RESUMEN
BACKGROUND: Aedes albopictus, commonly known as the Asian tiger mosquito, has become one of the most invasive mosquito species. Over the last 5 decades, it has been introduced and established in various tropical and temperate regions worldwide. First reported in Europe in 1979 in Albania and later in Italy in 1990, the species is now established in 13 European Union (EU)/European Economic Area (EEA) countries and 337 regions (2023). In Portugal, Ae. albopictus was first detected in the Algarve and Penafiel regions in 2017, followed by Alentejo in 2022 and Lisbon in 2023. This mosquito species poses a significant public health risk as a vector for numerous pathogenic viruses, including dengue, Zika, and chikungunya. METHODS: Aedes albopictus collected in Lisbon in 2023 were analyzed using cytochrome c oxidase I (COX) gene sequencing to understand their genetic relationships. RESULTS: Our data indicate that the Ae. albopictus mosquito populations detected in three locations in Lisbon in 2023 correspond to recent but distinct introduction events. CONCLUSIONS: Although there has been no local transmission of Aedes-transmitted viruses in mainland Portugal to date, the spread of the mosquito and increased international travel increase the risk of Aedes-borne disease outbreaks. The ongoing spread of Ae. albopictus in the country and the confirmed multiple introductions in new locations raise awareness of the need to monitor mosquito vectors to control and prevent autochthonous Aedes-borne disease outbreaks.
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Aedes , Especies Introducidas , Mosquitos Vectores , Aedes/genética , Aedes/virología , Aedes/clasificación , Animales , Portugal , Mosquitos Vectores/genética , Mosquitos Vectores/virología , Filogenia , Complejo IV de Transporte de Electrones/genética , FemeninoRESUMEN
BACKGROUND: DNA metabarcoding applies high-throughput sequencing approaches to generate numerous DNA barcodes from mixed sample pools for mass species identification and community characterisation. To date, however, most metabarcoding studies employ second-generation sequencing platforms like Illumina, which are limited by short read lengths and longer turnaround times. While third-generation platforms such as the MinION (Oxford Nanopore Technologies) can sequence longer reads and even in real-time, application of these platforms for metabarcoding has remained limited possibly due to the relatively high read error rates as well as the paucity of specialised software for processing such reads. RESULTS: We show that this is no longer the case by performing nanopore-based, cytochrome c oxidase subunit I (COI) metabarcoding on 34 zooplankton bulk samples, and benchmarking the results against conventional Illumina MiSeq sequencing. Nanopore R10.3 sequencing chemistry and super accurate (SUP) basecalling model reduced raw read error rates to ~ 4%, and consensus calling with amplicon_sorter (without further error correction) generated metabarcodes that were ≤ 1% erroneous. Although Illumina recovered a higher number of molecular operational taxonomic units (MOTUs) than nanopore sequencing (589 vs. 471), we found no significant differences in the zooplankton communities inferred between the sequencing platforms. Importantly, 406 of 444 (91.4%) shared MOTUs between Illumina and nanopore were also found to be free of indel errors, and 85% of the zooplankton richness could be recovered after just 12-15 h of sequencing. CONCLUSION: Our results demonstrate that nanopore sequencing can generate metabarcodes with Illumina-like accuracy, and we are the first study to show that nanopore metabarcodes are almost always indel-free. We also show that nanopore metabarcoding is viable for characterising species-rich communities rapidly, and that the same ecological conclusions can be obtained regardless of the sequencing platform used. Collectively, our study inspires confidence in nanopore sequencing and paves the way for greater utilisation of nanopore technology in various metabarcoding applications.
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Código de Barras del ADN Taxonómico , Secuenciación de Nucleótidos de Alto Rendimiento , Nanoporos , Código de Barras del ADN Taxonómico/métodos , Animales , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Mutación INDEL , Secuenciación de Nanoporos/métodos , Complejo IV de Transporte de Electrones/genética , Zooplancton/genética , Zooplancton/clasificación , Análisis de Secuencia de ADN/métodosRESUMEN
BACKGROUND: Western Ghats is a biodiversity treasure trove with reports of indigenous leishmaniasis cases. Hence, systematic sand fly surveillance was carried out among the tribal population. The present study reports a novel sand fly species, Phlebotomus (Anaphlebotomus) ajithii n. sp. (Diptera: Psychodidae), discovered in the Western Ghats of India. METHODS: A comprehensive sand fly survey was conducted across the Kollam, Thrissur, Idukki, Kasaragod and Malappuram districts of Kerala, India. The survey spanned both indoor and outdoor habitats using standard collection methods over a 3-year, 3-month period. DNA barcoding of samples was performed targeting mitochondrial cytochrome c oxidase subunit I (COI) gene, and the sequence generated was subjected to phylogenetic analysis. RESULTS: Phlebotomus (Anaphlebotomus) ajithii, a new sand fly species, is recorded and described in this communication. The morphological relationship of the new species to other members of the subgenus Anaphlebotomus is discussed. Mitochondrial COI barcode followed by phylogenetic analysis confirmed that specimens of Ph. ajithii belong to the same taxonomic group, while a genetic distance of 11.7% from congeners established it as a distinct species. CONCLUSIONS: The Western Ghats, known for its rich biodiversity, has lacked systematic entomological surveys focusing on sand flies. This study aims to fill this gap and reports and describes a new species of sand fly.
