Homo sapiens reached the higher latitudes of Europe by 45,000 years ago.

The Middle to Upper Palaeolithic transition in Europe is associated with the regional disappearance of Neanderthals and the spread of Homo sapiens. Late Neanderthals persisted in western Europe several millennia after the occurrence of H. sapiens in eastern Europe1. Local hybridization between the two groups occurred2, but not on all occasions3. Archaeological evidence also indicates the presence of several technocomplexes during this transition, complicating our understanding and the association of behavioural adaptations with specific hominin groups4. One such technocomplex for which the makers are unknown is the Lincombian-Ranisian-Jerzmanowician (LRJ), which has been described in northwestern and central Europe5-8. Here we present the morphological and proteomic taxonomic identification, mitochondrial DNA analysis and direct radiocarbon dating of human remains directly associated with an LRJ assemblage at the site Ilsenhöhle in Ranis (Germany). These human remains are among the earliest directly dated Upper Palaeolithic H. sapiens remains in Eurasia. We show that early H. sapiens associated with the LRJ were present in central and northwestern Europe long before the extinction of late Neanderthals in southwestern Europe. Our results strengthen the notion of a patchwork of distinct human populations and technocomplexes present in Europe during this transitional period.

Genetic identification of human remains from the Korean War.

The genetic identification of skeletal remains from Chinese People's Volunteers (CPVs) of the Korean War has been challenging because of the degraded DNA samples and the lack of living close relatives. This study established a workflow for identifying CPVs by combining Y-chromosome short tandem repeats (Y-STRs), mitochondrial DNA (mtDNA) hypervariable regions I and II, autosomal STRs (aSTRs), and identity-informative SNPs (iiSNPs). A total of 20 skeletal remains of CPVs and 46 samples from their alleged relatives were collected. The success rate of DNA extraction from human remains was 100%. Based on Y-STRs, six remains shared the same male lineages with their alleged relatives. Meanwhile, mtDNA genotyping supports two remains sharing the same maternal lineages with their alleged relatives. Likelihood ratios (LRs) were further obtained from 27 aSTRs and 94 iiSNPs or 1936 iiSNPs to confirm their relationship. All joint pedigree LRs were >100. Finally, six remains were successfully identified. This pilot study for the systematic genetic identification of CPVs from the Korean War can be applied for the large-scale identification of CPVs in the future.

Southern Africa's Great Escarpment as an amphitheater of climate-driven diversification and a buffer against future climate change in bats.

Hosting 1460 plant and 126 vertebrate endemic species, the Great Escarpment (hereafter, Escarpment) forms a semi-circular "amphitheater" of mountains girdling southern Africa from arid west to temperate east. Since arid and temperate biota are usually studied separately, earlier studies overlooked the biogeographical importance of the Escarpment as a whole. Bats disperse more widely than other mammalian taxa, with related species and intraspecific lineages occupying both arid and temperate highlands of the Escarpment, providing an excellent model to address this knowledge gap. We investigated patterns of speciation and micro-endemism from modeled past, present, and future distributions in six clades of southern African bats from three families (Rhinolophidae, Cistugidae, and Vespertilionidae) having different crown ages (Pleistocene to Miocene) and biome affiliations (temperate to arid). We estimated mtDNA relaxed clock dates of key divergence events across the six clades in relation both to biogeographical features and patterns of phenotypic variation in crania, bacula and echolocation calls. In horseshoe bats (Rhinolophidae), both the western and eastern "arms" of the Escarpment have facilitated dispersals from the Afrotropics into southern Africa. Pleistocene and pre-Pleistocene "species pumps" and temperate refugia explained observed patterns of speciation, intraspecific divergence and, in two cases, mtDNA introgression. The Maloti-Drakensberg is a center of micro-endemism for bats, housing three newly described or undescribed species. Vicariance across biogeographic barriers gave rise to 29 micro-endemic species and intraspecific lineages whose distributions were congruent with those identified in other phytogeographic and zoogeographic studies. Although Köppen-Geiger climate models predict a widespread replacement of current temperate ecosystems in southern Africa by tropical or arid ecosystems by 2070-2100, future climate Maxent models for 13 bat species (all but one of those analyzed above) showed minimal range changes in temperate species from the eastern Escarpment by 2070, possibly due to the buffering effect of mountains to climate change.

Mitochondrial DNA in bronchoalveolar lavage fluid is associated with the prognosis of idiopathic pulmonary fibrosis: a single cohort study.

