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BACKGROUND Panstrongylus rufotuberculatus (Hemiptera-Reduviidae) is a triatomine species with a wide geographic distribution and a broad phenotypic variability. In some countries, this species is found infesting and colonising domiciliary ecotopes representing an epidemiological risk factor as a vector of Trypanosoma cruzi, etiological agent of Chagas disease. In spite of this, little is known about P. rufotuberculatus genetic diversity. METHODS Cytogenetic studies and DNA sequence analyses of one nuclear (ITS-2) and two mitochondrial DNA sequences (cyt b and coI) were carried out in P. rufotuberculatus individuals collected in Bolivia, Colombia, Ecuador and Mexico. Moreover, a geometric morphometrics study was applied to Bolivian, Colombian, Ecuadorian and French Guiana samples. OBJECTIVES To explore the genetic and phenetic diversity of P. rufotuberculatus from different countries, combining chromosomal studies, DNA sequence analyses and geometric morphometric comparisons. FINDINGS We found two chromosomal groups differentiated by the number of X chromosomes and the chromosomal position of the ribosomal DNA clusters. In concordance, two main morphometric profiles were detected, clearly separating the Bolivian sample from the other ones. Phylogenetic DNA analyses showed that both chromosomal groups were closely related to each other and clearly separated from the remaining Panstrongylus species. High nucleotide divergence of cyt b and coI fragments were observed among P. rufotuberculatus samples from Bolivia, Colombia, Ecuador and Mexico (Kimura 2-parameter distances higher than 9%). MAIN CONCLUSIONS Chromosomal and molecular analyses supported that the two chromosomal groups could represent different closely related species. We propose that Bolivian individuals constitute a new Panstrongylus species, being necessary a detailed morphological study for its formal description. The clear morphometric discrimination based on the wing venation pattern suggests such morphological description might be conclusive.
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Abstract Genetic and phylogenetic relationships among seven piranha species of the genera Serrasalmus and Pygocentrus from the Paraná-Paraguay, São Francisco and Tocantins River basins were evaluated in the present study by partial sequences of two mitochondrial genes, Cytochrome b and Cytochrome c Oxidase I. Phylogenetic analysis of Maximum-Likelihood and Bayesian inference were performed. Results indicated, in general, greater genetic similarity between the two species of Pygocentrus (P. nattereri and P. piraya), between Serrasalmus rhombeus and S. marginatus and between S. maculatus, S. brandtii and S. eigenmanni. Pygocentrus nattereri, S. rhombeus and S. maculatus showed high intraspecific genetic variability. These species have each one, at least two different mitochondrial lineages that, currently, occur in sympatry (S. rhombeus) or in allopatry (P. nattereri and S. maculatus). Species delimitation analysis and the high values of genetic distances observed between populations of S. rhombeus and of S. maculatus indicated that each species may corresponds to a complex of cryptic species. The non-monophyletic condition of S. rhombeus and S. maculatus reinforces the hypothesis. The geographic distribution and the genetic differentiation pattern observed for the piranha species analyzed herein are discussed regarding the geological and hydrological events that occurred in the hydrographic basins.
Resumo Relações genéticas e filogenéticas de sete espécies de piranhas dos gêneros Serrasalmus e Pygocentrus das bacias hidrográficas Paraná-Paraguai, São Francisco e Tocantins foram avaliadas com base em sequências parciais dos genes mitocondriais Citocromo b e Citocromo c Oxidase I. Foram realizadas análises filogenéticas de Máxima Verossimilhança e de inferência Bayesiana. Os resultados indicaram, em geral, maior similaridade genética entre as duas espécies de Pygocentrus (P. nattereri e P. piraya), entre Serrasalmus rhombeus e S. marginatus e entre S. maculatus, S. brandtii e S. eigenmanni. Pygocentrus nattereri, S. rhombeus e S. maculatus revelaram ter alta variabilidade genética intraespecífica. Essas espécies têm, cada uma, pelo menos duas linhagens mitocondriais que, atualmente, ocorrem em simpatria (S. rhombeus) ou alopatria (P. nattereri e S. maculatus). Análises de delimitação de espécies e os altos valores de distância genética observados entre as populações de S. rhombeus e de S. maculatus indicam que cada espécie pode, na verdade, corresponder a um complexo de espécies crípticas. A condição não-monofilética de S. rhombeus e S. maculatus reforça essa hipótese. A distribuição geográfica e o padrão de diferenciação genética observados para as espécies de piranhas analisadas são discutidos com relação aos eventos geológicos e hidrológicos que ocorreram nas bacias hidrográficas.