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Complejo IV de Transporte de Electrones , Phlebotomus , Filogenia , Animales , India , Phlebotomus/genética , Phlebotomus/clasificación , Complejo IV de Transporte de Electrones/genética , Código de Barras del ADN Taxonómico , Femenino , MasculinoRESUMEN
BACKGROUND: Black flies are among the most medically and veterinary important insects, as adult females of certain species are the sole vector of Onchocerca volvulus. Here, a new black fly species belonging to the subgenus Asiosimulium Takaoka & Choochote, 2005, is described and formally named as Simulium (Asiosimulium) kittipati sp. nov. METHODS: Pupae and larvae of black flies were collected from available substrates in the stream from central Thailand. Pupae were individually separated in plastic tubes and maintained until adult flies emerged. The emerged adult flies associated with their pupal exuviae and cocoon as well as mature larvae preserved in 85% ethanol were used to describe the new species based on an integrated approach of morphological examination and molecular analysis of the COI gene. RESULTS: The new species is characterized in the female by the medium-long sensory vesicle with a medium-sized opening apically, scutum with three faint longitudinal vittae, and the ellipsoidal spermatheca; in the male by the number of upper-eye (large) facets in 20 vertical columns and 21 horizontal rows, hind basitarsus slender, nearly parallel-sided, and median sclerite much wider and upturned apically; in the pupa by the head and thoracic integument densely covered with tiny tubercles, and the pupal gill of arborescent type with 28-30 filaments; and in the larva by the postgenal cleft deep, nearly reaching the posterior margin of the hypostoma, and dark pigmented sheath of the subesophageal ganglion. The DNA barcode successfully differentiated the new species from its congeners with an interspecific genetic divergence of 1.74-18.72%, confirming the morphological identification that the species is a new member of the subgenus Asiosimulium. Phylogenetic analyses also indicated that the new species is genetically closely related to Simulium phurueaense Tangkawanit, Wongpakam & Pramual, 2018, further supporting its morphological classification. CONCLUSIONS: This is the ninth species assigned to the subgenus Asiosimulium within the genus Simulium Latreille, 1802. Taxonomic notes and identification keys are given to distinguish this new species from the eight known species members in its same subgenus. Additionally, a distribution map of all species members in this subgenus occurring in Thailand and other countries is provided.
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Larva , Filogenia , Pupa , Simuliidae , Animales , Simuliidae/genética , Simuliidae/anatomía & histología , Simuliidae/clasificación , Tailandia , Femenino , Masculino , Pupa/anatomía & histología , Pupa/genética , Pupa/clasificación , Larva/anatomía & histología , Larva/genética , Larva/clasificación , Complejo IV de Transporte de Electrones/genética , Insectos Vectores/anatomía & histología , Insectos Vectores/genética , Insectos Vectores/clasificaciónRESUMEN
Background: Non-steroidal anti-inflammatory drugs (NSAIDs), such as diclofenac (DCF), form a significant group of environmental contaminants. When the toxic effects of DCF on plants are analyzed, authors often focus on photosynthesis, while mitochondrial respiration is usually overlooked. Therefore, an in vivo investigation of plant mitochondria functioning under DCF treatment is needed. In the present work, we decided to use the green alga Chlamydomonas reinhardtii as a model organism. Methods: Synchronous cultures of Chlamydomonas reinhardtii strain CC-1690 were treated with DCF at a concentration of 135.5 mg × L-1, corresponding to the toxicological value EC50/24. To assess the effects of short-term exposure to DCF on mitochondrial activity, oxygen consumption rate, mitochondrial membrane potential (MMP) and mitochondrial reactive oxygen species (mtROS) production were analyzed. To inhibit cytochrome c oxidase or alternative oxidase activity, potassium cyanide (KCN) or salicylhydroxamic acid (SHAM) were used, respectively. Moreover, the cell's structure organization was analyzed using confocal microscopy and transmission electron microscopy. Results: The results indicate that short-term exposure to DCF leads to an increase in oxygen consumption rate, accompanied by low MMP and reduced mtROS production by the cells in the treated populations as compared to control ones. These observations suggest an uncoupling of oxidative phosphorylation due to the disruption of mitochondrial membranes, which is consistent with the malformations in mitochondrial structures observed in electron micrographs, such as elongation, irregular forms, and degraded cristae, potentially indicating mitochondrial swelling or hyper-fission. The assumption about non-specific DCF action is further supported by comparing mitochondrial parameters in DCF-treated cells to the same parameters in cells treated with selective respiratory inhibitors: no similarities were found between the experimental variants. Conclusions: The results obtained in this work suggest that DCF strongly affects cells that experience mild metabolic or developmental disorders, not revealed under control conditions, while more vital cells are affected only slightly, as it was already indicated in literature. In the cells suffering from DCF treatment, the drug influence on mitochondria functioning in a non-specific way, destroying the structure of mitochondrial membranes. This primary effect probably led to the mitochondrial inner membrane permeability transition and the uncoupling of oxidative phosphorylation. It can be assumed that mitochondrial dysfunction is an important factor in DCF phytotoxicity. Because studies of the effects of NSAIDs on the functioning of plant mitochondria are relatively scarce, the present work is an important contribution to the elucidation of the mechanism of NSAID toxicity toward non-target plant organisms.