BACKGROUND: Extracellular mitochondrial DNA (mtDNA) is released from damaged cells and increases in the serum and bronchoalveolar lavage fluid (BALF) of idiopathic pulmonary fibrosis (IPF) patients. While increased levels of serum mtDNA have been reported to be linked to disease progression and the future development of acute exacerbation (AE) of IPF (AE-IPF), the clinical significance of mtDNA in BALF (BALF-mtDNA) remains unclear. We investigated the relationships between BALF-mtDNA levels and other clinical variables and prognosis in IPF. METHODS: Extracellular mtDNA levels in BALF samples collected from IPF patients were determined using droplet-digital PCR. Levels of extracellular nucleolar DNA in BALF (BALF-nucDNA) were also determined as a marker for simple cell collapse. Patient characteristics and survival information were retrospectively reviewed. RESULTS: mtDNA levels in serum and BALF did not correlate with each other. In 27 patients with paired BALF samples obtained in a stable state and at the time of AE diagnosis, BALF-mtDNA levels were significantly increased at the time of AE. Elevated BALF-mtDNA levels were associated with inflammation or disordered pulmonary function in a stable state (n = 90), while being associated with age and BALF-neutrophils at the time of AE (n = 38). BALF-mtDNA ≥ 4234.3 copies/µL in a stable state (median survival time (MST): 42.4 vs. 79.6 months, p < 0.001) and ≥ 11,194.3 copies/µL at the time of AE (MST: 2.6 vs. 20.0 months, p = 0.03) were associated with shorter survival after BALF collection, even after adjusting for other known prognostic factors. On the other hand, BALF-nucDNA showed different trends in correlation with other clinical variables and did not show any significant association with survival time. CONCLUSIONS: Elevated BALF-mtDNA was associated with a poor prognosis in both IPF and AE-IPF. Of note, at the time of AE, it sharply distinguished survivors from non-survivors. Given the trends shown by analyses for BALF-nucDNA, the elevation of BALF-mtDNA might not simply reflect the impact of cell collapse. Further studies are required to explore the underlying mechanisms and clinical applications of BALF-mtDNA in IPF.

Unveiling the genetic structure of pig population in a Himalayan state Uttarakhand through microsatellite and mitochondrial DNA analyses.

This study traced the maternal lineage of the domestic swine populations using mitochondrial DNA control region markers and genetic diversity using microsatellite markers in Uttarakhand, an Indian state situated at the foothills of the world's youngest (geo-dynamically sensitive) mountain system, "the Himalayas". Analysis of 68 maternally unrelated individuals revealed 20 haplotypes. The maternal signature of the Pacific, Southeast Asian, European, and ubiquitously distributed Chinese haplotypes was present in Uttarakhand's domestic pig population. The D3 haplotype reported in wild pigs from North India was also identified in 47 domestic samples. A unique gene pool, UKD (Uttarakhand Domestic), as another lineage specific to this region has been proposed. Genotypes were analyzed, using 13 sets of microsatellite markers. The observed (Ho) and expected (He) heterozygosities were 0.83 ± 0.02 and 0.84 ± 0.01, respectively. The average polymorphic information content value of 0.83 ± 0.01 indicated the high informativeness of the marker. The overall mean FIS value for all the microsatellite markers was low (F = 0.04, P < 0.01). Seven loci deviated from Hardy-Weinberg equilibrium (HWE) at a significant level (p < 0.05). Two clusters were identified, indicating overlapping populations. These results suggested that though belonging to different maternal lineages, the traditional management practices in Uttarakhand have allowed for genetic mixing and the sharing of genetic material among pig populations. It could contribute to increased genetic diversity but might also result in the loss of distinct genetic characteristics or breed purity of the local breeds if not carefully managed.

Assessment of body fluid identification and DNA profiling after exposure to tropical weather conditions.

Despite current advances in body fluid identification, there are few studies evaluating the effect of environmental conditions. The present work assessed the detection of body fluids, blood, semen, and saliva, through lateral flow immunochromatographic (LFI) tests, exposed to tropical weather conditions over time, also evaluating the possibility of obtaining STR (short tandem repeat) profiles and identifying mitochondrial DNA (mtDNA) polymorphisms. Blood, semen, saliva samples, and mixtures of these fluids were deposited on polyester clothes and exposed to open-air tropical weather conditions for 1 month. The test versions from LFI (SERATEC®, Germany) Lab and crime scene (CS) used for the detection - one per each body fluid type - demonstrated that it is possible to identify body fluids and their mixtures up to 14 days after deposition. At 30 days, blood and semen were detected but not saliva. Full STR profiles were obtained from 14-day-old blood samples, and partial profiles were obtained from the remaining samples. It was possible to sequence mtDNA in the samples previously analyzed for STR profiling, and haplogroups could be assigned. In conclusion, this study demonstrated for the first time the possibility of body fluid identification and DNA profiling after exposure to tropical weather conditions for 1 month and also demonstrated the value of mtDNA analysis for compromised biological evidence.

Leveraging new methods for comprehensive characterization of mitochondrial DNA in esophageal squamous cell carcinoma.

