RESUMEN
Echinochasmids are a group of globally distributed digeneans, and the adults are found in the intestines of birds, mammals and reptiles. In the Neotropical region of Mexico, adult specimens were obtained from seven fish-eating bird species in six localities, whereas specimens of Stephanoprora aylacostoma were obtained experimentally in Argentina. Morphologically, the new specimens from the Neotropical region of Mexico were identified as Stephanoprora uruguayense and an undescribed species of Echinochasmus. Sequences for two nuclear (large subunit (28S) and internal transcribed spacer from DNA ribosomal (ITS1-5.8S-ITS2)) molecular markers were generated and analysed together with other sequences downloaded from GenBank. The phylogenies obtained with each molecular marker indicated that Echinochasmus is paraphyletic and agreed with previous phylogenetic studies. The first cluster included the type species (E. coaxatus, which has 24 head-collar spines) plus three congeneric species. The second cluster contained species of Echinochasmus plus Stephanoprora, including the species analysed herein, S. uruguayense, S. aylacostoma (with 22 head-collar spines) and Echinochasmus sp. (with 20 head-collar spines), which formed three independent subclades, allowing us to recognize a lineage that was described morphologically as a new species. Echinochasmus ostrowskiae n. sp. can be distinguished from its congeners by having a head collar with 20 spines in a single row, seven spines on each edge and three angle spines, and a pharynx with an irregular edge and by the body, egg and collar spine sizes. Additionally, new host and locality records for S. uruguayense are presented, expanding its geographical distribution range in the Americas.
Asunto(s)
Filogenia , Infecciones por Trematodos , Animales , México , Infecciones por Trematodos/parasitología , Infecciones por Trematodos/veterinaria , Trematodos/clasificación , Trematodos/anatomía & histología , Trematodos/genética , Trematodos/aislamiento & purificación , Aves/parasitología , ADN Espaciador Ribosómico/análisis , Enfermedades de las Aves/parasitología , Argentina , ADN de Helmintos/análisis , ARN Ribosómico 28S/análisis , ARN Ribosómico 28S/genéticaRESUMEN
The planktonic dinoflagellate Prorocentrum compressum is widespread in warm and temperate seas. A strain identified as P. cf. compressum BEA 0681B isolated from the island of Gran Canaria, NE Atlantic Ocean, showed a divergence in rDNA/ITS phylogenies with respect to P. compressum. The Canarian strain was oval, with an average length-to-width ratio of 1.35, smooth thecal surface with less than 150 thecal pores, including oblique pores, sometimes with a bifurcated opening. In contrast, P. compressum was rounder, with a length-to-width ratio < 1.2, with reticulate-foveate ornamentation and 200-300 pores per valve. We propose Prorocentrum canariense sp. nov. These species clustered as the most early-branching lineage in the clade Prorocentrum sensu stricto. Although this clade mainly contains planktonic species, the closer relatives were the benthic species P. tsawwassenense and P. elegans. Interestingly, P. compressum and P. canariense sp. nov. are widely distributed in temperate and warm seas without an apparent morphological adaptation to planktonic life. The formation of two concentric hyaline mucilaginous walls could contribute to this success. We discuss the use of Prorocentrum bidens to solve the nomenclature issue of P. compressum that was described citing a diatom as basionym.
Asunto(s)
ADN Protozoario , ADN Ribosómico , Dinoflagelados , Filogenia , Dinoflagelados/clasificación , Dinoflagelados/genética , ADN Ribosómico/genética , ADN Protozoario/genética , Océano Atlántico , Análisis de Secuencia de ADN , ADN Espaciador Ribosómico/genética , ADN Espaciador Ribosómico/análisis , Datos de Secuencia MolecularRESUMEN
Species of the genus Hysterothylacium are aquatic roundworms (nematodes) belonging to the family Raphidascarididae. Some species in this family are known to be associated with zoonotic diseases in humans after they consume their parasitic larvae in raw or undercooked fish. The aim of this research was to report the prevalence, morphology, and molecular characteristics of Hysterothylacium species in Pagellus erythrinus. A total of Two hundred fish were purchased from the fish market in Damanhour, Beheira Province, between December 2021 and November 2022 and subjected to examination. For molecular characterization, the internal transcribed spacer (ITS) region of nuclear ribosomal DNA and the mitochondrial cytochrome oxidase subunit 2 (COX-2) gene were used. Hysterothylacium species were morphologically described and identified from the intestine of Pagellus erythrinus in Beheira Province, Egypt. The PCR amplified 1087 bp and 629 bp of the target sequences of the ITS region and COX-2 gene, respectively. Sequence analysis revealed the Hysterothylacium thalassini species. The identified species provided novel biological data for the Hysterothylacium nematode in Pagellus erythrinus. The prevalence of Hysterothylacium species recovered from the intestine was 55%. The highest prevalence of 72% has been reported in summer compared to the lowest prevalence of 38% in the winter. Females had a higher prevalence of 61.8% than males, with 44.2%. The first detection, prevalence, and molecular characterization of H. thalassini in Pagellus erythrinus from Beheira Province, Egypt, was presented in this study.