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Antiinflamatorios no Esteroideos , Chlamydomonas reinhardtii , Diclofenaco , Potencial de la Membrana Mitocondrial , Mitocondrias , Consumo de Oxígeno , Especies Reactivas de Oxígeno , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Mitocondrias/patología , Diclofenaco/toxicidad , Chlamydomonas reinhardtii/efectos de los fármacos , Chlamydomonas reinhardtii/metabolismo , Chlamydomonas reinhardtii/ultraestructura , Antiinflamatorios no Esteroideos/toxicidad , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Consumo de Oxígeno/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Complejo IV de Transporte de Electrones/metabolismo , Cianuro de Potasio/toxicidad , Oxidorreductasas/metabolismo , Salicilamidas , Microscopía Electrónica de Transmisión , Proteínas de Plantas , Proteínas MitocondrialesRESUMEN
Background: Ariidae species play a significant role as fishing resources in the Amazon region. However, the family's systematic classification is notably challenging, particularly regarding species delimitation within certain genera. This difficulty arises from pronounced morphological similarities among species, posing obstacles to accurate species recognition. Methods: Following morphological identification, mitochondrial markers (COI and Cytb) were employed to assess the diversity of Ariidae species in the Amazon. Results: Our sampling efforts yielded 12 species, representing 92% of the coastal Amazon region's diversity. Morphological identification findings were largely corroborated by molecular data, particularly for species within the Sciades and Bagre genera. Nonetheless, despite morphological support, Cathorops agassizii and Cathorops spixii displayed minimal genetic divergence (0.010). Similarly, Notarius quadriscutis and Notarius phrygiatus formed a single clade with no genetic divergence, indicating mitochondrial introgression. For the majority of taxa examined, both COI and Cytb demonstrated efficacy as DNA barcodes, with Cytb exhibiting greater polymorphism and resolution. Consequently, the molecular tools utilized proved highly effective for species discrimination and identification.
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Bagres , Código de Barras del ADN Taxonómico , ADN Mitocondrial , Animales , Bagres/genética , Bagres/clasificación , ADN Mitocondrial/genética , Filogenia , Variación Genética/genética , Brasil , Complejo IV de Transporte de Electrones/genéticaRESUMEN
Requiem sharks (genus Carcharhinus) have previously been reported to form large aggregations around marine infrastructures in the eastern Mediterranean Sea. While this behaviour may offer fitness advantages at the individual level, the implications of extended residency at human-altered habitats for population persistence have yet to be assessed. In this work, we investigated the phylogeographic and demographic composition of sharks near a coal-fired power and desalination station in Israel. Our aim was to infer habitat use and the mechanisms underlying the aggregation behaviour, and to highlight potential conservation impacts. We sampled, measured, and released 70 individuals between 2016 and 2022 to assess genetic variability within the cytochrome C oxidase I (COI) region and to analyse the aggregation's structure based on the sharks' size and sex distribution. In addition, we performed meristic counts on a reference specimen collected dead at another power station in Israel to supplement species identification using the abovementioned techniques. Our findings indicate size-based sex segregation of adult female dusky and male sandbar sharks (Carcharhinus obscurus and Carcharhinus plumbeus, respectively), with each species comprising two COI haplotypes. In the dusky shark, one haplotype corresponded to an Indo-Pacific lineage, and the other matched an Atlantic lineage. In the sandbar shark, we observed a haplotype previously sampled in the Mediterranean Sea, the Red Sea, the Northwest Indian Ocean, and South Africa, and another haplotype that was unique to our study site and genetically closer to the former than to sequences sampled in other ocean basins. This study provides the first indication of sympatric aggregation amongst phylogeographically distinct dusky and sandbar shark lineages, suggesting that human-altered habitats in the eastern Mediterranean Sea may influence the distribution of these species. Based on the observed segregation pattern, we conclude that the site does not function as a nursery, parturition, or mating area, and discuss other plausible explanations that warrant further research. Finally, we highlight important directions for future research and the implications of our findings for management and conservation.