BACKGROUND: Mitochondria play essential roles in tumorigenesis; however, little is known about the contribution of mitochondrial DNA (mtDNA) to esophageal squamous cell carcinoma (ESCC). Whole-genome sequencing (WGS) is by far the most efficient technology to fully characterize the molecular features of mtDNA; however, due to the high redundancy and heterogeneity of mtDNA in regular WGS data, methods for mtDNA analysis are far from satisfactory. METHODS: Here, we developed a likelihood-based method dMTLV to identify low-heteroplasmic mtDNA variants. In addition, we described fNUMT, which can simultaneously detect non-reference nuclear sequences of mitochondrial origin (non-ref NUMTs) and their derived artifacts. Using these new methods, we explored the contribution of mtDNA to ESCC utilizing the multi-omics data of 663 paired tumor-normal samples. RESULTS: dMTLV outperformed the existing methods in sensitivity without sacrificing specificity. The verification using Nanopore long-read sequencing data showed that fNUMT has superior specificity and more accurate breakpoint identification than the current methods. Leveraging the new method, we identified a significant association between the ESCC overall survival and the ratio of mtDNA copy number of paired tumor-normal samples, which could be potentially explained by the differential expression of genes enriched in pathways related to metabolism, DNA damage repair, and cell cycle checkpoint. Additionally, we observed that the expression of CBWD1 was downregulated by the non-ref NUMTs inserted into its intron region, which might provide precursor conditions for the tumor cells to adapt to a hypoxic environment. Moreover, we identified a strong positive relationship between the number of mtDNA truncating mutations and the contribution of signatures linked to tumorigenesis and treatment response. CONCLUSIONS: Our new frameworks promote the characterization of mtDNA features, which enables the elucidation of the landscapes and roles of mtDNA in ESCC essential for extending the current understanding of ESCC etiology. dMTLV and fNUMT are freely available from https://github.com/sunnyzxh/dMTLV and https://github.com/sunnyzxh/fNUMT , respectively.

Lepocreadiidae (Trematoda) associated with gelatinous zooplankton (Cnidaria and Ctenophora) and fishes in Australian and Japanese waters.

We examined gelatinous zooplankton from off eastern Australia for lepocreadiid trematode metacercariae. From 221 specimens of 17 species of cnidarian medusae and 218 specimens of four species of ctenophores, infections were found in seven cnidarian and two ctenophore species. Metacercariae were distinguished using cox1 mtDNA, ITS2 rDNA and morphology. We identified three species of Prodistomum Linton, 1910 [P. keyam Bray & Cribb, 1996, P. orientale (Layman, 1930), and Prodistomum Type 3], two species of Opechona Looss, 1907 [O. kahawai Bray & Cribb, 2003 and O. cf. olssoni], and Cephalolepidapedon saba Yamaguti, 1970. Two species were found in cnidarians and ctenophores, three only in cnidarians, and one only in a ctenophore. Three Australian fishes were identified as definitive hosts; four species were collected from Scomber australasicus and one each from Arripis trutta and Monodactylus argenteus. Transmission of trematodes to these fishes by ingestion of gelatinous zooplankton is plausible given their mid-water feeding habits, although such predation is rarely reported. Combined morphological and molecular analyses of adult trematodes identified two cox1 types for C. saba, three cox1 types and species of Opechona, and six cox1 types and five species of Prodistomum of which only two are identified to species. All three genera are widely distributed geographically and have unresolved taxonomic issues. Levels of distinction between the recognised species varied dramatically for morphology, the three molecular markers, and host distribution. Phylogenetic analysis of 28S rDNA data extends previous findings that species of Opechona and Prodistomum do not form monophyletic clades.

Hydrography rather than lip morphology better explains the evolutionary relationship between Gymnogeophagus labiatus and G. lacustris in Southern Brazil (Cichlidae: Geophagini)

Gymnogeophagus labiatus and G. lacustris have been long recognized as sister species exhibiting different ecological requirements. Gymnogeophagus labiatus occurs in rock bottom rivers in the hydrographic basins of Patos Lagoon (HBP) and Tramandaí River (HBT), while G. lacustris is exclusive from sand bottom coastal lagoons of the HBT. In this study, we used molecular markers, morphological measurements and data from nuptial male coloration to investigate the evolutionary relationship between these species in each hydrographic basin. We found, for all data sets, a closer relationship between G. labiatus and G. lacustris from the HBT than between G. labiatus populations from HBT and HBP. In particular, lip area had a large intraspecific plasticity, being uninformative to diagnose G. lacustris from G. labiatus. Molecular clock-based estimates suggest a recent divergence between species in the HBT (17,000 years ago), but not between G. labiatus from HBP and HBT (3.6 millions of years ago). Finally, we also found a divergent G. labiatus genetic lineage from the Camaquã River, in the HBP. These results show that the current taxonomy of G. labiatus and G. lacustris does not properly represent evolutionary lineages in these species.(AU)