Asunto(s)
Enfermedades de los Peces , Animales , Egipto/epidemiología , Enfermedades de los Peces/parasitología , Enfermedades de los Peces/epidemiología , Prevalencia , Mar Mediterráneo/epidemiología , Femenino , Masculino , Infecciones por Ascaridida/veterinaria , Infecciones por Ascaridida/parasitología , Infecciones por Ascaridida/epidemiología , Filogenia , Ascaridoidea/aislamiento & purificación , Ascaridoidea/genética , Ascaridoidea/clasificación , Complejo IV de Transporte de Electrones/análisis , Complejo IV de Transporte de Electrones/genética , ADN Espaciador Ribosómico/análisis , ADN Espaciador Ribosómico/genética , ADN de Helmintos/análisisRESUMEN
Fungi contribute to different important ecological processes, including decomposition of organic matter and nutrient cycling, but in the marine environment the main factors influencing their diversity and dynamics at the spatial and temporal levels are still largely unclear. In this study, we performed DNA metabarcoding on seawater sampled monthly over a year and a half in the Gulf of Trieste (northern Adriatic Sea), targeting the internal transcribed spacer (ITS) and the 18S rRNA gene regions. The fungal communities were diverse, very dynamic, and belonged predominantly to marine taxa. Samples could be clustered in two groups, mainly based on the high (> 30%) or low relative proportion of the ascomycetes Parengyodontium album, which emerged as a key taxon in this area. Dissolved and particulate organic C:N ratio played important roles in shaping the mycoplankton assemblages, suggesting that differently bioavailable organic matter pools may be utilized by different consortia. The proportion of fungal over total reads was 31% for ITS and 0.7% for 18S. ITS had the highest taxonomic resolution but low power to detect early divergent fungal lineages. Our results on composition, distribution, and environmental drivers extended our knowledge of the structure and function of the mycobiome of coastal waters.
Asunto(s)
Biodiversidad , Hongos , ARN Ribosómico 18S , Agua de Mar , Agua de Mar/microbiología , Hongos/genética , Hongos/clasificación , Hongos/aislamiento & purificación , ARN Ribosómico 18S/genética , ARN Ribosómico 18S/análisis , Micobioma , ADN de Hongos/genética , Código de Barras del ADN Taxonómico , Filogenia , ADN Espaciador Ribosómico/genética , ADN Espaciador Ribosómico/análisis , Ascomicetos/genética , Ascomicetos/clasificación , Ascomicetos/aislamiento & purificaciónRESUMEN
Vector control remains one of the best strategies to prevent the transmission of trypanosome infections in humans and livestock and, thus, a good way to achieve the elimination of human African trypanosomiasis and animal African trypanosomiasis. A key prerequisite for the success of any vector control strategy is the accurate identification and correct mapping of tsetse species. In this work, we updated the tsetse fly species identification and distribution in many geographical areas in Cameroon. Tsetse flies were captured from six localities in Cameroon, and their species were morphologically identified. Thereafter, DNA was extracted from legs of each tsetse fly and the length polymorphism of internal transcribed spacer-1 (ITS1) region of each fly was investigated using PCR. ITS1 DNA fragments of each tsetse species were sequenced. The sequences obtained were analysed and compared to those available in GenBank. This enabled to confirm/infirm results of the morphologic identification and then, to establish the phylogenetic relationships between tsetse species. Morphologic features allowed to clearly distinguish all the tsetse species captured in the South Region of Cameroon, that is, Glossina palpalis palpalis, G. pallicera, G. caliginea and G. nigrofusca. In the northern area, G. morsitans submorsitans could also be distinguished from G. palpalis palpalis, G. tachinoides and G. fuscipes, but these three later could not be distinguished with routine morphological characters. The ITS1 length polymorphism was high among most of the studied species and allowed to identify the following similar species with a single PCR, that is, G. palpalis palpalis with 241 or 242 bp and G. tachinoides with 221 or 222 bp, G. fuscipes with 236 or 237 bp. We also updated the old distribution of tsetse species in the areas assessed, highlighting the presence of G. palpalis palpalis instead of G. fuscipes in Mbakaou, or in sympatry with G. morsitans submorsitans in Dodeo (northern Cameroon). This study confirms the presence of G. palpalis palpalis in the Adamawa Region of Cameroon. It highlights the limits of using morphological criteria to differentiate some tsetse species. Molecular tools based on the polymorphism of ITS1 of tsetse flies can differentiate tsetse species through a simple PCR before downstream analyses or vector control planning.