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Ecosistema , Complejo IV de Transporte de Electrones , Haplotipos , Tiburones , Tiburones/genética , Tiburones/clasificación , Mar Mediterráneo , Animales , Femenino , Masculino , Complejo IV de Transporte de Electrones/genética , Humanos , Israel , Variación Genética , Filogenia , FilogeografíaRESUMEN
Ichthyoplankton monitoring is crucial for stock assessments, offering insights into spawning grounds, stock size, seasons, recruitment, and changes in regional ichthyofauna. This study evaluates the efficiency of multi-marker DNA metabarcoding using mitochondrial cytochrome c oxidase subunit I (COI), 12S rRNA and 16S rRNA gene markers, in comparison to morphology-based methods for fish species identification in ichthyoplankton samples. Two transects with four coastal distance categories were sampled along the southern coast of Portugal, being each sample divided for molecular and morphological analyses. A total of 76 fish species were identified by both approaches, with DNA metabarcoding overperforming morphology-75 versus 11 species-level identifications. Linking species-level DNA identifications with higher taxonomic morphological identifications resolved several uncertainties associated with traditional methods. Multi-marker DNA metabarcoding improved fish species detection by 20-36% compared to using a single marker/amplicon, and identified 38 species in common, reinforcing the validity of our results. PERMANOVA analysis revealed significant differences in species communities based on the primer set employed, transect location, and distance from the coast. Our findings underscore the potential of DNA metabarcoding to assess ichthyoplankton diversity and suggest that its integration into routine surveys could enhance the accuracy and comprehensiveness of fish stock assessments.
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Código de Barras del ADN Taxonómico , Peces , ARN Ribosómico 16S , Animales , Código de Barras del ADN Taxonómico/métodos , Peces/genética , Peces/clasificación , ARN Ribosómico 16S/genética , Complejo IV de Transporte de Electrones/genética , Marcadores Genéticos , Portugal , ARN Ribosómico/genética , Biodiversidad , Zooplancton/genética , Zooplancton/clasificaciónRESUMEN
BACKGROUND: Opisthorchis-like eggs are a public health problem in northern and northeastern Thailand. However, the genetic epidemiology and structure of these parasites in northern Thailand are unknown. Thus, this study investigated their population genetic structure using cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) nucleotide sequences. METHODOLOGY/PRINCIPAL FINDINGS: A study was conducted in the hill tribe regions of Chiang Mai Province, northern Thailand. Internal transcribed spacer 2 polymerase chain reaction and restriction fragment length polymorphism were used to distinguish 205 positive feces samples for Opisthorchis-like eggs. The results showed that the prevalence of O. viverrini and Haplorchis taichui was 10.5% and 38.2%, respectively, and the co-infection rate was 37.2%. To determine the genetic structure of O. viverrini and H. taichui using cox1 and nad1 genes, genetic analysis was performed using 30 randomly chosen fecal samples for Opisthorchis-like eggs. Pairwise FST analysis indicated that O. viverrini and H. taichui displayed nonsignificant genetic differentiation within Chiang Mai Province and between interpopulations from different geographic areas. Moreover, within the intrapopulation in Chiang Mai Province, cox1 presented higher gene flow than nad1 in O. viverrini, while nad1 demonstrated higher gene flow than cox1 in H. taichui. The neutrality tests based on Fu's Fs indicated population expansion and selective sweep from bottleneck or hitchhiking in O. viverrini and H. taichui populations, supported by haplotype network patterns. Phylogenetic tree analysis based on cox1 and nad1 revealed the monophyly of O. viverrini and H. taichui and genetic relationships with other isolates collected from Thailand, Lao People's Democratic Republic (PDR), and Vietnam. CONCLUSIONS/SIGNIFICANCE: This study investigated the molecular discrimination and genetic structure of Opisthorchis-like eggs in northern Thailand. The genetic information derived from this study could be associated with the background, molecular epidemiology, and disease severity of these parasites.
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Complejo IV de Transporte de Electrones , Heces , Opistorquiasis , Opisthorchis , Animales , Tailandia/epidemiología , Opistorquiasis/parasitología , Opistorquiasis/epidemiología , Opisthorchis/genética , Opisthorchis/clasificación , Opisthorchis/aislamiento & purificación , Heces/parasitología , Humanos , Complejo IV de Transporte de Electrones/genética , NADH Deshidrogenasa/genética , Genes Mitocondriales/genética , Variación Genética , Polimorfismo de Longitud del Fragmento de Restricción , Filogenia , PrevalenciaRESUMEN
OBJECTIVES: Cox10 is an enzyme required for the activity of cytochrome c oxidase. Humans who lack at least one functional copy of Cox10 have a form of Leigh Syndrome, a genetic disease that is usually fatal in infancy. As more human genomes are sequenced, new alleles are being discovered; whether or not these alleles encode functional proteins remains unclear. Thus, we set out to measure the phenotypes of many human Cox10 variants by expressing them in yeast cells. RESULTS: We successfully expressed the reference sequence and 25 variants of human Cox10 in yeast. We quantitated the ability of these variants to support growth on nonfermentable media and directly measured cytochrome c oxidase activity. 11 of these Cox10 variants supported approximately half or more the cytochrome c oxidase activity compared to the reference sequence. All of the strains containing those 11 variants also grew robustly using a nonfermentable carbon source. Cells expressing the other variants showed low cytochrome c oxidase activity and failed to grow on nonfermentable media.