Gymnogeophagus labiatus e G. lacustris vêm sendo consideradas espécies irmãs que possuem diferentes exigências ecológicas. Gymnogeophagus labiatus ocorre em rios de fundo de pedra nas bacias hidrográficas da Laguna dos Patos (HBP) e do rio Tramandaí (HBT), enquanto G. lacustris é exclusivo da HBT, ocorrendo em lagoas costeiras de fundo de arenoso. Nesse estudo, foram usados marcadores moleculares, medidas morfológicas e dados sobre a coloração nupcial em machos para investigar a relação evolutiva entre estas espécies em cada bacia hidrográfica. Para todos os conjuntos de dados foi observada uma relação mais próxima entre G. labiatus e G. lacustris da HBT do que entre as populações de G. labiatus da HBP e HBT. Em particular, a área do lábio teve uma grande plasticidade intraespecífica, não sendo informativa para diagnosticar G. lacustris de G. labiatus. Estimativas baseadas no relógio molecular sugeriram uma divergência recente entre as espécies da HBT (17.000 anos atrás), mas não entre as populações de G. labiatus da HBP e HBT (3,6 milhões de anos atrás). Finalmente, também foi encontrada uma linhagem genética de G. labiatus divergente no rio Camaquã, na HBP. Esses resultados mostram que a taxonomia atual de G. labiatus e G. lacustris não representa adequadamente as linhagens evolutivas nessas espécies.(AU)

A Century after! Rediscovery of the ancient catfish Diplomystes Bleeker 1858 (Siluriformes: Diplomystidae) in coastal river basins of Chile and its implications for conservation

The ancient catfish family Diplomystidae, with seven species endemic to rivers of southern South America, represents one of the oldest branches of the diverse order Siluriformes. With most species endangered, new reports of these species become extremely valuable for conservation. Currently, it is assumed that Diplomystes species inhabit only Andean (large) basins, and that they are extinct from coastal (small) basins from which their presence have not been recorded since 1919. Here, we document new records of the family Diplomystidae in the Laraquete and Carampangue basins, two coastal basins from the Nahuelbuta Coast Range, Chile, with no previous reports. This finding represents the rediscovery of the genus in coastal basins in more than a Century. Based on analysis of mitochondrial DNA sequences, the collected specimens were found to be closely related to Diplomystes nahuelbutaensis from the Andean Biobío Basin, but sufficiently differentiated to suggest that coastal basin populations are a different management unit. These populations are important because, contrary to previous thoughts, they prove these catfish can survive in small river networks, providing unique opportunities for research and conservation. The conservation category of Critically Endangered (CE) is recommended for the populations from the Laraquete and Carampangue basins.(AU)

La familia de bagres Diplomystidae, con siete especies endémicas de ríos del sur de Sudamérica, es uno de los linajes mas antiguos del diverso orden Siluriformes. Al estar la mayoría de las especies amenazadas, nuevos registros de éstas son extremadamente valiosos para su conservación. Actualmente, se ha asumido que los Diplomystidos se distribuyen solo en cuencas Andinas (más grandes), y que sus especies estarían extintas en cuencas de menor tamaño como las costeras, sin registros desde 1919. En este trabajo documentamos la familia Diplomistidae en las cuencas de Carampangue y Laraquete, dos cuencas costeras de la Cordillera de Nahuelbuta, Chile, lo que representa el primer registro de esta familia en estas cuencas costeras. Además, este hallazgo representa el re-descubrimiento de la familia en cuencas costeras después de un siglo. Sobre la base de análisis de ADN mitocondrial, los especímenes colectados se relacionaron más cercanamente con poblaciones de la especie Diplomystes nahuelbutaensis presente en la cuenca del Biobío. Sin embargo, existen diferencias genéticas suficientes entre las poblaciones costeras y las del Biobío para justificar su separación como unidad de manejo distinta. Estas poblaciones costeras son importantes porque demuestran que los Diplomístidos pueden sobrevivir en cuencas de pequeño tamaño, ofreciendo oportunidades únicas para su investigación y conservación. Se recomienda la categoría de conservación En Peligro Critico de Extinción (CR) para las poblaciones de las cuencas Laraquete y Carampangue.(AU)

Multiloci analyses suggest synonymy among Rhomboplites, Ocyurus and Lutjanus and reveal the phylogenetic position of Lutjanus alexandrei (Lutjanidae: Perciformes)

Lutjanidae comprises 21 genera and 135 species widespread throughout Atlantic, Indian and Pacific oceans. Nonetheless, the phylogenetic relationships of Lutjaninae remain uncertain. Furthermore, phylogenetic hypotheses for Lutjanus alexandrei, an endemic species from northeastern Brazilian coast, in Lutjanidae are absent so far. Therefore, we carried out multiloci analyses, combining both mitochondrial and nuclear DNA sequences in Lutjaninae species from Western Atlantic focusing on the controversial relationships among Lutjanus, Rhomboplites, and Ocyurus. Besides, we determined the phylogenetic position and dated the origin of L. alexandrei. The phylogenetics trees based on the 4.4 kb for 11 species corroborated the synonym among Lutjanus and the putative monotypic genera. For the dating of L. alexandrei, another nucleotide dataset (3.0 kb; 40 species) validated the genetic identity of this species that diverged from the sister taxon L. apodus between 2.5 - 6.5 Mya, probably as a result of the barrier caused by the muddy outflow from Orinoco and Amazon rivers along the coastal zone. This report is the most robust multiloci analysis to confirm the synonymy of the three genera of Lutjaninae from Western Atlantic and the first reliable inference about the phylogenetic relationships and origin of L. alexandrei.(AU)

