RESUMEN
A total of 10 specimens were captured from selected sites of Bajaur Agency FATA, Pakistan using mist nets. The captured specimens were morphologically identified and various morphometric measurements were taken. The head and Body length (HB) of Pipistrellus coromondra and Pipistrellus kuhlii lepidus (n=10) was 43±0.11 mm and 45±1.1 respectively. Morphologically identified Pipistrellus kuhlii confirmed as Pipistrellus kuhlii lepidus based on 16S rRNA sequences. The DNA sequences were submitted to GenBank and accession numbers were obtained (MN719478 and MT430902). The available 16S rRNA gene sequences of Pipistrellus coromondra and Pipistrellus kuhlii lepidus were retrieved from NCBI and incorporated in N-J tree analysis. Overall, the interspecific genetic variations among Pipistrellus coromondra and Pipistrellus kuhlii lepidus were 8% and 1% respectively. In our recommendation, a comprehensive molecular identification of bats is need of hour to report more cryptic and new species from Pakistan.
Um total de 10 espécimes foi capturado em locais selecionados da Bajaur Agency FATA, Paquistão, usando redes de neblina. Os espécimes capturados foram identificados morfologicamente e várias medidas morfométricas foram realizadas. O comprimento da cabeça e do corpo (HB) de Pipistrellus coromondra e Pipistrellus kuhlii lepidus (n = 10) foi de 43 ± 0,11 mm e 45 ± 1,1, respectivamente. Pipistrellus kuhlii identificado morfologicamente e confirmado como Pipistrellus kuhlii lepidus com base em sequências de rRNA 16S. As sequências de DNA foram submetidas ao GenBank e os números de acesso foram obtidos (MN 719478 e MT430902). As sequências do gene 16S rRNA disponíveis de Pipistrellus coromondra e Pipistrellus kuhlii lepidus foram recuperadas do NCBI e incorporadas na análise da árvore N-J. No geral, as variações genéticas interespecíficas entre Pipistrellus coromondra e Pipistrellus kuhlii lepidus foram de 8% e 1%, respectivamente. Em nossa recomendação, uma identificação molecular abrangente de morcegos precisa de uma hora para relatar mais espécies crípticas e novas do Paquistão.
Asunto(s)
Animales , Quirópteros/anatomía & histología , Quirópteros/clasificación , Quirópteros/genéticaRESUMEN
Abstract A total of 10 specimens were captured from selected sites of Bajaur Agency FATA, Pakistan using mist nets. The captured specimens were morphologically identified and various morphometric measurements were taken. The head and Body length (HB) of Pipistrellus coromondra and Pipistrellus kuhlii lepidus (n=10) was 43±0.11 mm and 45±1.1 respectively. Morphologically identified Pipistrellus kuhlii confirmed as Pipistrellus kuhlii lepidus based on 16S rRNA sequences. The DNA sequences were submitted to GenBank and accession numbers were obtained (MN 719478 and MT430902). The available 16S rRNA gene sequences of Pipistrellus coromondra and Pipistrellus kuhlii lepidus were retrieved from NCBI and incorporated in N-J tree analysis. Overall, the interspecific genetic variations among Pipistrellus coromondra and Pipistrellus kuhlii lepidus were 8% and 1% respectively. In our recommendation, a comprehensive molecular identification of bats is need of hour to report more cryptic and new species from Pakistan.
Resumo Um total de 10 espécimes foi capturado em locais selecionados da Bajaur Agency FATA, Paquistão, usando redes de neblina. Os espécimes capturados foram identificados morfologicamente e várias medidas morfométricas foram realizadas. O comprimento da cabeça e do corpo (HB) de Pipistrellus coromondra e Pipistrellus kuhlii lepidus (n = 10) foi de 43 ± 0,11 mm e 45 ± 1,1, respectivamente. Pipistrellus kuhlii identificado morfologicamente e confirmado como Pipistrellus kuhlii lepidus com base em sequências de rRNA 16S. As sequências de DNA foram submetidas ao GenBank e os números de acesso foram obtidos (MN 719478 e MT430902). As sequências do gene 16S rRNA disponíveis de Pipistrellus coromondra e Pipistrellus kuhlii lepidus foram recuperadas do NCBI e incorporadas na análise da árvore N-J. No geral, as variações genéticas interespecíficas entre Pipistrellus coromondra e Pipistrellus kuhlii lepidus foram de 8% e 1%, respectivamente. Em nossa recomendação, uma identificação molecular abrangente de morcegos precisa de uma hora para relatar mais espécies crípticas e novas do Paquistão.