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Complejo IV de Transporte de Electrones , Enfermedad de Leigh , Fenotipo , Enfermedad de Leigh/genética , Complejo IV de Transporte de Electrones/genética , Complejo IV de Transporte de Electrones/metabolismo , Humanos , Saccharomyces cerevisiae/genética , Variación Genética , Proteínas de la Membrana , Transferasas Alquil y ArilRESUMEN
BACKGROUND: The larvae of Altenia spp. and gall aphids are known to feed on plants related to Anacardiaceae. This study documents the aphidophagous habit of Altenia wagneriella, which was verified by molecular techniques, subsequently by the gut dissection test, and direct observation. MATERIALS AND METHODS: To identify the moth larvae and adult aphids, two mitochondrial genes, cytochrome c oxidase I (COI) and cytochrome b (Cytb), were amplified by polymerase chain reaction (PCR). Nested PCR with the aphid-specific primer pairs AphidF and AphidR was used to detect aphids in the body of moth larvae. The specificity of the primers was verified by PCR analysis of DNA from moth larvae and adult aphids. RESULTS: The method detected aphids in moth larvae, and a band of approximately 200 bp was amplified from moth larvae feeding on aphids. No cross reactions with moth larvae were observed. In the laboratory, all moth larvae feeding on aphids (Forda marginata) were also PCR positive for aphids. CONCLUSIONS: Gall-inducing insects are microhabitat engineers that manipulate their host to obtain a better nutrient supply, as well as protection from natural enemies and abiotic factors. This is the first recorded instance worldwide of the carnivorous larva of the moth A. wagneriella acting as an aphid predator, as well as the first record of a host insect for this species. Additionally, it is the first effort to molecularly analyze the predator-prey relationship between the moth larvae and the aphids inside the wild pistachio gall.
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Áfidos , Larva , Mariposas Nocturnas , Conducta Predatoria , Animales , Áfidos/genética , Áfidos/fisiología , Irán , Larva/genética , Mariposas Nocturnas/genética , Complejo IV de Transporte de Electrones/genética , Citocromos b/genética , Reacción en Cadena de la Polimerasa/métodosRESUMEN
Oxidative Phosphorylation (OXPHOS) defects can cause severe encephalopathies and no effective treatment exists for these disorders. To assess the ability of gene replacement to prevent disease progression, we subjected two different CNS-deficient mouse models (Ndufs3/complex I or Cox10/complex IV conditional knockouts) to gene therapy. We used retro-orbitally injected AAV-PHP.eB to deliver the missing gene to the CNS of these mice. In both cases, we observed survival extension from 5-6 to more than 15 months, with no detectable disease phenotypes. Likewise, molecular and cellular phenotypes were mostly recovered in the treated mice. Surprisingly, these remarkable phenotypic improvements were achieved with only ~30% of neurons expressing the transgene from the AAV-PHP.eB vector in the conditions used. These findings suggest that neurons lacking OXPHOS are protected by the surrounding neuronal environment and that partial compensation for neuronal OXPHOS loss can have disproportionately positive effects.
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Modelos Animales de Enfermedad , Ratones Noqueados , Encefalomiopatías Mitocondriales , Neuronas , Fosforilación Oxidativa , Animales , Neuronas/metabolismo , Ratones , Encefalomiopatías Mitocondriales/metabolismo , Encefalomiopatías Mitocondriales/genética , Encefalomiopatías Mitocondriales/terapia , Terapia Genética/métodos , Complejo I de Transporte de Electrón/metabolismo , Complejo I de Transporte de Electrón/genética , Complejo I de Transporte de Electrón/deficiencia , Complejo IV de Transporte de Electrones/metabolismo , Vectores Genéticos/metabolismo , Dependovirus/genética , Proteínas de la Membrana , Transferasas Alquil y ArilRESUMEN
In this study, a tick intracellular symbiont, Candidatus Midichloria mitochondrii, was detected in Hyalomma anatolicum from Xinjiang, China. Morphological identification and cytochrome oxidase subunit I sequence alignment were used for molecular identification of the tick species. PCR detection further revealed the presence of endosymbiont C. M. mitochondrii in the tick. Specific primers were designed for Groel and 16S rRNA genes of C. M. mitochondrii for PCR amplification and phylogenetic analysis. To further investigate the vertical transmission characteristics of C. M. mitochondrii, specific primers were designed based on the Fabâ gene fragment to detect C. M. mitochondrii in different developmental stages and organs of the tick using qPCR. Of the 336 tick specimens collected from the field, 266 samples were identified as H. anatolicum on the basis of morphological characteristics. The gene fragment alignment results of COI confirmed that these ticks were H. anatolicum. The phylogenetic analysis showed that Groel gene of C. M. mitochondrii clustered with Midichloria strains detected in Ixodes ricinus ticks from Italy and Ixodes holocyclus ticks from Australia, with 100% sequence similarity. Furthermore, the 16S rRNA gene of C. M. mitochondrii clusters with the strains isolated from Hyalomma rufipes ticks in Italy, exhibiting the highest degree of homology. qPCR results showed that C. M. mitochondrii was present at all developmental stages of H. anatolicum, with the highest relative abundance in eggs, and lower relative abundance in nymphs and unfed males. With female tick blood feeding, the relative abundance of C. M. mitochondrii increased, and a particularly high relative abundance was detected in the ovaries of engorged female ticks. This study provides information for studying the survival adaptability of H. anatolicum, and provides data for further investigation of the mechanisms regulating tick endosymbionts in ticks, enriching the reference materials for comprehensive prevention and control of tick-borne diseases.