A Família Lutjanidae compreende 21 gêneros e 135 espécies, distribuídas ao longo dos oceanos Atlântico, Índico e Pacífico. As relações filogenéticas dos Lutjaninae são incertas. Além disso, a espécie Lutjanus alexandrei, endêmica da costa nordeste do Brasil, não foi inclusa em nenhuma hipótese filogenética até o presente. Assim, realizamos uma análise integrando DNA mitocondrial e nuclear para espécies de Lutjaninae do Atlântico Ocidental, direcionada para a controversa relação entre Lutjanus, Rhomboplites e Ocyurus. Além disso, alocamos filogeneticamente L. alexandrei e datamos sua origem. As árvores filogenéticas baseadas em 4.4 kb de 11 espécies corroboraram a sinonímia entre os monotípicos e Lutjanus. Para a datação de L. alexandrei, outro banco de nuclueotídeos foi analisado (3.0 kb; 40 espécies), validando geneticamente a espécie e a colocando como irmã de L. apodus, da qual se separou entre 2.5 - 6.5 Mya, o que provavelmente foi provocado pela faixa enlameada na região costeira, influenciada pelas descargas dos rios Amazonas e Orinoco, que funciona como barreira. Este trabalho representa a mais robusta análise multiloci direcionada para a sinonimização dos três gêneros de Lutjaninae e a primeira hipótese filogenética a propor um posicionamento e origem para L. alexandrei.(AU)

Genetic variability of populations of Nyssomyia neivai in the Northern State of Paraná, Brazil

ABSTRACT The genetic study of sandfly populations needs to be further explored given the importance of these insects for public health. Were sequenced the NDH4 mitochondrial gene from populations of Nyssomyia neivai from Doutor Camargo, Lobato, Japira, and Porto Rico, municipalities in the State of Paraná, Brazil, to understand the genetic structure and gene flow. Eighty specimens of Ny. Neivai were sequenced, 20 from each municipality, and 269 base pairs were obtained. A total of 27 haplotypes and 28 polymorphic sites were found, along with a haplotypic diversity of 0.80696 and a nucleotide diversity of 0.00567. Haplotype H5, with 33 specimens, was the most common among the four populations. Only haplotypes H5 and H7 were present in all four populations. The population from Doutor Camargo showed the highest genetic diversity, and only this population shared haplotypes with those from the other municipalities. The highest number of haplotypes was sheared with Lobato which also had the highest number of unique haplotypes. This probably occurred because of constant anthropic changes that happened in the environment during the first half of the twentieth century, mainly after 1998. There was no significant correlation between genetic and geographical distances regarding these populations. However, the highest genetic and geographical distances, and the lowest gene flow were observed between Japira and Porto Rico. Geographical distance is a possible barrier between these municipalities through the blocking of haplotype sharing.

La mitocondria en la medicina legal y forense / The Mitochondrion in legal and forensic medicine

Las mitocondrias son las organelas intracelulares encargadas de suministrar la mayor parte de la energía necesaria para la actividad celular. Actúan, por lo tanto, como centrales energéticas de la célula y sintetizan ATP a expensas de los sustratos metabólicos. La intoxicación con ácido cianhídrico inhibe estos mecanismos y las alteraciones en el funcionamiento del metabolismo mitocondrial de origen genético o congénito producen innumerables patologías. El conocimiento de las patologías y disfunción de las mitocondrias es de importancia para realizar correctamente el diagnóstico de las causas de muerte. El ADN mitocondrial es de suma importancia en la medicina legal y forense para la identificación de las personas.

Mitochondria are organelles in the cell cytoplasm which supply most of the energy needed for cellular activity. They behave as cell’s power plants and synthesize ATP using metabolic substrates. Intoxication with Hydrocyanic Acid inhibits the synthesis of ATP, generating alterations in the mitochondrial metabolism, either genetic or congenital. The consequences of those alterations are innumerables pathologies. Understanding the pathologies and malfunctions of mitochondria help us to make the right diagnostic about the cause of death. In forensic medicine, mitochondrial DNA is of paramount relevance to people identification.