Asunto(s)
Animales , Quirópteros/genética , Pakistán , ARN Ribosómico 16SRESUMEN
Extensive field surveys were carried out to explore the distribution of Leislers Bat Nyctalus leisleri (Kuhl, 1819) in selected area of FATA regions, Pakistan. Specimens of Leisler's Bat Nyctalus leisleri (Kuhl, 1819) (n5) were collected from Kurram Agency (Shublan) (N33.8229788 E70.1634414) at elevation 1427m and Khyber Agency (Landi Kotel) (N34.0909899 E71.1457517) at elevation 1091m for two years survey extending from May 2013 through August 2015. The mean head and body length, hind foot length, ear length and tail length the Nyctalus leisleri specimens captured from the study area was 65.08 ± 1.58 mm, 44.06 ± 0.52 mm, 8.38 ± 0.60 mm, 13.20 ± 0.99 mm and 39.46 ± 1.46 mm, respectively. For molecular analysis the sequences of COI gene were obtained and analyzed. The mean intraspecific divergences of Nyctalus leisleri was 0.04%. The mean interspecific divergences of Nyctalus noctula and Nyctalus leisleri was 0.2%. The mean concentration of each nucleotides was A = (26.3%), T = (32.8%), G = (15.9%) and C = (25.0%). The mean A+T contents were 59.2%and C+G were 40.9%. In the phylogenetic tree Nyctalus leisleri and Nyctalus noctula clustered with significant bootstrap support value.
Extensas pesquisas de campo foram realizadas para explorar a distribuição do morcego de Leisler Nyctalus leisleri (Kuhl, 1819), em uma área selecionada das regiões das FATA, Paquistão. Espécimes do morcego de Leisler Nyctalus leisleri (Kuhl, 1819) (n = 5) foram coletados na Agência Kurram (Shublan) (N33.8229788 E70.1634414), na elevação 1.427 m, e na Agência Khyber (Landi Kotel) (N34.0909899 E71.1457517), na elevação 1.091 m, por dois anos de pesquisa, estendendo-se de maio de 2013 a agosto de 2015. Os comprimentos médios da cabeça, do corpo, do pé traseiro, da orelha e da cauda dos espécimes de Nyctalus leisleri capturados na área de estudo foram de 65,08 ± 1,58 mm, 44,06 ± 0,52 mm, 8,38 ± 0,60 mm, 13,20 ± 0,99 mm e 39,46 ± 1,46 mm, respectivamente. Para análise molecular, foram obtidas e analisadas as sequências do gene COI. A média das divergências intraespecíficas de Nyctalus leisleri foi de 0,04%. As divergências interespecíficas médias de Nyctalus noctula e Nyctalus leisleri foram de 0,2%. A concentração média de cada nucleotídeos foi A = 26,3%, T = 32,8%, G = 15,9% e C = 25%. Os conteúdos médios de A + T foram de 59,2% e de C + G foram de 40,9%. Na árvore filogenética, Nyctalus leisleri e Nyctalus noctula agruparam-se com um valor significativo de suporte de bootstrap.
Asunto(s)
Animales , Quirópteros/anatomía & histología , Quirópteros/genéticaRESUMEN
Molecular based identification of bat fauna in Pakistan has been relatively less explored. The current study was therefore planned to report for the first time the molecular classification of insectivorous bats (Pipistrellus coromandra) based on mitochondrion gene (COI) from Punjab, Pakistan. Specimens were collected from five different locations followed by DNA extraction with subsequent gene amplification and sequencing. All samples in the study had shown close identity matches with species (Pipistrellus coromandra) from India and (Pipistrellus tenuis) from Vietnam with percentage identity score of 96.11 and 95.58 respectively except one sequence which only revealed 86.78% identity match on Basic Local Alignment Search Tool (BLAST) and could only be assigned to genus level Pipistrellus sp. The results indicated negligible intra-population genetic distance among collected samples whereas the comparison with species from other countries had shown high intraspecific (P. coromandra) and interspecific (P. tenuis) mean genetic distances. The current study hence successfully proved the efficiency of COI gene as a molecular marker for species identification and in analyzing the patterns of genetic variation with species from other countries.
A identificação com base molecular da fauna de morcegos no Paquistão tem sido relativamente menos explorada. Portanto, o estudo atual foi planejado para relatar pela primeira vez a classificação molecular de morcegos insetívoros (Pipistrellus coromandra) com base no gene da mitocôndria (COI) de Punjab, Paquistão. As amostras foram coletadas em cinco locais diferentes, seguidas pela extração de DNA com subsequente amplificação e sequenciamento do gene. Todas as amostras no estudo mostraram coincidências de identidade próximas com espécies (Pipistrellus coromandra) da Índia e (Pipistrellus tenuis) do Vietnã, com pontuação de identidade percentual de 96,11 e 95,58, respectivamente, exceto uma sequência que revelou apenas 86,78% de correspondência de identidade na Ferramenta de Pesquisa de Alinhamento Local Básico (BLAST), a qual só poderia ser atribuída ao nível de gênero Pipistrellus sp. Os resultados indicaram distância genética intrapopulacional desprezível entre as amostras coletadas, enquanto a comparação com espécies de outros países mostrou altas distâncias genéticas intraespecíficas (P. coromandra) e interespecíficas (P. tenuis) médias. O presente estudo, portanto, comprovou com sucesso a eficiência do gene COI como marcador molecular para identificação de espécies e análise dos padrões de variação genética com espécies de outros países.