Asunto(s)
Insectos Vectores , Polimorfismo Genético , Moscas Tse-Tse , Animales , Camerún , Moscas Tse-Tse/genética , Insectos Vectores/genética , Insectos Vectores/clasificación , Distribución Animal , Filogenia , ADN Intergénico/genética , Femenino , Control de Insectos , Masculino , ADN Espaciador Ribosómico/análisis , ADN Espaciador Ribosómico/genética , Análisis de Secuencia de ADNRESUMEN
The growing concern about migratory birds potentially spreading ticks due to global warming has become a significant issue. The city of Nantong in this study is situated along the East Asia-Australasian Flyway (EAAF), with numerous wetlands serving as roosting sites for migratory birds. We conducted an investigation of hard ticks and determined the phylogenetic characteristics of tick species in this city. We utilized three different genes for our study: the mitochondrial cytochrome oxidase subunit 1 (COX1) gene, the second internal transcribed spacer (ITS2), and the mitochondrial small subunit rRNA (12 S rRNA) gene. The predominant tick species were Haemaphysalis flava (H. flava) and Haemaphysalis longicornis (H. longicornis). Additionally, specimens of Haemaphysalis campanulata (H. campanulata) and Rhipicephalus sanguineus (R. sanguineus) were collected. The H. flava specimens in this study showed a close genetic relationship with those from inland provinces of China, as well as South Korea and Japan. Furthermore, samples of H. longicornis exhibited a close genetic relationship with those from South Korea, Japan, Australia, and the USA, as well as specific provinces in China. Furthermore, R. sanguineus specimens captured in Nantong showed genetic similarities with specimens from Egypt, Nigeria, and Argentina.
Asunto(s)
Migración Animal , Aves , Complejo IV de Transporte de Electrones , Ixodidae , Filogenia , Animales , China , Ixodidae/genética , Ixodidae/clasificación , Ixodidae/fisiología , Complejo IV de Transporte de Electrones/genética , Complejo IV de Transporte de Electrones/análisis , ARN Ribosómico/genética , ARN Ribosómico/análisis , Ninfa/crecimiento & desarrollo , Ninfa/clasificación , Ninfa/genética , Ninfa/fisiología , Proteínas de Artrópodos/genética , Proteínas de Artrópodos/análisis , ADN Espaciador Ribosómico/análisisRESUMEN
Three new species of Gyrodactylus are described from three species of bitterling in Donghu Lake, China: Gyrodactylus ocellorhodei n. sp. from Rhodeus ocellatus; G. sinenorhodei n. sp. from Rhodeus sinensis; and G. acheilorhodei n. sp. from Acheilognathus macropterus. All the three new species showed similar opisthaptor morphology, especially the marginal hooks: all had a slender and perpendicular sickle shaft, and flat sickle base with distinct heel and inner arch which was different from the G. rhodei-group species parasitic on bitterling. Multivariate analyses based on hamulus and marginal hooks suggested that these three new species cannot be completely distinguished, despite some morphology divergence observed in certain less reliable morphometric features, such as hamulus root length, ventral bar total length and process shape. These three new species shared an identical 18S ribosomal RNA gene sequence, while the variation in the Internal Transcribed Spacers (ITS1-ITS2) sequence among them (8.4-11.2%, K2P) far exceeded the 1% ITS sequence difference that had been suggested as a threshold for species delimitation of Gyrodactylus. Phylogenetic analysis based on ITS1-ITS2 showed that all these sequenced Gyrodactylus spp. parasitic on the subfamily Acheilognathinae host formed a monophyletic group. However, a clear differentiation (18.9-20.9%, K2P of ITS1-ITS2) could be found between the subgroup from China (G. ocellorhodei n. sp., G. sinenorhodei n. sp. and G. acheilorhodei n. sp.) and that from Europe (G. rhodei).