Subject(s)
Animals , Characiformes , Paraguay , Phylogeny , Brazil , Bayes Theorem , RiversABSTRACT
Objective To analyze the sequences of the cytochrome C oxidase subunit I (Cox1) gene of various Echinococcus granulosus genotypes that are currently recorded in the GenBank database, so as to investigate the genetic variation and differentiation of the E. granulosus genotypes across the world. Methods The sequences of the Cox1 gene of various E. granulosus genotypes that are currently recorded in the GenBank database were collected, and the same sequences of the Cox1 gene identified from a region were excluded. The mutation sites among the Cox1 gene sequences were identified and a phylogenetic tree was created based on the Cox1 gene. Results Transversion mutation was the predominant type of mutation in the Cox1 gene of E. granulosus. The same Cox1 gene sequence was found in E. granulosus G1, G6 and G7 genotypes isolated from various geographical locations across the world, with the corresponding GenBank accession numbers of KY766891, MH300971 and MH301007, respectively. Phylogenetic analysis revealed that E. granulosus G10 genotype had a remarkable geographical aggregation. Conclusions E. granulosus G1, G6 and G7 genotypes have primitive Cox1 gene sequences. There is a geographical aggregation of the E. granulosus G10 genotype in the phylogenetic tree, which has a tendency towards reproductive isolation.
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Brachionus quadridentatus es una especie morfológicamente variable, distribuida por todo el mundo. Su taxonomía es confusa debido a las numerosas variantes infrasubespecíficas descritas en este taxón. Con la taxonomía basada en la morfología, B. quadridentatus tiene tres variantes reconocidas: B. quadridentatus quadridentatus, B. quadridentatus f. brevispinus and B. quadridentatus f. cluniorbicularis. En este estudio, exploramos la diversidad genética entre algunas poblaciones de B. quadridentatus, usando secuencias de los genes COI ADNmt y 18S ARNr. El análisis de delimitación de especies coalescente usando el gen 18S apoya la presencia de al menos tres especies putativas dentro del complejo B. quadridentatus. Estos resultados estuvieron en concordancia con los análisis filogenético y GMYC usando el gen 18S. Sin embargo, se encontró variación en morfología y secuencias del gen COI dentro de cada una de las tres especies putativas. Se encontraron siete linajes delimitados por las secuencias del gen COI usando el método de delimitación ABGD, que además están morfologicamente diferenciadas. Se encontró discordancia mitonuclear entre la filogenia del gen COI y la del gen 18S. La incongruencia entre el marcador mitocondrial y el nuclear puede ser explicada por sorteo incompleto de linaje.
Brachionus quadridentatus is a morphologically variable species of rotifer distributed worldwide. The taxonomy of this species is confused, with numerous infrasubspecific variants described in the taxon: B. quadridentatus quadridentatus, B. quadridentatus f. brevispinus and B. quadridentatus f. cluniorbicularis. In this study, we explored genetic diversity among some populations of B. quadridentatus, using sequences of mitochondrial COI and nuclear 18S genes. The coalescent species delimitation analysis with the 18S gene highly supports the presence of at least three putative species within the B. quadridentatus complex. These results were in agreement with the phylogenetic and GMYC analysis using the 18S gene. However, we also found variation within each of these three putative species in morphology and COI gene sequences. There were seven morphologically differentiated lineages that were recovered as distinct based on COI gene sequences using the ABGD delimitation method. As such, mitonuclear discordance between COI and 18S phylogenies was found. The incongruence between mitochondrial and nuclear markers could be explained by incomplete lineage sorting.
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Avian malaria is one of the most important general blood parasites of poultry in Southeast Asia. Plasmodium (P.) juxtanucleare causes avian malaria in wild and domestic fowl. This study aimed to identify and characterize the Plasmodium species infecting in Thai native fowl. Blood samples were collected for microscopic examination, followed by detection of the Plasmodium cox I gene by using PCR. Five of the 10 sampled fowl had the desired 588 base pair amplicons. Sequence analysis of the five amplicons indicated that the nucleotide and amino acid sequences were homologous to each other and were closely related (100% identity) to a P. juxtanucleare strain isolated in Japan (AB250415). Furthermore, the phylogenetic tree of the cox I gene showed that the P. juxtanucleare in this study were grouped together and clustered with the Japan strain. The presence of P. juxtanucleare described in this study is the first report of P. juxtanucleare in the Thai native fowl of Thailand.