Asunto(s)
Animales , Mapeo Cromosómico/veterinaria , Quirópteros/genética , Marcadores GenéticosRESUMEN
Molecular based identification of bat fauna in Pakistan has been relatively less explored. The current study was therefore planned to report for the first time the molecular classification of insectivorous bats (Pipistrellus coromandra) based on mitochondrion gene (COI) from Punjab, Pakistan. Specimens were collected from five different locations followed by DNA extraction with subsequent gene amplification and sequencing. All samples in the study had shown close identity matches with species (Pipistrellus coromandra) from India and (Pipistrellus tenuis) from Vietnam with percentage identity score of 96.11 and 95.58 respectively except one sequence which only revealed 86.78% identity match on Basic Local Alignment Search Tool (BLAST) and could only be assigned to genus level Pipistrellus sp. The results indicated negligible intra-population genetic distance among collected samples whereas the comparison with species from other countries had shown high intraspecific (P. coromandra) and interspecific (P. tenuis) mean genetic distances. The current study hence successfully proved the efficiency of COI gene as a molecular marker for species identification and in analyzing the patterns of genetic variation with species from other countries.
A identificação com base molecular da fauna de morcegos no Paquistão tem sido relativamente menos explorada. Portanto, o estudo atual foi planejado para relatar pela primeira vez a classificação molecular de morcegos insetívoros (Pipistrellus coromandra) com base no gene da mitocôndria (COI) de Punjab, Paquistão. As amostras foram coletadas em cinco locais diferentes, seguidas pela extração de DNA com subsequente amplificação e sequenciamento do gene. Todas as amostras no estudo mostraram coincidências de identidade próximas com espécies (Pipistrellus coromandra) da Índia e (Pipistrellus tenuis) do Vietnã, com pontuação de identidade percentual de 96,11 e 95,58, respectivamente, exceto uma sequência que revelou apenas 86,78% de correspondência de identidade na Ferramenta de Pesquisa de Alinhamento Local Básico (BLAST), a qual só poderia ser atribuída ao nível de gênero Pipistrellus sp. Os resultados indicaram distância genética intrapopulacional desprezível entre as amostras coletadas, enquanto a comparação com espécies de outros países mostrou altas distâncias genéticas intraespecíficas (P. coromandra) e interespecíficas (P. tenuis) médias. O presente estudo, portanto, comprovou com sucesso a eficiência do gene COI como marcador molecular para identificação de espécies e análise dos padrões de variação genética com espécies de outros países.
Asunto(s)
Animales , Quirópteros/genética , Pakistán , FilogeniaRESUMEN
Abstract The flying fox (Pteropus giganteus) also familiar with the name of the greater Indian fruit Bat belongs to the order Chiroptera and family Pteropodidae. Current research emphasis on the DNA barcoding of P. giganteus in Azad Jammu Kashmir. Bat sequences were amplified and PCR products were sequenced and examined by bioinformatics software. Congeneric and conspecific, nucleotide composition and K2P nucleotide deviation, haplotype diversity and the number of haplotypes were estimated. The analysis showed that all of the five studied samples of P. giganteus had low G contents (G 19.8%) than C (27.8%), A (25.1%) and T (27.3%) contents. The calculated haplotype diversity was 0.60% and the mean intraspecific K2P distance was 0.001% having a high number of transitional substitutions. The study suggested that P. giganteus (R=0.00) do not deviate from the neutral evolution. It was determined from the conclusion that this mtDNA gene is a better marker for identification of Bat species than nuclear genes due to its distinctive characteristics and may serve as a landmark for the identification of interconnected species at the molecular level and in the determination of population genetics.
Resumo A raposa-voadora (Pteropus giganteus), também conhecida como morcego indiano, pertence à ordem dos Chiroptera e à família Pteropodidae. A presente pesquisa dá ênfase ao código de barras de DNA de P. giganteus em Azad Jammu e Caxemira. Sequências genéticas dos morcegos foram amplificadas, e os produtos de PCR foram sequenciados e examinados por software de bioinformática. De espécies congenérica e coespecífica, foram estimados composição nucleotídica e desvio de nucleotídeos K2P, diversidade de haplótipos e número de haplótipos. A análise mostrou que todas as cinco amostras estudadas de P. giganteus apresentaram baixos teores de G (19,8%) em comparação com C (27,8%), A (25,1%) e T (27,3%). A diversidade de haplótipos calculada foi de 0,60%, e a distância média intraespecífica de K2P foi de 0,001%, com um elevado número de substituições transicionais. O estudo sugeriu que P. giganteus (R = 0,00) não se desviou da evolução neutra. É possível concluir que o gene mtDNA é um marcador favorável para identificação de espécies de morcegos do que genes nucleares por causa de suas características distintivas e pode servir como um marco para a identificação de espécies interconectadas em nível molecular e para a determinação genética de populações.
