo, y para identificar el sexo de los individuos clasificados como plumaje femenino e iris marrón. Usamos machos adultos como control. Analizamos muestras de DNA de 190 individuos. Los tamaños de los productos del PCR fueron identificados como 350 pares de bases (pb) para CHD-Z y 388 pb para CHD-W. Pudimos confirmar que las hembras adultas presentan iris rojo y las hembras jóvenes iris marrón. También pudimos distinguir machos jóvenes de hembras, que presentan el mismo color de iris y plumaje.

Genetic variability of Conopophaga lineata (Conopophagidae) (Wied-Neuwied, 1831) in Atlantic Forest fragments

Forest fragmentation affects bird populations in many ways, modifying the composition of communities and favouring open country species. The Atlantic Forest is considered one of the most important biomes in the world, due to its great biodiversity, accelerated rates of deforestation, and high endemism. Despite these characteristics, few studies have evaluated the effects of forest fragmentation in the genetic structure of Atlantic forest bird populations. So, this study aims to verify the effects of forest fragmentation in the genetic population structure of Conopophaga lineata, through RAPD markers. To achieve this goal, 89 C. lineata individuals were captured in nine Atlantic Forest fragments in Minas Gerais State. The RAPD data indicate that forest fragmentation has not affected the genetic variation of C. lineata populations (Mann-Whitney U = 3.50; p = 0.11). Great part of the genetic variability of this species is found within populations and it was not observed a correlation between genetic and geographic distance (Mantel test t = 0.6250; p = 073). UPGMA analyses did not show defined clades and all branches showed low statistical support. The low population differentiation observed in this species can be due to a high gene flow among populations or a recent fragmentation. Thus, the current diversity status of C. lineata populations indicates that this species is not significantly affected by fragmentation. However, more genetic studies are essential to improve conservation strategies of Brazilian Atlantic Forest birds.

A fragmentação florestal afeta populações de aves de muitas maneiras, modificando a composição das comunidades e favorecendo espécies de áreas abertas. A Mata Atlântica é considerada um dos biomas mais importantes do mundo, devido à sua grande biodiversidade, aos altos níveis endemismo e de desmatamento. Apesar destas características, poucos estudos avaliaram os efeitos da fragmentação florestal na estrutura genética de populações de aves desse ecossistema. Dessa forma, este estudo visa verificar os efeitos da fragmentação florestal na variabilidade genética de Conopophaga lineata, através de marcadores RAPD. Assim, foram capturados 89 indivíduos de C. lineata em nove fragmentos florestais da Mata Atlântica do Estado de Minas Gerais. Os dados de RAPD indicam que a fragmentação florestal não afetou a variabilidade genética de Conopophaga lineata (Mann-Whitney U = 3,50; p = 0,11). Grande parte da variabilidade dessa espécie se encontra dentro das populações e não foi observada correlação entre a distância geográfica e a distância genética (Teste de Mantel t = 0,6250; p = 0.73). A análise UPGMA não mostrou clados definidos e todos os ramos apresentaram baixo suporte estatístico. A reduzida diferenciação populacional observada nessa espécie pode ser devida a intenso fluxo gênico entre as populações ou à recente fragmentação na região. Assim, a situação atual das populações de C. lineata indica que essa espécie não é significativamente afetada pela fragmentação. Entretanto, futuros estudos genéticos são essenciais para melhorar as estratégias de conservação da avifauna da Mata Atlântica do Brasil.

Population structuring of the endemic black-cheeked gnateater, Conopophaga melanops melanops (Vieillot, 1818) (Aves, Conopophagidae), in the Brazilian Atlantic Forest

Randomly amplified polymorphic DNA (RAPD) markers were used to analyze genetic differentiation among three populations of the endemic Black-cheeked Gnateater (Conopophaga melanops melanops) within a larger pristine reminiscent of the Brazilian Atlantic Forest. Analyses of molecular variance (AMOVA) (phiST = 0.13149, P < 0.0001) and the nonparametric test for homogeneity of the molecular variance (HOMOVA) (B = 0.32337; P = 0.0019) showed a statistically significant genetic divergence among the three Black-cheeked Gnateater populations in a continuous transect of 250 km. Some hypothetic explanations for these results are the sedentary nature of the species and the historical isolation of the populations in refuges during the Pleistocene. The present results suggest that the local populations were naturally differentiated along the entire original range before the recent process of massive deforestation.

Marcadores polimórficos de DNA amplificados ao acaso (RAPD) foram utilizados para analisar a diferenciação genética entre três populações do endêmico cuspidor-de-máscara preta (Conopophaga melanops melanops) de um amplo remanescente preservado da floresta Atlântica brasileira. Análises de Variância Molecular (AMOVA) (fiST = 0,13149, P < 0,0001) e o teste não paramétrico para a homogeneidade da variância molecular (HOMOVA) (B = 0,32337; P = 0,0019) comprovaram uma divergência genética significativa entre as três populações em um transecto contínuo de 250 km. Algumas explicações hipotéticas para estes resultados são a natureza sedentária da espécie e o isolamento histórico das populações em refúgios durante o Pleistoceno. Os presentes resultados sugerem que as populações locais foram diferenciadas naturalmente ao longo da distribuição original antes do recente processo de intenso desmatamento.

Mitochondrial DNA corroborates the species distinctiveness of the Planalto (Thamnophilus pelzelni Hellmayr, 1924) and the Sooretama (T. ambiguus Swainson, 1825) Slaty-antshrikes (Passeriformes: Thamnophilidae)

The Thamnophilus punctatus complex has been recently reviewed on the basis of morphological and vocal characters, and is divided in six different species. Two of the new species, although well defined on the basis of morphological differences, could not be unambiguously distinguished through their loudsongs. The Planalto Slaty-antshrike (Thamnophilus pelzelni) and the Sooretama Slaty-antshrike (T. ambiguus) are most easily distinguished by subtle and localized changes in plumage colors of males and females. In the present study we used sequences of the control region, Cytochrome b, and ND2 genes, of the mitochondrial DNA (mtDNA) to evaluate the levels of molecular differentiation between these two species. The mean pairwise distance between the two species was 3.8 percent, while it varied from 2.7 percent to 4.9 percent for each mtDNA region. Although extensive variation was also detected among haplotypes within species, especially for T. ambiguus, we suggest that the genetic divergence found between T. ambiguus and T. pelzelni is high enough to corroborate the separate species status of these two antbird taxa.

O complexo Thamnophilus punctatus foi recentemente revisado, com base em caracteres morfológicos e vocais, sendo dividido em seis diferentes espécies. Duas dessas novas espécies, embora bem definidas com base em distinções morfológicas, não puderam ser definitivamente diferenciadas por seus cantos. Thamnophilus pelzelni (choca-do-planalto) e T. ambiguus (choca-de-sooretama) são mais facilmente diferenciadas por sutis e localizadas mudanças de coloração da plumagem de machos e fêmeas. Neste estudo, foram utilizadas seqüências de DNA mitocondrial (região controle, Citocromo b e ND2) a fim de avaliar os níveis de diferenciação molecular entre estas duas espécies. A divergência genética média entre as duas espécies foi de 3,8 por cento, enquanto para cada região mitocondrial esta variou entre 2,7 por cento e 4,9 por cento. Embora tenha sido observada grande variabilidade entre haplótipos dentro das espécies, especialmente para T. ambiguus, os resultados sugerem que a divergência genética observada entre T. ambiguus e T. pelzelni é suficientemente elevada para corroborar o status de espécies separadas destes dois Thamnophilidae.