Asunto(s)
Enfermedades de los Peces , Filogenia , Trematodos , Infecciones por Trematodos , Animales , Enfermedades de los Peces/parasitología , China , Infecciones por Trematodos/parasitología , Infecciones por Trematodos/veterinaria , Trematodos/clasificación , Trematodos/anatomía & histología , Trematodos/genética , Trematodos/aislamiento & purificación , ARN Ribosómico 18S/análisis , Cyprinidae/parasitología , ADN Espaciador Ribosómico/análisis , ADN de Helmintos/análisis , Lagos/parasitología , Platelmintos/clasificación , Platelmintos/anatomía & histología , Platelmintos/aislamiento & purificación , Platelmintos/genéticaRESUMEN
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.
Asunto(s)
Enfermedades de los Peces , Trematodos , Infecciones por Trematodos , Zooplancton , Animales , Australia , Cnidarios/clasificación , ADN de Helmintos/análisis , ADN Mitocondrial/análisis , ADN Ribosómico/análisis , ADN Espaciador Ribosómico/análisis , Enfermedades de los Peces/parasitología , Enfermedades de los Peces/epidemiología , Peces/parasitología , Japón , Metacercarias/aislamiento & purificación , Filogenia , Trematodos/clasificación , Trematodos/genética , Trematodos/aislamiento & purificación , Trematodos/anatomía & histología , Infecciones por Trematodos/veterinaria , Infecciones por Trematodos/parasitología , Infecciones por Trematodos/epidemiologíaRESUMEN
Mud volcanoes (MVs) are visible signs of oil and gas reserves present deep beneath land and sea. The Marac MV in Trinidad is the only MV associated with natural hydrocarbon seeps. Petrogenic polyaromatic hydrocarbons (PAHs) in its sediments must undergo biogeochemical cycles of detoxification as they can enter the water table and aquifers threatening ecosystems and biota. Recurrent hydrocarbon seep activity of MVs consolidates the growth of hydrocarbonoclastic fungal communities. Fungi possess advantageous metabolic and ecophysiological features for remediation but are underexplored compared to bacteria. Additionally, indigenous fungi are more efficient at PAH detoxification than commercial/foreign counterparts and remediation strategies remain site-specific. Few studies have focused on hydrocarbonoclastic fungal incidence and potential in MVs, an aspect that has not been explored in Trinidad. This study determined the unique biodiversity of culturable fungi from the Marac MV capable of metabolizing PAHs in vitro and investigated their extracellular peroxidase activity to utilize different substrates ergo their extracellular oxidoreductase activity (> 50% of the strains decolourized of methylene blue dye). Dothideomycetes and Eurotiomycetes (89% combined incidence) were predominantly isolated. ITS rDNA sequence cluster analysis confirmed strain identities. 18 indigenous hydrocarbonoclastic strains not previously reported in the literature and some of which were biosurfactant-producing, were identified. Intra-strain variability was apparent for PAH utilization, oil-tolerance and hydroxylase substrate specificity. Comparatively high levels of extracellular protein were detected for strains that demonstrated low substrate specificity. Halotolerant strains were also recovered which indicated marine-mixed substrata of the MV as a result of deep sea conduits. This work highlighted novel MV fungal strains as potential bioremediators and biocatalysts with a broad industrial applications.