Asunto(s)
Animales , Quirópteros/genética , Pakistán , Haplotipos/genética , ADN Mitocondrial , Código de Barras del ADN TaxonómicoRESUMEN
ABSTRACT Here we present detailed descriptions and comparisons of the axial skeleton of seven species of bats belonging to five subfamilies of Phyllostomidae of different trophic guilds. The material examined consisted of 34 complete skeletons of seven species. For five of the studied species, previous descriptions have not been conducted, and for the vampires only limited information is available, so that descriptions for these species are here completed. The axial skeleton has characters that allow grouping of the species phylogenetically of the same subfamily and by feeding habits. At the same time, there are characters that associate species from different subfamilies with different types of diet or ways to obtain food.
Asunto(s)
Animales , Masculino , Femenino , Huesos/anatomía & histología , Quirópteros/anatomía & histología , Filogenia , Argentina , Alas de Animales , Quirópteros/clasificación , Quirópteros/genéticaRESUMEN
Tuberculosis (TB) is one of the infectious diseases that contributes most to the morbidity and mortality of millions of people worldwide. Brazil is one of 22 countries that accounts for 80% of the tuberculosis global burden. The highest incidence rates in Brazil occur in the States of Amazonas and Rio de Janeiro. The aim of this study was to describe the temporal distribution of TB in the State of Amazonas. Between 2001 and 2011, 28,198 cases of tuberculosis were reported in Amazonas, distributed among 62 municipalities, with the capital Manaus reporting the highest (68.7%) concentration of cases. Tuberculosis was more prevalent among males (59.3%) aged 15 to 34 years old (45.5%), whose race/color was predominantly pardo (64.7%) and who had pulmonary TB (84.3%). During this period, 81 cases of multidrug-resistant TB were registered, of which the highest concentration was reported from 2008 onward (p = 0.002). The municipalities with the largest numbers of indigenous individuals affected were São Gabriel da Cachoeira (93%), Itamarati (78.1%), and Santa Isabel do Rio Negro (70.1%). The future outlook for this region includes strengthening the TB control at the primary care level, by expanding diagnostic capabilities, access to treatment, research projects developed in collaboration with the Dr. Heitor Vieira Dourado Tropical Medicine Foundation .;Fundação de Medicina Tropical Dr. Heitor Vieira Dourado (FMT-HVD).; and financing institutions, such as the project for the expansion of the Clinical Research Center and the creation of a hospital ward for individuals with transmissible respiratory diseases, including TB.
Asunto(s)
Animales , Femenino , Migración Animal , Quirópteros/genética , Micosis/transmisión , Características de la Residencia , Conservación de los Recursos Naturales , Quirópteros/microbiología , Demografía , ADN Mitocondrial/genética , Complejo IV de Transporte de Electrones/genética , Flujo Génico , Variación Genética , Genética de Población , Hibernación , Repeticiones de Microsatélite/genética , Micosis/microbiología , Pennsylvania , FilogeografíaRESUMEN
A limited number of studies have focused on the population genetic structure of vampire bats (Desmodus rotundus) in America. This medium-sized bat is distributed in tropical areas of the continent with high prevalence in forested livestock areas. The aim of this work was to characterize the vampire population structure and their genetic differentiation. For this, we followed standard methods by which live vampires (caught by mist-netting) and preserved material from scientific collections, were obtained for a total of 15 different locations, ranging from Chihuahua (North) to Quintana Roo (Southeast). Tissue samples were obtained from both live and collected animals, and the genetic differentiation, within and among localities, was assessed by the use of seven microsatellite loci. Our results showed that all loci were polymorphic and no private alleles were detected. High levels of heterozygosis were detected when the proportion of alleles in each locus were compared. Pairwise F ST and R ST detected significant genetic differentiation among individuals from different localities. Our population structure results indicate the presence of eleven clusters, with a high percentage of assigned individuals to some specific collecting site. Rev. Biol. Trop. 62 (2): 659-669. Epub 2014 June 01.