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Ixodidae , Filogenia , ARN Ribosómico 16S , Simbiosis , Animales , Ixodidae/microbiología , Ixodidae/crecimiento & desarrollo , ARN Ribosómico 16S/genética , Femenino , Masculino , China , Chaperonina 60/genética , Ninfa/microbiología , Ninfa/crecimiento & desarrollo , Alineación de Secuencia , Complejo IV de Transporte de Electrones/genética , Infestaciones por Garrapatas/parasitología , Infestaciones por Garrapatas/veterinaria , Reacción en Cadena de la Polimerasa , ADN Bacteriano , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
Three species of Hemiscorpius were identified in Hormozgan province, for which the available antivenoms lack the efficacy required for treating patients. Consequently, an exact identification of the existing species was deemed necessary as the first step in managing treatment procedures. Considering the morphological similarities among the species, the aim of this research was the molecular study of the samples to accurately identify the species. Hemiscorpius specimens were collected from various locations in Hormozgan province between 2021 to 2023. The Cytochrome c oxidase subunit I gene was amplified and sequenced. Four sequences were obtained from Hemiscorpius specimens collected from Hormozgan province, and three sequences were sourced from the NCBI for analysis. Bayesian inference and Maximum likelihood phylogenetic trees showed similar results, positioning the base of Hemiscorpius enischnochela tree as an older species and Hemiscorpius lepturus adjacent to Hemiscorpius acanthocercus, identified as the newest species at the tree's tip. The results confirmed the validity of three species, namely H. acanthocercus, H. enischnochela, and H. lepturus. Hemiscorpius acanthocercus and H. lepturus are known for having dangerous venom for humans with reported deaths due to their stings. Considering the importance of the members of this genus from the medical point of view, a comprehensive examination of all species is imperative.
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Complejo IV de Transporte de Electrones , Filogenia , Escorpiones , Animales , Irán , Complejo IV de Transporte de Electrones/análisis , Complejo IV de Transporte de Electrones/genética , Escorpiones/genética , Escorpiones/clasificación , Animales PonzoñososRESUMEN
Cytochrome c oxidase (Cox) is a crucial terminal oxidase in the electron transport chain. In this study, we generated 14 Cox gene deletion or overexpression mutants in Fusarium graminearum. Fungicide sensitivity tests revealed that 11 Cox gene deletion mutants displayed resistance to pyraclostrobin, while 10 overexpression mutants showed hypersensitivity. RNA-Seq and RT-qPCR analyses demonstrated the upregulation of FgAox (alternative oxidase in F. graminearum), FgAod2, and FgAod5 (alternative oxidase deficiency in F. graminearum) in ΔFgCox4-2 and ΔFgCox17-75 mutants. In 11 Cox gene deletion mutants, FgAox expression was significantly upregulated, whereas in 10 Cox gene overexpression mutants, it was significantly downregulated. FgAox overexpression mutants exhibit resistance to pyraclostrobin, while FgAox deletion mutants show hypersensitivity to pyraclostrobin. FgAod2 and FgAod5 were identified as transcription factors for FgAox. Our findings reveal that FgCox influences pyraclostrobin sensitivity by regulating FgAox through FgAod2 and FgAod5. Understanding pyraclostrobin resistance mechanisms in F. graminearum could help develop better fungicide rotation and application strategies to manage resistance and guide the creation of new fungicides targeting different pathways.