Asunto(s)
Biotransformación , Hongos/aislamiento & purificación , Hongos/metabolismo , Hidrocarburos Policíclicos Aromáticos/metabolismo , Biodiversidad , ADN de Hongos/análisis , ADN Ribosómico/análisis , ADN Espaciador Ribosómico/análisis , Enzimas , Hongos/enzimología , Sedimentos Geológicos/microbiología , Peroxidasa , Petróleo , Salinidad , Análisis de Secuencia de ADN , Trinidad y TobagoRESUMEN
The superfamily Opisthorchioidea encompasses the families Cryptogonimidae, Opisthorchiidae and Heterophyidae. These parasites depend on the aquatic environment and include marine and freshwater species. Some species, such as Clonorchis sinensis and Opisthorchis viverrini, have a high impact on public health with millions of infected people worldwide and have thus been the object of many studies and tool developments. However, for many species, tools for identification and detection are scarce. Although morphological descriptions have been used and are still important, they are often not efficient on the immature stages of these parasites. Thus, during the past few decades, molecular approaches for parasite identification have become commonplace. These approaches are efficient, quick and reliable. Nonetheless, for some parasites of the superfamily Opisthorchioidea, reference genomic data are limited. This study reviews available genetic data and molecular tools for the identification and/or the detection of this superfamily. Molecular data on this superfamily are mostly based on mitochondrial and ribosomal gene sequence analyses, especially on the cytochrome c oxidase subunit 1 gene and internal transcribed spacer regions respectively.
Asunto(s)
ADN de Helmintos/genética , Parasitología/métodos , Trematodos/clasificación , Animales , Cartilla de ADN , ADN de Helmintos/análisis , ADN Mitocondrial/genética , ADN Ribosómico/genética , ADN Espaciador Ribosómico/análisis , Complejo IV de Transporte de Electrones/genética , Genes de Helminto , Heterophyidae/clasificación , Heterophyidae/genética , Heterophyidae/aislamiento & purificación , Técnicas de Diagnóstico Molecular , Técnicas de Amplificación de Ácido Nucleico , Opisthorchidae/clasificación , Opisthorchidae/genética , Opisthorchidae/aislamiento & purificación , Filogenia , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Técnica del ADN Polimorfo Amplificado Aleatorio , Trematodos/genética , Trematodos/aislamiento & purificaciónRESUMEN
Paradiplozoon opsariichthydis (Jiang, Wu et Wang, 1984) Jiang, Wu et Wang, 1989 (Platyhelminthes, Monogenea, Diplozoidae) is blood-feeding parasite from the gills of Asian cyprinid fish Opsariichthys bidens Günther, 1873. In this study, we present a morphological redescription of P. opsariichthydis neotype main morphological features e.g. size of body and clamps due to the fact that the type material is missing. We decided to supplement morphological descriptions by the relevant molecular data (internal transcribed spacer - ITS2) related to P. opsariichthydis adult worm isolates and other representatives of genus Paradiplozoon to cross verify our findings. In addition to that, this study also brings an attention to the host identification. Thus, parasite data were complemented by the determinant cytochrome oxidase b (cytb) sequences of its hosts. All novel sequences are deposited in GenBank. This combination of the morphological and molecular data related to both the parasite and its host seems to be the optimal approach to the general process of (re)description of highly host-specific parasitic organisms, which can then lead to a meaningful phylogenetic analysis.
Asunto(s)
Cyprinidae , Interacciones Huésped-Parásitos , Filogenia , Trematodos/anatomía & histología , Animales , Citocromos b/análisis , ADN de Helmintos/análisis , ADN Espaciador Ribosómico/análisis , Femenino , Enfermedades de los Peces/parasitología , Proteínas de Peces/análisis , Masculino , Trematodos/clasificación , Trematodos/genética , Infecciones por Trematodos/parasitologíaRESUMEN
Wild animals often act as reservoirs of tick-borne Babesia and Theileria spp., which cause piroplasmosis. Therefore, epidemiological investigations about the distribution of these parasites in wild animals are important for evaluating the transmission risk to humans and livestock. In this study, we surveyed Babesia and Theileria spp. infecting wild boar (Sus scrofa) in Kagoshima and Yamaguchi prefectures and Tsushima island, which are all in western Japan, and performed molecular genetic analyses on the samples. DNA was extracted from either blood or liver samples of wild boar captured in Kagoshima prefecture in 2015, 2016, and 2018 and from blood samples from wild boar captured in Yamaguchi prefecture in 2013-2015 and Tsushima island in 2018. PCR screening for the partial 18S ribosomal RNA gene (18S rRNA) of both Babesia and Theileria spp. in wild boar revealed that 63.9 % (140 of 219 samples) were positive. Sequencing of all positive samples revealed that they were all the same Babesia species. Subsequent phylogenetic analyses showed that the parasite is closely related to Babesia sp. previously detected in the hard tick, Amblyomma testudinarium in Kagoshima, and further analyses suggested that this species is genetically related to Babesia gibsoni. On the other hand, no Theileria were detected in any of the samples. In summary, we observed a high prevalence of B. gibsoni-like Babesia sp. in wild boar in western regions of Japan. The host range, distribution, pathogenicity, and life cycle of this protozoan should be further evaluated.