Muy pocos trabajos se han enfocado en el estudio genético de las poblaciones de vampiro (Desmodus rotundus) en América. Este murciélago de tamaño mediano se encuentra distribuido en las áreas tropicales de América, con una gran prevalencia en zonas de ganadería. La recolecta de tejidos se realizó mediante redes de niebla y en con ejemplares de colecciones, estas dan un total de 15 localidades. Mediante el uso de siete microsatellites, nosotros estudiamos la diferenciación genética dentro y entre localidades muestreadas, estas fueron desde Chihuahua en el norte, hasta Quintana Roo en el sureste. Todos los loci fueron polimórficos y no se encontraron alelos privados. Se encontraron altos niveles de heterócigos cuando se compararon la proporción de alelos en cada locus. Comparaciones pareadas de F ST y R ST mostraron una diferenciación genética entre los individuos de las diferentes localidades. Los resultados de estructura genética indican la presencia de once clusters, con un alto porcentaje de asignación de los individuos a las localidades en donde fueron recolectados.
Asunto(s)
Animales , Quirópteros/genética , Variación Genética , Repeticiones de Microsatélite/genética , Quirópteros/clasificación , Heterocigoto , MéxicoRESUMEN
Pneumocystis has been isolated from a wide range of unrelated mammalian hosts, including humans, domestic and wild animals. It has been demonstrated that the genome of Pneumocystis of one host differs markedly from that of other hosts. Also, variation in the chromosome and DNA sequence of Pneumocystis within a single host species has been observed. Since information about the occurrence and nature of infections in wild animals is still limited, the objective of this work was to detect the presence of Pneumocystis sp. in lungs of bats from two states from Brazil by Nested-PCR amplification. The bats, captured in caves and in urban areas, were obtained from the Program of Rabies Control of two States in Brazil, Mato Grosso and Rio Grande do Sul, located in the Mid-Western and Southern regions of the country, respectively. DNAs were extracted from 102 lung tissues and screened for Pneumocystis by nested PCR at the mtLSU rRNA gene and small subunit of mitochondrial ribosomal RNA (mtSSU rRNA). Gene amplification was performed using the mtLSU rRNA, the primer set pAZ102H - pAZ102E and pAZ102X - pAZY, and the mtSSU rRNA primer set pAZ102 10FRI - pAZ102 10R-RI and pAZ102 13RI - pAZ102 14RI. The most frequent bats were Tadarida brasiliensis (25), Desmodus rotundus (20), and Nyctinomops laticaudatus (19). Pneumocystis was more prevalent in the species Nyctinomops laticaudatus (26.3 percent = 5/19), Tadarida brasiliensis (24 percent = 6/25), and Desmodus rotundus (20 percent = 4/20). Besides these species, Pneumocystis also was detected in lungs from Molossus molossus (1/11, 9.1 percent), Artibeus fimbriatus (1/1, 100 percent), Sturnira lilium (1/3, 33.3 percent), Myotis levis (2/3, 66.7 percent)and Diphylla ecaudata (1/2, 50 percent). PCR products which could indicate the presence of Pneumocystis (21.56 percent) were identified in DNA samples obtained from 8 out of 16 classified species from both states (5 bats were not identified). This is the ...
Pneumocystis tem sido isolado de uma grande variedade de hospedeiros mamíferos, incluindo humanos, animais domésticos e selvagens. Tem se demonstrado que o genoma do Pneumocystis de um hospedeiro difere marcadamente do de outros, assim como há variação no cromossomo e na seqüência de DNA dentro de uma única espécie de hospedeiro. Sabendo que a informação da ocorrência e natureza da infecção em animais silvestres ainda é limitada, o objetivo do trabalho foi detectar, por Nested-PCR, a presença de Pneumocystis sp. em pulmões de diferentes espécies de morcegos de dois estados do Brasil. Estes mamíferos voadores foram capturados em cavernas, áreas florestadas, de campo e urbanas pelo Programa de Controle da Raiva do Mato Grosso (região Centro-Oeste) e do Instituto de Pesquisas Veterinárias Desidério Finamor (RS) e Instituto Sauver no Rio Grande do Sul (região Sul). Os DNAs foram extraídos de 102 pulmões e realizado Nested-PCR utilizando os primers pAZ102H-pAZ102E e pAZ102X/R1-pAZY/R1 para amplificação do gene mtLSU-rRNA, e pAZ102 10F-RI - pAZ102 10R-RI e pAZ102 13-RI - pAZ14-RI para amplificação do gene mtSSU-rRNA. As espécies mais freqüentes foram Tadarida brasiliensis (25), Desmodus rotundus (20) e Nyctinomops laticaudatus (19). Pneumocystis foi detectado com maior prevalência nas Nyctinomops laticaudatus (26,3 por cento = 5/19), Tadarida brasiliensis (24 por cento = 6/25) e Desmodus rotundus (20 por cento = 4/20). Além destas espécies, Pneumocystis foi também detectado nos pulmões de Molossus molossus (1/11, 9,1 por cento), Artibeus fimbriatus (1/1, 100 por cento), Sturnira lilium (1/3, 33 por cento), Myotis levis (2/3, 66,7 por cento)e Diphylla ecaudata (1/2, 50 por cento). Os produtos de PCR indicaram a presença de Pneumocystis (21.56 por cento) em amostras obtidas de 8 das 16 espécies classificadas para ambos os estados (cinco morcegos não foram classificados). Este é o primeiro registro de detecção de Pneumocystis em morcegos no Brasil.