Molecular and morphological characterization of Contracaecum pelagicum (Nematoda) parasitizing Spheniscus magellanicus (Chordata) from Brazilian waters / Caracterização molecular e morfológica de Contracaecum pelagicum (Nematoda) parasito de Spheniscus magellanicus (Chordata) em águas brasileiras

Three new sequences of Mitochondrial cytochrome c-oxidase subunit 2 (mtDNA cox-2) from C. pelagicum parasite of Spheniscus magellanicus, the Magelanicus penguin, were determined from Brazilian waters. The sequences presented 99 and 98% of similarity with C. pelagicum sequences from Argentina, deposited on GenBank for the same genetic region and with a strong statistical support inferred from the phylogenetic tree. The morphological and ultrastructural studies that were carried out confirmed the genetic analysis.

Foram determinadas três novas sequências da região do Citocromo c-oxidase da subunidade II do DNA mitocondrial (cox-2 mtDNA) de Contracaecum pelagicum, parasito de Spheniscus magellanicus, pinguim Magalhães, de águas brasileiras. As sequências apresentaram 99 e 98% de similaridade com sequências de C. pelagicum da Argentina depositadas no GenBank da mesma região genética com forte suporte estatístico inferido pela arvore filogenética. Estudos morfológicos e ultraestruturais realizados confirmaram a identidade genética.

Especies de Anopheles presentes en el departamento del Putumayo y su infección natural con Plasmodium / Anopheles species present in the department of Putumayo and their natural infectivity with Plasmodium

Introducción. El departamento del Putumayo es una región endémica para malaria, o paludismo, causada principalmente por Plasmodium vivax . Los vectores en esta región incluyen Anopheles darlingi , el cual se ha encontrado solamente en el municipio de Puerto Leguízamo, y recientemente se incriminaron como vectores en Puerto Asís a las especies An. rangeli y An. oswaldoi . Objetivo. El propósito del trabajo fue determinar el papel de An. benarrochi B en la transmisión de malaria en este departamento, ya que se reporta como la especie más abundante que pica a los humanos. Materiales y métodos. Se recolectaron larvas y adultos de Anopheles spp. entre el 2006 y el 2008 en los municipios Puerto Leguízamo y Puerto Asís, y se obtuvieron secuencias del gen ITS-2 y del gen mitocondrial COI para confirmar las determinaciones taxonómicas por morfología. Se practicó la prueba ELISA para establecer la infección por P. vivax y P. falciparum. Resultados. Se identificaron 6.238 individuos correspondientes a 11 especies: An. albitarsis s.l. (1,83 %), An. benarrochi B (72,35 %), An. braziliensis (0,05 %), An. costai (0,06 %), An. darlingi (19,37 %), An. mattogrossensis (0,08 %), An. neomaculipalpus (0,13 %), An. oswaldoi s.l. (0,64 %), An. punctimacula (0,03 %), An. rangeli (5,12 %) y An. triannulatus s.l. (0,34 %). Se evaluaron 5.038 adultos por ELISA y 5 se encontraron positivos para P. vivax 210 y VK 247, todos pertenecientes a la especie An. benarrochi B. Conclusión. Los resultados sugieren que An. benarrochi B juega un papel en la transmisión de P. vivax en el departamento de Putumayo, dada su alta atracción por los humanos y su infección natural con Plasmodium spp.

Introduction: Putumayo is considered an endemic region for malaria transmission, mainly due to Plasmodium vivax. The vectors in this region are Anopheles darlingi , which has been found only in the municipality of Puerto Leguízamo, and An. rangeli and An. oswaldoi s.l. , which were recently incriminated as vectors in Puerto Asís. Objective: The purpose of this study was to determine the role of An. benarrochi B in malaria transmission in Putumayo, given that it is the most abundant species biting humans. Materials and methods: Collections of immature and adult stages of Anopheles spp. were made between 2006 and 2008 in the municipalities of Puerto Leguízamo and Puerto Asís in Putumayo, and sequences of internal transcribed spacer 2 ( ITS-2 ) of ribosomal DNA and the mitochondrial gene COI were obtained to confirm the morphological determinations. ELISA was carried out for P. vivax and P. falciparum infectivity. Results: A total of 6,238 specimens were identified, distributed in 11 species: An. albitarsis s.l. (1.83%), An. benarrochi B (72.35%), An. braziliensis (0.05%), An. costai (0.06%), An. darlingi (19.37%), An. mattogrossensis (0.08%), An. neomaculipalpus (0.13%), An. oswaldoi s.l. (0.64%), An. punctimacula (0.03%), An. rangeli (5.12%), and An. triannulatus s.l. (0.34%). A total of 5,038 adults were assessed by ELISA and 5 were found positive for P. vivax 210 and VK 247, all belonging to An. benarrochi B. Conclusion: The results suggest that An. benarrochi B plays a role in the transmission of P. vivax in Putumayo due to its high human contact and natural infection with Plasmodium sp.