Asunto(s)
Babesia/aislamiento & purificación , Babesiosis/epidemiología , Enfermedades de los Porcinos/epidemiología , Animales , Babesia/genética , Babesiosis/parasitología , Citocromos b/análisis , ADN Protozoario/análisis , ADN Espaciador Ribosómico/análisis , Japón/epidemiología , Filogenia , Prevalencia , Proteínas Protozoarias/análisis , ARN Protozoario/análisis , ARN Ribosómico 18S/análisis , Sus scrofa , Porcinos , Enfermedades de los Porcinos/parasitologíaRESUMEN
BACKGROUND: The malaria mosquito Anopheles punctipennis, a widely distributed species in North America, is capable of transmitting human malaria and is actively involved in the transmission of the ungulate malaria parasite Plasmodium odocoilei. However, molecular diagnostic tools based on Internal Transcribed Spacer 2 (ITS2) of ribosomal DNA are lacking for this species. Anopheles punctipennis is a former member of the Anopheles maculipennis complex but its systematic position remains unclear. METHODS: In this study, ITS2 sequences were obtained from 276 An. punctipennis specimens collected in the eastern and midwestern United States and a simple and robust Restriction Fragment Length Polymorphism approach for species identification was developed. The maximum-likelihood phylogenetic tree was constructed based on ITS2 sequences available through this study and from GenBank for 20 species of Anopheles. RESULTS: The analysis demonstrated a consistent ITS2 sequence length and showed no indications of intragenomic variation among the samples based on ITS2, suggesting that An. punctipennis represents a single species in the studied geographic locations. In this study, An. punctipennis was found in urban, rural, and forest settings, suggesting its potential broad role in pathogen transmission. Phylogeny based on ITS2 sequence comparison demonstrated the close relationship of this species with other members of the Maculipennis group. CONCLUSIONS: This study developed molecular tools based on ITS2 sequences for the malaria vector An. punctipennis and clarified the phylogenetic position of the species within the Maculipennis group.
Asunto(s)
Distribución Animal , Anopheles/clasificación , ADN Espaciador Ribosómico/análisis , Mosquitos Vectores/clasificación , Polimorfismo de Longitud del Fragmento de Restricción , Animales , Anopheles/genética , Anopheles/fisiología , Florida , Iowa , Malaria/transmisión , Minnesota , Mosquitos Vectores/genética , Mosquitos Vectores/fisiología , VirginiaRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: The roots and stems of several Salacia species have been used as traditional medicines, especially in Ayurvedic medical system for the treatment of diabetes, rheumatism, gonorrhea, amenorrhea, skin diseases, etc. Due to reported evidence supporting Salacia's beneficial effects in early-stage diabetes and other lifestyle-related diseases, Salacia-based dietary supplements and health foods have been gaining popularity in Japan and other countries in recent years. However, due to the morphological similarities between Salacia plants, particularly in the medicinally used parts (roots and stems), the authentication of the botanical identities of Salacia-derived products is challenging. AIM OF THIS STUDY: This study aims to develop a genetic approach to authenticate the medicinally used Salacia species and to determine the botanical sources of the commercially available Salacia-derived products. MATERIALS AND METHODS: The sequences of nuclear DNA internal transcribed spacer (ITS) and chloroplast trnK-rps16 region were determined and compared between 10 plant specimens from three medicinally used Salacia species as well as 48 samples of commercial crude drugs. Moreover, a PCR-restriction fragment length polymorphism (RFLP) assay was developed for rapid identification based on the ITS sequences. RESULTS: The plant specimens from the three medicinally used Salacia species showed three main types of sequences in both ITS (types I, II, III) and trnK-rps16 (i, ii, iii) regions. Combined the sequences of ITS and trnK-rps16 regions, S. reticulata and S. oblonga had type I-i and type III-iii or similar sequences, respectively. S. chinensis had type II-ii or II(536M)-i sequences. Forty-eight samples of commercial crude drugs were identified based on ITS and trnK-rps16 DNA barcode. A convenient PCR-RFLP assay using Cac8I restriction enzyme was established and applied to identify the botanical sources of health food products purchased from online retailers. All the twelve samples were identified as S. chinensis. CONCLUSION: The nrDNA ITS sequences provided useful information to authenticate Salacia species and to elucidate the phylogenetic relationship within the Salacia genus. Genetic identification results revealed that S. chinensis and S. reticulata are the major sources of commercially available Salacia-products. Based on the ITS sequences, a convenient PCR-RFLP assay was established for the identification of the medicinally used Salacia species as well as their derived health food products.