Asunto(s)
Animales , Pneumocystis/aislamiento & purificación , Pulmón/microbiología , Quirópteros/genética , Quirópteros/microbiología , Reacción en Cadena de la Polimerasa/métodos , Brasil , Quirópteros/clasificaciónRESUMEN
The intestines of Sturnira lilium, Phyllostomus hastatus, Carolliaperspicillata, Glossophagasoricinaíind Desmodus rotundus were analized through macroscopic, morphometric and light microscopy studies. The species studied presented some common charactenstics generally found in bats, such as the absence of a caecum and an appendix. The frugivorous bat S. lilium has a longer intestine than the other species. The intestinal epithelium is of the simple columnar type, constituted of absorptive cells characterized by microvilli and goblet cells, which in all the species proved to be PAS + and AB +. These cells presented some differences in quantity and in distribution among the species. The intestinal mucosa presented morphologic and morphometric differences, such as the shape and size of villi as well as the length of the crypts of Lieberkühn. Histologically the different intestinal portions could be distinguished: in the middle portion we found few goblet cells and long villi; in the distal portion we found an increase in goblet cells and a decrease in villi height; and in the large intestine no villi were found, but rather abundant intestinal glands with numerous goblet cells. In C. perspicillata, P. hastatus and G. soricina we could observe Peyeras patches in the distal portion near the large intestine, whereas in D. rotundus and S. lilium we found aggregations of lymphoid nodulous tissue distributed along the tube. In all species we observed the presence of Paneth cells at the base of the crypts of Lieberkühn. We could observe differences among the species in the distribution of collagen in the small intestine submucosa and serosa. The outer muscular layer has one circular layer and a longitudinal layer, the circular layer increasing in thickness along the tube. Large intestine charactenstics are uniform among the species. Sfractures such as the taenia coli and the appendices epiploicae were absent. These results should contribute not only...
Los intestinos de Sturnira azucenas, Phyllostomus hastatus, Carollia perspicillata, Glossophaga soricina y Desmodus rotundus se analizaron a través de estudios macroscópicos, morfométricos y de microscopía de luz. Las especies estudiadas presentan algunas características comunes en general encontradas en murciélagos, tales como la ausencia de un ciego y un apéndice. Murciélagos frugívoros S. azucenas tienen un intestino más largo que el de otras especies. El epitelio intestinal es de tipo columnar simple, constituido por células de absorción que se caracterizan por microveilosidades y células caliciformes, que en todas las especies resultó ser PAS + y AB +. Estas células presentan algunas diferencias en cantidad y en la distribución entre las especies. La mucosa intestinal presenta diferencias morfológicas y morfométricas, como la forma y el tamaño de las vellosidades, así como la longitud de las criptas de Lieberkühn. Histológicamente las diferentes partes intestinales pueden ser distinguidas: en la parte media encontramos pocas células caliciformes y largas vellosidades, en la porción distal encontramos un aumento de células caliciformes y una disminución en la altura de vellosidades, y en el intestino grueso no se encontraron vellosidades, sino más bien abundantes glándulas intestinales con numerosas células caliciformes. En C. perspicillata, P hastatus y G. soricina podemos observar placas de Peyer en la porción distal, cerca del intestino grueso, mientras que en D. rotundus y S. azucenas se encontraron agrupaciones de tejido linfoide nodular distribuido a lo largo del tubo. En todas las especies se observó la presencia de células Paneth en la base de las criptas de Lieberkühn. Hemos podido observar diferencias entre las especies en la distribución de colágeno en el intestino delgado y submucosa serosa. La capa muscular exterior tiene una capa circular y una capa longitudinal, la capa circular en el aumento de espesor a lo largo del...
Asunto(s)
Masculino , Adulto , Animales , Intestinos/anatomía & histología , Intestinos/embriología , Intestinos/ultraestructura , Quirópteros/anatomía & histología , Quirópteros/fisiología , Quirópteros/genética , Microscopía Electrónica/métodos , Tracto Gastrointestinal/anatomía & histología , Tracto Gastrointestinal/ultraestructuraRESUMEN
This study verified the comparative histomorphometric adaptations in the stomach of rat, bat and pangolin in relation to diet. Ten rats, ten bats and ten pangolins of both sexes were used for this investigation. The animals were sacrificed after slight anesthesia under chloroform inhalation. The stomach were excised, fixed in 10 percent formol saline and processed for light microscopic study. Stained slides were also subjected to morphometric analysis at a magnification of 400x. The results revealed that the cellular diameter/ density of parietal and zymogenic cells are significantly different in the three mammals (p<0.05) with the exception of the diameter of the zymogenic cells in pangolin which was not statistically significant (p>0.05) when compared with that of rat. Also, histological analysis revealed slight differences in the pattern of organization and distribution of connective tissue fibers. All these observations were reflections of the different pattern the stomachs of the three mammals have adopted to cope with their respective diets.