Subject(s)
Animals , Humans , Amino Acid Sequence , Asia, Southeastern , Asian People , Base Pairing , Cytochromes c , Cytochromes , Electron Transport Complex IV , Japan , Malaria, Avian , Parasites , Plasmodium , Polymerase Chain Reaction , Poultry , Sequence Analysis , Thailand , TreesABSTRACT
Un total de 12 cestodos adultos se colectaron de los conductos biliares de ratones domésticos (Mus musculus) provenientes de Lima, Perú. Diversas características del escólex y proglotis maduros del cestodo fueron observadas para la identificación morfológica. Así mismo, se realizó un diagnóstico molecular mediante un PCR y secuenciación parcial del gen mitocondrial citocromo c oxidasa subunidad 1 (cox1). Todos los cestodos fueron identificados como Hymenolepis microstoma por morfología y métodos moleculares. El aislado de H. microstoma de Perú mostró una similitud de secuencia significativa (> 99%) con los aislados de H. microstoma previamente reportados. Nuestro informe confirma la presencia del parásito en ratones de Lima.
A total of 12 adult cestodes were collected from the bile ducts of domestic mice (Mus musculus) from Lima, Peru. Various features of the scolex and mature proglottids of the tapeworms were observed for morphological identification. A molecular diagnosis was performed by PCR-based partial sequencing of mitochondrial gene of cytochrome c oxidase subunit 1 (cox1). All cestodes were identified as Hymenolepis microstoma by morphology and molecular methods. The H. microstoma isolate from Peru showed significant sequence similarity with previously reported isolates of H. microstoma (>99%). Our report confirms the presence of the parasite in mice from Lima.
ABSTRACT
Abstract Genetic and phylogenetic relationships among seven piranha species of the genera Serrasalmus and Pygocentrus from the Paraná-Paraguay, São Francisco and Tocantins River basins were evaluated in the present study by partial sequences of two mitochondrial genes, Cytochrome b and Cytochrome c Oxidase I. Phylogenetic analysis of Maximum-Likelihood and Bayesian inference were performed. Results indicated, in general, greater genetic similarity between the two species of Pygocentrus (P. nattereri and P. piraya), between Serrasalmus rhombeus and S. marginatus and between S. maculatus, S. brandtii and S. eigenmanni. Pygocentrus nattereri, S. rhombeus and S. maculatus showed high intraspecific genetic variability. These species have each one, at least two different mitochondrial lineages that, currently, occur in sympatry (S. rhombeus) or in allopatry (P. nattereri and S. maculatus). Species delimitation analysis and the high values of genetic distances observed between populations of S. rhombeus and of S. maculatus indicated that each species may corresponds to a complex of cryptic species. The non-monophyletic condition of S. rhombeus and S. maculatus reinforces the hypothesis. The geographic distribution and the genetic differentiation pattern observed for the piranha species analyzed herein are discussed regarding the geological and hydrological events that occurred in the hydrographic basins.
Resumo Relações genéticas e filogenéticas de sete espécies de piranhas dos gêneros Serrasalmus e Pygocentrus das bacias hidrográficas Paraná-Paraguai, São Francisco e Tocantins foram avaliadas com base em sequências parciais dos genes mitocondriais Citocromo b e Citocromo c Oxidase I. Foram realizadas análises filogenéticas de Máxima Verossimilhança e de inferência Bayesiana. Os resultados indicaram, em geral, maior similaridade genética entre as duas espécies de Pygocentrus (P. nattereri e P. piraya), entre Serrasalmus rhombeus e S. marginatus e entre S. maculatus, S. brandtii e S. eigenmanni. Pygocentrus nattereri, S. rhombeus e S. maculatus revelaram ter alta variabilidade genética intraespecífica. Essas espécies têm, cada uma, pelo menos duas linhagens mitocondriais que, atualmente, ocorrem em simpatria (S. rhombeus) ou alopatria (P. nattereri e S. maculatus). Análises de delimitação de espécies e os altos valores de distância genética observados entre as populações de S. rhombeus e de S. maculatus indicam que cada espécie pode, na verdade, corresponder a um complexo de espécies crípticas. A condição não-monofilética de S. rhombeus e S. maculatus reforça essa hipótese. A distribuição geográfica e o padrão de diferenciação genética observados para as espécies de piranhas analisadas são discutidos com relação aos eventos geológicos e hidrológicos que ocorreram nas bacias hidrográficas.