Asociación de niveles de lípidos y haplogrupos Amerindios de DNA mitocondrial en individuos chilenos hipercolesterolémicos tratados con atorvastatina / Association of serum lipid levels and mitochondrial DNA Amerindian haplogroups in hypercholesterolemic subjects receiving atorvastatin

Resumen: Introducción: La respuesta terapéutica a estatuías se ve influenciada por factores como la edad, género y etnicidad. Con respecto a esto, el background genético de la población chilena es predominantemente Amerindio, definido por la presencia de haplogrupos Amerindios A, B, C y D de DNA mitocondrial (mtDNA). Así, el objetivo del estudio fue evaluar la potencial asociación entre la presencia de haplogrupos Amerindios de mtDNA y niveles de lípidos en individuos chilenos hipercolesterolémicos tratados con Atorvastatina. Métodos: Un total de 42 individuos en dos centros de salud del sur de Chile fueron incluidos en el estudio. En el grupo de pacientes se evaluó la presencia de haplogrupos Amerindios de mtDNA por PCR-RFLP, además de la cuantificación de Colesterol Total, Triglicéridos, Colesterol-HDL y Colesterol-LDL, antes y después del tratamiento con Atorvastatina (10 mg/día). Resultados: El 88.1 por ciento de los sujetos presentó algún haplogrupo Amerindio, no observándose diferencias en los niveles de lípidos pre- tratamiento de acuerdo al haplogrupo. Interesantemente, individuos de haplogrupo B presentaron niveles mayores de Colesterol Total (B: 254 +/- 30 mg/dl v/s C: 213 +/- 48 mg/dl, D: 230 +/- 50 mg/dl; p= 0.0319) y Colesterol-LDL (B: 157 +/- 34 mg/dl v/s C: 118 +/- 45 mg/dl, D: 135 +/- 42 mg/dl; p=0.0344) post-tratamiento. Conclusiones: El haplogrupo B se asocia a niveles mayores de lípidos post-tratamiento en pacientes tratados con Atorvastatina. Estos hallazgos sugieren por primera vez, que la presencia de haplogrupo B de mtDNA determinaría una menor respuesta al tratamiento con Atorvastatina en individuos chilenos con background genético amerindio.

Background: Therapeutic response to statins is influenced by age, gender and ethnicity. The genetic background of the Chilean population is predominantly Amerindian, defined by the presence of mitochondrial DNA (mtDNA) Amerindian haplogroups A, B, C and D Amerindian haplogroups and serum lipid levéis in hypercholesterolemic Chilean subjects receiving atorvastatin Methods: 42 subjects from southern Chile were included. The presence of mtDNA Amerindian haplogroups was evaluated by PCR-RFLP; in addition, total cholesterol, triglycerides, HDL-cholesterol and LDL-cholesterol were measured before and after treatment with atorvastatin 10 mg/day. Aim: to evaluate a possible association of mtDNA. Ameridian haplogroups and serum lipid levels in hypercholesterolemic Chilean subjects receiving atorvastatin. Result: 88.1 percent of subjects exhibited some Amerindian haplogroup. No relation of lipid levels with haplogroups was observed before treatment. Interestingly, haplogroup B individuals had higher levels of total cholesterol compared to other haplogroups after treatment (haplogroup B : 254 +/- 30 mg/dl; C : 213 +/- 48 mg/dl; D : 230 +/- 50 mg/dl, p=0.0319). Corresponding levels for LDL-cholesterol after treatment in the three groups were 157 +/- 34,118 +/-45 and 135 +/-42 mg/ di, respectively, p=0.0344. Conclusion: Compared to other haplogroups, haplogroup B is associated to higher levels of lipids after treatment with atorvastatin. For the first time, these findings suggest that the presence of mtDNA haplogroup B determines a dimished response to atorvastatin in Chilean subjets with an Amerindian genetic background.

Identificación de especies en productos de origen animal mediante PCR / PCR-species-identication in animal products

La identificación de la especie en productos de origen animal (carne, leche o sus derivados) se hace necesaria y de exigencia por los consumidores modernos, entre otras razones: i) para evitar fraude económico, ya sea por sustitución o adulteración del mismo, ii) por motivos de salud humana, tales como alergias alimentarías, iii) por implicaciones culturales; de allí que se debe contar con herramientas analíticas y sensibles para dicha identificación tales como el análisis de fragmentos de ADN en especial de origen mitocondrial (gen 12S ARNr) dada su particularidad de ser especifica de especies. A tal fin se estableció una metodología de identificación mediante la amplificación de fragmentos específicos de ADN mitocondrial (ADNm) a partir de muestras biológicas de las principales especies animales implicadas en la producción de carnes o alimentos (bovina, porcina, ovina, caprina, equina, asnos, felina y canina), específicamente de una fracción parcial del gen 12sARNr de una región conservada usando unos cebadores comunes para dichas especies, un “reverse” especifico de especie y análisis posterior mediante geles de agarosa al 1,5 por ciento y amplificación de fragmentos que oscilaron entre 150 y 364 pb. Los resultados indican que se puede identificar la especie a la que pertenece la muestra analizada en el 100 por ciento de los casos, ofreciendo una herramienta especifica para determinar la especie en alimentos de origen animal.