En este estudio se verificaron las adaptaciones histomorfométricas comparativas en el estómago de ratas, murciélagos y pangolines en relación a la dieta. Se utilizaron para esta investigación 10 ejemplares de cada especie, de ambos sexos. Los animales fueron sacrificados después de anestesia bajo inhalación de cloroformo. Los estómagos fueron extirpados, fijados en formol al 10 por ciento de solución salina y procesados para su estudio microscópico de luz. Los cortes teñidos fueron también objeto de análisis morfométrico con un aumento de X 400. Los resultados revelaron que el diámetro/densidad celular de parietal y las células cimógenas son significativamente diferentes en los tres mamíferos (p <0,05), con la excepción del diámetro de la células cimógenas de pangolines que no era estadísticamente significativa (p> 0,05) en comparación con la de rata. Por otra parte, el análisis histológico reveló ligeras diferencias en las características de organización y distribución de las fibras del tejido conjuntivo. Todas estas observaciones son un reflejo del patrón de los diferentes estómagos de los tres mamíferos, que han adoptado para hacer frente a sus respectivas dietas.
Asunto(s)
Masculino , Adulto , Animales , Femenino , Estómago/anatomía & histología , Estómago/citología , Estómago/ultraestructura , Células del Tejido Conectivo/ultraestructura , Histología Comparada/métodos , Mamíferos/anatomía & histología , Mamíferos/genética , Mamíferos/metabolismo , Quirópteros/anatomía & histología , Quirópteros/fisiología , Quirópteros/genética , Ratas/anatomía & histología , Ratas/fisiologíaRESUMEN
To establish a technique which minimized the effects of fixation on the extraction of DNA from formalin-fixed tissues preserved in scientific collections we extracted DNA samples from fixed tissues using different methods and evaluated the effect of the different procedures on PCR and sequencing analysis. We investigated muscle and liver tissues from museum specimens of five species of fruit-eating (frugivorous) bats of the Neotropical genus Artibeus (Chiroptera, Phyllostomidae): A. fimbriatus, A. lituratus, A. jamaicensis, A. obscurus, and A. planirostris. The results indicated that treatment of tissues in buffered solutions at neutral pH and about 37 °C for at least four days improves the quality and quantity of extracted DNA and the quality of the amplification and sequencing products. However, the comparison between the performance of DNA obtained from fixed and fresh tissues showed that, in spite of the fact that both types of tissue generate reliable sequences for use in phylogenetic analyses, DNA samples from fixed tissues presented a larger rate of errors in the different stages of the study. These results suggest that DNA extracted from formalin-fixed tissue can be used in molecular studies of Neotropical Artibeus bats and that our methodology may be applicable to other animal groups.
Asunto(s)
Animales , Secuencia de Bases , Bancos de Muestras Biológicas , Quirópteros/genética , Filogenia , Reacción en Cadena de la PolimerasaRESUMEN
Random amplified polymorphic DNA molecular marker was utilized as a means of analyzing genetic variability in seven bat species: Molossus molossus, M. rufus, Eumops glaucinus, E. perotis, Myotis nigricans, Eptesicus furinalis, and Artibeus planirostris. The determination of genetic diversity was based on 741 bands produced by a 20-random primer set. Only eight bands were considered monomorphic to one species. The greatest number of bands and the most polymorphic condition were exhibited by M. molossus, followed by M. nigricans, A. planirostris, E. furinalis, E. glaucinus, M. rufus, and E. perotis. Nei's genetic diversity index in the seven species considering the 20 primers was not greater than 0.22, but some primers were capable of detecting values between 0.39 and 0.49. Nei's unbiased genetic distance values and the UPGMA clustering pattern show that M. molossus and M. rufus have a close genetic relationship, unlike that observed between E. perotis and E. glaucinus. The latter was clustered with A. planirostris and E. furinalis. The low values for genetic diversity and distance observed indicate a genetic conservatism in the seven species. The fluorescent in situ hybridization experiments did not confirm a monomorphic condition for the eight bands identified, demonstrating that the monomorphic bands obtained by random amplified polymorphic DNA are insufficient for the identification of bat species.