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Echinococcus Granulosus (EG) is the major cause of cystic echinococcosis in humans and livestock in the world. In Chile is a zoonosis of great importance. The most frequently affected geographic areas are the Regions of Aysén, Los Rios, Los Lagos, Coquimbo and the Araucanía. Hence, it was discovered that in endemic areas of hydatidosis there could be several strains and genotypes of EG. In addition, there is evidence that some strains and genotypes are more infectious for human beings than others. This interesting phenomenon of the biology of EG has been studied using molecular biology techniques based on polymerase chain reaction (PCR) and DNA sequence analysis, which has made it possible to characterize the cestode species complex called EG sensu lato (s l) as being comprised of EG sensu stricto (s.s.) (Genotypes G1-G3), E. equinus (G4), E. ortleppi (G5) and E. canadensis (G6-G10), which present an important phenotypic variation detectable in characteristics of the biological cycle, specificity of the intermediate host, pattern of development, pathogenicity, antigenicity, transmission dynamics and, consequently, in the measures needed to control the disease. The aim of this manuscript is to describe the different genotypes of EG described in humans and different livestock host reported in the literature.
Echinococcus granulosus (EG) es la principal causa de equinococosis quística en humanos y ganado en el mundo. En Chile hay una zoonosis de gran importancia. Las zonas geográficas más afectadas son las Regiones de Aysén, Los Ríos, Los Lagos, Coquimbo y la Araucanía. Por lo tanto, se descubrió que en áreas endémicas de hidatidosis podría haber varias cepas y genotipos de EG. Además, hay pruebas de que algunas cepas y genotipos son más infecciosos para los seres humanos que otros. Este interesante fenómeno de la biología del EG ha sido estudiado utilizando técnicas de biología molecular basadas en la reacción en cadena de la polimerasa (PCR) y análisis de secuencias de ADN, lo que ha permitido caracterizar el complejo de cestode llamado EG sensu lato (sl) EG (G3) y E. canadensis (G6-G10), que presentan una importante variación fenotípica detectable en las características del ciclo biológico, especificidad del huésped intermedio, patrón de desarrollo, patogenicidad, antigenicidad, dinámica de transmisión y, por consiguiente, en las medidas necesarias para el control de la enfermedad. El objetivo de este manuscrito fue describir los diferentes genotipos de EG descritos en humanos y diferentes animales de ganado reportados en la literatura.
Subject(s)
Humans , Animals , Echinococcus granulosus/genetics , Echinococcosis/parasitology , Echinococcosis/veterinary , Echinococcus granulosus/classification , Electron Transport Complex IV/genetics , Genotype , Livestock/parasitology , Molecular Typing , Random Amplified Polymorphic DNA Technique , Species SpecificityABSTRACT
Objective To apply DNA barcoding technology for exploring its taxonomic status and differences in the molecular biology of Cricetulus barabensis in Shaanxi Province.Methods Sixty-five samples of Cricetulus barabensis were collected from Dingbian,Jingbian Counties in northern of Shaanxi and Dali County in Guanzhong plain (Dingbian 58 samples,Jingbian 2 samples,and Dali 5 samples).According to the mitochondrial cytochrome C oxidase subunit I gene (CO I) sequence,the genetic distance was calculated and Neighbor-Joining tree was constructed.Results The genetic distance between two samples (13.16,13.21) and other 56 samples of Dingbian was 9.2％-10.0％.The genetic distance between the 56 samples of Dingbian and Jingbian was less than 1％ and Dali was 7.2％-8.3％;the average intraspecific genetic distance of Jingbian and Dali was less than 1％.The Neighbor-Joining tree showed that all the Cricetulus barabensis samples from the three counties were separated into two large branches.The samples of 13.16,13.21 from Dingbian together were classified into a class and the rest of the samples into another separate branch.At the same time,other samples from Dingbian except 13.16,13.21 and Jingbian were distributed in a small branch,and Dali samples were occupied another small branch.Conclusion Using the DNA barcoding technology,we have determined three subspecies of Cricetulus barabensis in Shaanxi Province,Dingbian has two kinds and Dali has a different subspecies.