Nowadays identification of animal-origin products (meat, milk and dairy products) is of paramount importance to costumers and specially as for species identification for a number of reasons: i) to avoid economic fraud for substitution or alteration of the product ; ii) to avoid health issues such as food allergies; iii) for culturals reasons. There for the value of analytic and sensitive identification tools such as DNA fragments analysis, especially those of mitochondrial origin (12S rRNA gene) because of its species-specific character. An identification methodology through amplification of a species-conserved region of 12S rRNA gene (forward primer) and of a species-specific region of the same gene (reverse primer). Template DNA was extracted from biological samples of bovine, swine, ovine, caprine, equine and canine origin. After 1.5 percent agarose gel electrophoresis, fragments ranging from 150 to 364 bp were observed. Results show that species could be easily identified through PCR in all cases and that this methodology could be a specific tool for determining the origin of animal products.

Green turtles (Chelonia mydas) foraging at Arvoredo Island in Southern Brazil: genetic characterization and mixed stock analysis through mtDNA control region haplotypes

We analyzed mtDNA control region sequences of green turtles (Chelonia mydas) from Arvoredo Island, a foraging ground in southern Brazil, and identified eight haplotypes. Of these, CM-A8 (64 percent) and CM-A5 (22 percent) were dominant, the remainder presenting low frequencies (< 5 percent). Haplotype (h) and nucleotide (n) diversities were 0.5570 ± 0.0697 and 0.0021 ± 0.0016, respectively. Exact tests of differentiation and AMOVA Fi ST pairwise values between the study area and eight other Atlantic foraging grounds revealed significant differences in most areas, except Ubatuba and Rocas/Noronha, in Brazil (p > 0.05). Mixed Stock Analysis, incorporating eleven Atlantic and one Mediterranean rookery as possible sources of individuals, indicated Ascension and Aves islands as the main contributing stocks to the Arvoredo aggregation (68.01 percent and 22.96 percent, respectively). These results demonstrate the extensive relationships between Arvoredo Island and other Atlantic foraging and breeding areas. Such an understanding provides a framework for establishing adequate management and conservation strategies for this endangered species.

Populations analysis of the Brazilian Sharpnose Shark Rhizoprionodon lalandii (Chondrichthyes: Carcharhinidae) on the São Paulo coast, Southern Brazil: inferences from mt DNA sequences

Sharks of the genus Rhizoprionodon can be considered some of the most important predators along the trophic coastal marine ecosystems and represent an important economic resource for the small-scale fisheries, especially on the Brazilian coastline. In order to analyze the population structure of the shark Rhizoprionodon lalandii of São Paulo, Southeastern coast of Brazil, levels of genetic diversity were identified by nucleotide sequence analyses of the mitochondrial DNA control region. The results obtained from this study present moderate values of haplotype diversity and low nucleotide diversity. Although the AMOVA tests (ΦST = 0.08394, P < 0.01) had shown slightly differences among the studied samples, evidence for the occurrence of population structuring was not found, which may be a general feature of sharks living in coastal areas.

Tubarões do gênero Rhizoprionodon são considerados predadores de grande importância ao longo da cadeia trófica nos ecossistemas costeiros e marinhos, também representando um importante recurso econômico para a pesca, especialmente no litoral brasileiro. A fim de analisar a estrutura populacional do tubarão Rhizoprionodon lalandii no litoral de São Paulo, sudeste do Brasil, foram identificados os níveis de diversidade genética a partir da análise de sequências nucleotídicas da região controladora do DNA mitocondrial. Os dados obtidos neste estudo apresentam valores moderados de diversidade haplotípica e baixos índices de diversidade nucleotídica. Embora os testes de AMOVA (ΦST = 0,08394, P < 0,01) tenham revelado uma pequena diferença entre as amostras estudadas, evidências sobre a ocorrência de estruturação populacional não foram encontradas o que pode representar uma característica geral para tubarões vivendo em áreas costeiras.

DNA barcoding of Brazilian sea turtles (Testudines)

Five out of the seven recognized species of sea turtles (Testudines) occur on the Brazilian coast. The Barcode Initiative is an effort to undertake a molecular inventory of Earth biodiversity. Cytochrome Oxidase c subunit I (COI) molecular tags for sea turtle species have not yet been described. In this study, COI sequences for the five species of sea turtles that occur in Brazil were generated. These presented widely divergent haplotypes. All observed values were on the same range as those already described for other animal groups: the overall mean distance was 8.2 percent, the mean distance between families (Dermochelyidae and Cheloniidae) 11.7 percent, the mean intraspecific divergence 0.34 percent, and the mean distance within Cheloniidae 6.4 percent, this being 19-fold higher than the mean divergence observed within species. We obtained species-specific COI barcode tags that can be used for identifying each of the marine turtle species studied.