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Complejo IV de Transporte de Electrones , Proteínas Fúngicas , Fungicidas Industriales , Fusarium , Estrobilurinas , Factores de Transcripción , Fusarium/genética , Fusarium/enzimología , Estrobilurinas/farmacología , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Fungicidas Industriales/farmacología , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Complejo IV de Transporte de Electrones/genética , Complejo IV de Transporte de Electrones/metabolismo , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/genética , Regulación Fúngica de la Expresión Génica , Oxidorreductasas/genética , Oxidorreductasas/metabolismoRESUMEN
The mitochondrial cytochrome c oxidase subunit I (COI) genes of six endangered goose breeds (Xupu, Yangjiang, Yan, Wuzong, Baizi, and Lingxian) were sequenced and compared to assess the genetic diversity of endangered goose breeds. By constructing phylogenetic trees and evolutionary maps of genetic relationships, the affinities and degrees of genetic variations among the six different breeds were revealed. A total of 92 polymorphic sites were detected in the 741 bp sequence of the mtDNA COI gene after shear correction, and the GC content of the processed sequence (51.11%) was higher than that of the AT content (48.89%). The polymorphic loci within the populations of five of the six breeds (Xupu, Yangjiang, Yan, Baizi, and Lingxian) were more than 10, the haplotype diversity > 0.5, and the nucleotide diversity (Pi) > 0.005, with the Baizi geese being the exception. A total of 35 haplotypes were detected based on nucleotide variation among sequences, and the goose breed haplotypes showed a central star-shaped dispersion; the FST values were -0.03781 to 0.02645, The greatest genetic differentiation (FST = 0.02645) was observed in Yan and Wuzong breeds. The most frequent genetic exchange (Nm > 15.00) was between the Wuzong and Yangjiang geese. An analysis of molecular variance showed that the population genetic variation mainly came from within the population; the base mismatch differential distribution analysis of the goose breeds and the Tajima's D and Fu's Fs neutral detection of the historical occurrence dynamics of their populations were negative (p > 0.10). The distribution curve of the base mismatches showed a multimodal peak, which indicated that the population tended to be stabilised. These results provide important genetic information for the conservation and management of endangered goose breeds and a scientific basis for the development of effective conservation strategies.
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Complejo IV de Transporte de Electrones , Especies en Peligro de Extinción , Gansos , Haplotipos , Filogenia , Animales , Gansos/genética , Complejo IV de Transporte de Electrones/genética , Variación Genética , ADN Mitocondrial/genética , Cruzamiento , China , Mitocondrias/genéticaRESUMEN
Forensic entomology plays a crucial role in criminal investigations by providing vital insights into minimum postmortem interval (PMImin) and corpse relocation by identifying insect species that colonize in decomposing remains. This study aimed to identify and analyze the genetic variation of forensically significant fly species in Thailand, using DNA barcoding of the mitochondrial cytochrome c oxidase subunit I COI gene. A total of 3,220 fly specimens were collected from 18 provinces across six regions of Thailand from October 2017 to September 2022. These specimens were classified by morphological identification into 21 species among three Dipteran families: Calliphoridae, Muscidae, and Sarcophagidae, with Chrysomya megacephala Diptera: Calliphoridae being the most abundant species. DNA barcoding confirmed the morphological identifications with 100 % accuracy, showing low intraspecific K2P distances0.0 to 1.1 %) and significant interspecific K2P distances 2.5 % to 17.2 %. A Neighbour-Joining (NJ) analysis was conducted to assess the molecular identification capabilities of the barcoding region. This analysis successfully recovered nearly all species as distinct monophyletic groups. The species groupings obtained were generally consistent with both morphological and molecular identifications. These findings underscore the effectiveness of DNA barcoding for precise species identification and contribute to a comprehensive database of forensically important flies in Thailand, thus facilitating improved forensic investigations and biodiversity studies.
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Código de Barras del ADN Taxonómico , Complejo IV de Transporte de Electrones , Entomología Forense , Variación Genética , Animales , Tailandia , Complejo IV de Transporte de Electrones/genética , Dípteros/genética , Dípteros/clasificación , Dípteros/anatomía & histología , Calliphoridae/genética , Calliphoridae/clasificación , Filogenia , Sarcofágidos/genética , Sarcofágidos/clasificación , Muscidae/genética , Muscidae/clasificaciónRESUMEN
Cytochrome c oxidase (CcO) reduces O2, pumps protons in the mitochondrial respiratory chain, and is essential for oxygen consumption in the cell. The coiled-coil-helix-coiled-coil-helix domain-containing 2 (CHCHD2; also known as mitochondrial nuclear retrograde regulator 1 [MNRR1], Parkinson's disease 22 [PARK22] and aging-associated gene 10 protein [AAG10]) is a protein that binds to CcO from the intermembrane space and positively regulates the activity of CcO. Despite the importance of CHCHD2 in mitochondrial function, the mechanism of action of CHCHD2 and structural information regarding its binding to CcO remain unknown. Here, we utilized visible resonance Raman spectroscopy to investigate the structural changes around the hemes in CcO in the reduced and CO-bound states upon CHCHD2 binding. We found that CHCHD2 has a significant impact on the structure of CcO in the reduced state. Mapping of the heme peripheries that result in Raman spectral changes in the structure of CcO highlighted helices IX and X near the hemes as sites where CHCHD2 takes action. Part of helix IX is exposed in the intermembrane space, whereas helix X, located between both hemes, may play a key role in proton uptake to a proton-loading site in the reduced state for proton pumping. Taken together, our results suggested that CHCHD2 binds near helix IX and induces a structural change in helix X, accelerating proton uptake.