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OBJECTIVE: To explore the characteristic of Chinese herbal medicine Testudinis carapax et Plastrum by the multiple PCR technique. METHODS: The salting out method was modified to extract mitochondrial DNA (mtDNA) from Testudinis carapax et Plastrum and their counterfeits. The sequences of cytochrome b (Cyt b) and cytochrome c oxidase subunit I (Col) genes were downloaded from the GenBank database, and two specific pairs of primers were designed with Premier 5.0. The genes of Testudinis carapax et Plastrum were amplified with multiple PCR and sequenced. RESULTS: The size of mtDNA got by salting out method was 16.6×103 bp, and two bright stripes between 300-500 bp were shown in agarose gel electrophoresis of authentic Testudinis carapax et Plastrum, only one or no stripe appeared for the counterfeits. CONCLUSION: The multiple PCR technique is specific, simple, and accurate for differentiation of authentic Testudinis carapax et Plastrum from their counterfeits.
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Objective: To discuss the feasibility of using partial sequence of mitochondrial cytochrome C oxidase subunit I (COI) gene as DNA barcoding to identify Saigae Tataricae Cornu (STC) in Chinese materia medica (CMM) preparation. Methods: The DNA barcoding identification was constructed in STC samples from eight different manufactories. The genome DNA was extracted by DNA extraction kit from adequately ground samples. The COI nucleotide sequences were PCR amplified with the Primers LCO1490 and HC02198 or specified primer 0703 and sequenced bi-directionally. The BLAST comparison was made in GeneBank, and the phylogenetic trees were constructed with the homologisation tree (DNAMAN) and Neighbor-Joining (NJ, MEGA 5.2) method. Saiga tatarica (gb|JN632700.1) and STC were selected as reference medicinal materials and the genetic distance was calculated using Kimura-2-parameter. Results: The lengths of partial mitochondrial COI gene collected from STC in eight samples of CMM preparation were about 658 bp. The phylogenetic tree showed that the samples of CMM preparation and reference substance assembled distinctly, and were completely separated from the outgroup. The intraspecific genetic distances of these samples ranged in 0-0.064 with an average of 0.020, and the interspecific genetic distances ranged in 0.167-0.195. Conclusion: The mitochondrial COI gene is a valid DNA barcoding gene for STC identification in CMM preparation. But other organs in antelope also contain the same gene. In order to ensure the accuracy of the results, other texst methods also need to be used cooperatively.
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To assess differentiation and relationships between Anopheles lesteri and Anopheles paraliae we established three and five iso-female lines of An. lesteri from Korea and An. paraliae from Thailand, respectively. These isolines were used to investigate the genetic relationships between the two taxa by crossing experiments and by comparing DNA sequences of ribosomal DNA second internal transcribed spacer (ITS2) and mitochondrial DNA cytochrome c oxidase subunit I (COI) and subunit II (COII). Results of reciprocal and F1-hybrid crosses between An. lesteri and An. paraliae indicated that they were compatible genetically producing viable progenies and complete synaptic salivary gland polytene chromosomes without inversion loops in all chromosome arms. The pairwise genetic distances of ITS2, COI and COII between these morphological species were 0.040, 0.007-0.017 and 0.008-0.011, respectively. The specific species status of An. paraliae in Thailand and/or other parts of the continent are discussed.
Subject(s)
Animals , Female , Anopheles/genetics , Crosses, Genetic , DNA, Mitochondrial/genetics , DNA, Ribosomal Spacer/genetics , Anopheles/classification , Electron Transport Complex IV/genetics , Karyotyping , Republic of Korea , Sequence Analysis, DNA , ThailandABSTRACT
DNA barcoding has been widely used in species identification and biodiversity research. A short fragment of the mitochondrial cytochrome c oxidase subunit I (COI) sequence serves as a DNA bio-barcode. We collected DNA barcodes, based on COI sequences from 156 species (529 sequences) of fish, insects, and shellfish. We present results on phylogenetic relationships to assess biodiversity the in the Korean peninsula. Average GC% contents of the 68 fish species (46.9%), the 59 shellfish species (38.0%), and the 29 insect species (33.2%) are reported. Using the Kimura 2 parameter in all possible pairwise comparisons, the average interspecific distances were compared with the average intraspecific distances in fish (3.22 vs. 0.41), insects (2.06 vs. 0.25), and shellfish (3.58 vs. 0.14). Our results confirm that distance-based DNA barcoding provides sufficient information to identify and delineate fish, insect, and shellfish species by means of all possible pairwise comparisons. These results also confirm that the development of an effective molecular barcode identification system is possible. All DNA barcode sequences collected from our study will be useful for the interpretation of species-level identification and community-level patterns in fish, insects, and shellfish in Korea, although at the species level, the rate of correct identification in a diversified environment might be low.