Asunto(s)
Animales , Variación Genética , Quirópteros/genética , Técnica del ADN Polimorfo Amplificado Aleatorio , Hibridación Fluorescente in Situ , Marcadores Genéticos/genética , Quirópteros/clasificación , Reproducibilidad de los ResultadosRESUMEN
The genus Uroderma includes two species: U. magnirostrum and U. bilobatum. These species are characterized by their high degree of karyotypic evolution, diverging from most other species of the subfamily Stenodermatinae, which have a lower degree of chromosomic evolution. The present study reports the first banding patterns of U. magnirostrum (G-, C-banding and Ag-NOR) and U. bilobatum (C-banding and Ag-NOR). The chromosomic data in conventional staining of U. magnirostrum (2n = 36, NF = 62) and U. bilobatum (cytotype 2n = 42, NF = 50) are equivalent to that described in the literature. When compared, chromosomal homeologies are found in both karyotypes, as well as differences, confirming that karyotypic evolution in the Uroderma genus is intense. Fission, fusion, inversion or translocation events are required to explain the karyotypic evolution of this genus. The comparison of karyotype, described here, to one of the species of the genus Artibeus (2n = 30/31), suggests that some chromosomic forms are apomorphic and shared between the two species of Uroderma. This confirms the monophyly of the genus, and that U. magnirostrum presents a more primitive karyotype when compared to U. bilobatum.
Asunto(s)
Animales , Quirópteros/genética , Análisis Citogenético , Brasil , CariotipificaciónRESUMEN
The purpose of this work was to verify the ability of the enzyme Alu I to cleave and/or remove satellite DNA sequences from heterochromatic regions in chromosomes of bats, by identifying the occurrence of modifications in the pattern of fluorescence in situ hybridization with telomeric DNA. The localization and fluorescence intensity of the telomeric DNA sites of the Alu-digested and undigested chromosomes of species Eumops glaucinus, Carollia perspicillata, and Platyrrhinus lineatus were analyzed. Telomeric sequences were detected at the termini of chromosomes of all three species, although, in C. perspicillata, the signals were very faint or absent in most chromosomes. This finding was interpreted as being due to a reduced number of copies of the telomeric repeat, resulting from extensive telomeric association and/or rearrangements undergone by the chromosomes of Carollia. Fluorescent signals were also observed in centromeric and pericentromeric regions in several two-arm chromosomes of E. glaucinus and C. perspicillata. In E. glaucinus and P. lineatus, some interstitial and terminal telomeric sites were observed to be in association with regions of constitutive heterochromatin and ribosomal DNA (NORs). After digestion, these telomeric sites showed a significant decrease in signal intensity, indicating that enzyme Alu I cleaves and/or removes part of the satellite DNA present in these regions. These results suggest that the telomeric sequence is a component of the heterochromatin, and that the C-band- positive regions of bat chromosomes have a different DNA composition
Asunto(s)
Humanos , Animales , Secuencia de Bases , Cromosomas Humanos , Hibridación Fluorescente in Situ , Quirópteros/genética , Telómero/genética , Bandeo CromosómicoRESUMEN
We developed a procedure to obtain fibroblasts from bat skin. A small fragment of the ear is removed under ether anesthesia. This material is then cut up into small pieces and cultured in standard cell culture media. Very good quality chromosome preparations for cytogenetic studies are obtained in about three weeks. Secondary cultures can be used for other biological studies. This procedure does not require sacrificing the animals
Asunto(s)
Animales , Fibroblastos , Piel/citología , Quirópteros/genética , Biopsia , Análisis CitogenéticoRESUMEN
A staining mixture of hematoxylin-iron alum combined with a strong hydrochloric hydrolysis was successfully applied for chromosome observation of several kinds of plants and some animals. Slightly different procedures were developed for different materials and objectives. For plant cells, the most important technical aspect was the use of 5 N HCl hydrolysis, which resulted in a very transparent cytoplasm, combined with an intense, specific hematoxylin stain. This technique is recommended for cytogenetical analysis in general, and it is especially indicated for practical classes, due to its simplicity and high reproducibility of results. Moreover, the deep contrast observed makes this technique very useful for sequential staining of cells previously analyzed with other stains, as well as for materials with fixation problems.
Asunto(s)
Animales , Quirópteros , Cromosomas/química , Colorantes , Hematoxilina , Insectos/citología , Plantas/citología , Hidrólisis , Insectos/genética , Meiosis , Mitosis , Plantas/genética , Quirópteros/genéticaRESUMEN
O presente trabalho apresenta dados cromossômicos de cinco espécies de morcegos da subfamília Stenodermatinae. Os resultados pertencentes as espécies Artibeus planirostris, A. lituratus, A. cinereus, Sturnira lilium e Vampyrops lineatus säo comparados. Grande similaridades nos padröes de bandas G e significantes diferenças na localizaçäo das regiöes organizadoras de nucléolos e no padräo de heterocromatina constitutiva foram encontradas entre as cinco espécies estudadas