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Proteínas de Unión al ADN , Complejo IV de Transporte de Electrones , Hemo , Proteínas Mitocondriales , Espectrometría Raman , Factores de Transcripción , Espectrometría Raman/métodos , Complejo IV de Transporte de Electrones/química , Complejo IV de Transporte de Electrones/metabolismo , Hemo/química , Hemo/metabolismo , Proteínas de Unión al ADN/química , Proteínas de Unión al ADN/metabolismo , Proteínas Mitocondriales/química , Proteínas Mitocondriales/metabolismo , Factores de Transcripción/metabolismo , Factores de Transcripción/química , Humanos , Unión ProteicaRESUMEN
Phlebotomine sand flies are recognized as a primary vector of Leishmania and are also suspected vectors of Trypanosoma. The transmission cycle of these parasites relies on the distribution of sand fly vectors, parasites, and reservoir animals. This study aimed to detect Leishmania and Trypanosoma DNA and identify the sources of bloodmeals in post-feeding sand flies captured across Thailand. A total of 42,911 field female sand flies were collected from 11 provinces across Thailand using CDC light traps. Among these, 253 post-feeding sand flies were selected for analysis. The predominant species in this study was Sergentomyia khawi (33.60 %). The DNA was extracted from individual female sand flies. Polymerase chain reaction (PCR), specific to the internal transcribed spacer 1 (ITS1) and the small subunit ribosomal RNA (SSU rRNA) gene regions were used to detect the presence of Leishmania and Trypanosoma DNA, respectively. Additionally, cytochrome c oxidase subunit I (COI) gene region was utilized to identify the sources of host bloodmeals. Leishmania DNA was not detected in any specimens. The analysis of SSU rRNA sequences revealed the presence of Trypanosoma DNA (11.46 %, 29/253) in sand fly samples. Among these samples, T. noyesi (1.58 %, 4/253) was identified in Idiophlebotomus longiforceps and Phlebotomus asperulus, Trypanosoma Anura01+02/Frog2 (1.18 %, 3/253) in Se. khawi, and Trypanosoma Anura04/Frog1 (8.70 %, 22/253) in Se. khawi, Se. hivernus and Grossomyia indica. Bloodmeal analysis utilizing the COI gene revealed a diverse range of vertebrate hosts' blood, including bird, bat, frog and sun skink. Our findings confirm the presence of Trypanosoma DNA and identify the sources of bloodmeals from vertebrate hosts in various sand fly species, suggesting their potential as possible vectors for Trypanosoma in Thailand. Furthermore, our study is the first to provide molecular evidence using the COI gene to identify frogs as a host blood source for sand flies in Thailand. Further studies focusing on the isolation of live parasites in sand flies to confirm vector potential and examining the role of animal reservoirs will enhance our understanding of the host-parasite relationship and enable more efficient control for disease transmission.
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ADN Protozoario , Leishmania , Psychodidae , Trypanosoma , Animales , Tailandia/epidemiología , Trypanosoma/genética , Trypanosoma/aislamiento & purificación , Trypanosoma/clasificación , Femenino , Leishmania/genética , Leishmania/aislamiento & purificación , Leishmania/clasificación , ADN Protozoario/genética , Psychodidae/parasitología , Insectos Vectores/parasitología , Filogenia , Reacción en Cadena de la Polimerasa , ADN Espaciador Ribosómico/genética , Complejo IV de Transporte de Electrones/genética , Sangre/parasitologíaRESUMEN
Angiostrongylus vasorum is a metastrongylid parasite infecting wild canids and domestic dogs. Its patchy distribution, high pathogenicity and taxonomical classification makes the evolutionary history of A. vasorum intriguing and important to study. First larval stages of A. vasorum were recovered from feces of two grey foxes, Urocyon cinereoargenteus, from Costa Rica. Sequencing and phylogenetic and haplotypic analyses of the ITS2, 18S and cytochrome oxidase subunit 1 (cox1) fragments were performed. Then p- and Nei´s genetic distance, nucleotide substitution rates and species delimitation analyses were conducted with cox1 data of the specimens collected herein and other Angiostrongylus spp. Cophylogenetic congruence and coevolutionary events of Angiostrongylus spp. and their hosts were evaluated using patristic and phenetic distances and maximum parsimony reconciliations. Specimens from Costa Rica clustered in a separate branch from European and Brazilian A. vasorum sequences in the phylogenetic and haplotype network analyses using the ITS2 and cox1 data. In addition, cox1 p-distance of the sequences derived from Costa Rica were up to 8.6 % different to the ones from Europe and Brazil, a finding mirrored in Nei´s genetic distance PCoA. Species delimitation analysis supported a separate group with the sequences from Costa Rica, suggesting that these worms may represent cryptic variants of A. vasorum, a new undescribed taxon or Angiocaulus raillieti, a synonym species of A. vasorum described in Brazil. Moreover, nucleotide substitution rates in A. vasorum were up to six times higher than in the congener Angiostrongylus cantonensis. This finding and the long time elapsed since the last common ancestor between both species may explain the larger diversity in A. vasorum. Finally, cophylogenetic congruence was observed between Angiostrongylus spp. and their hosts, with cospeciation events occurring at deeper taxonomic branching of host order. Altogether, our data suggest that the diversity of the genus Angiostrongylus is larger than expected, since additional species may be circulating in wild canids from the Americas.