Asunto(s)
Código de Barras del ADN Taxonómico/métodos , ADN de Plantas/genética , ADN Espaciador Ribosómico/genética , ADN Espaciador Ribosómico/aislamiento & purificación , Reacción en Cadena de la Polimerasa/métodos , Salacia/clasificación , Salacia/genética , ADN de Cloroplastos/análisis , ADN de Cloroplastos/genética , ADN Espaciador Ribosómico/análisis , Suplementos Dietéticos/análisis , Análisis de los Alimentos , Filogenia , Polimorfismo de Longitud del Fragmento de RestricciónRESUMEN
Given that accurately identifying pathogen vectors is vital for designing efficient mosquito control programs based on the proper surveillance of the epidemiologically important species, it has been suggested the complementary use of independently evolving genes and morphometric traits as a reliable approach for the characterization and delimitation of related species. Hence, we examined the spatial distribution of COI mtDNA and ITS2 rDNA variation from the historical perspective of Ochlerotatus caspius (Pallas, 1771) and O. dorsalis (Meigen, 1830), while simultaneously testing the utility of the two markers in integrative species delimitation when combined with phenotypic character analyses of larvae and adults. Despite the striking difference in haplotype diversity (high in COI mtDNA, low in ITS2 rDNA), no evident phylogeographic structure was apparent in the Palearctic O. caspius. The Holarctic O. dorsalis species was subdivided into two highly distinctive COI mtDNA phylogroups which corresponded to the Nearctic and Palearctic regions. Strong support for the independence of the two allopatric evolutionary lineages suggested that geographical barrier and climatic changes during Pleistocene caused vicariance of the ancestral range. COI mtDNA reliably distinguished O. caspius and O. dorsalis, while ITS2 rDNA yet again lacked the proper resolution for solving this problem. An integrative approach based on the larval and adult morphological traits have varying taxonomic applications due to their differential diagnostic values. Thus, by the implementation of an integrative taxonomic approach, we successfully detected species borders between the two epidemiologically relevant species and uncovered the presence of cryptic diversity within O. dorsalis.
Asunto(s)
Variación Genética , Ochlerotatus/clasificación , Ochlerotatus/genética , Animales , ADN Espaciador Ribosómico/análisis , Complejo IV de Transporte de Electrones/análisis , Femenino , Marcadores Genéticos/genética , Haplotipos , Proteínas de Insectos/análisis , Larva/clasificación , Larva/enzimología , Larva/genética , Masculino , Ochlerotatus/enzimología , Filogeografía , Especificidad de la EspecieRESUMEN
Cutaneous leishmaniasis (CL) is a major health problem in Iran, with a heavy burden on human health and society. There is little knowledge about the molecular epidemiology of the disease, as well as phylogenetic relationship of causative agents in south-eastern Iran. The aim of the present study was to investigate the molecular aspects of CL, especially atypical CL in the Bam district, Kerman province, south-eastern Iran, as an endemic region of CL in Iran. The smears were collected from lesion samples of 353 patients clinically suspected to CL, who attended local health centres in the Bam district during 2016-2017. Direct smears were examined for Leishmania parasites using the Giemsa staining technique. Amplification of kinetoplast DNA (kDNA) and the ribosomal internal transcribed spacer 1(ITS-1) gene were carried out using polymerase chain reaction (PCR). Then, the ITS1-PCR products were sequenced for phylogenetic analysis. Overall, 278 cases were confirmed as CL by microscopic examination of Giemsa-stained slides. Clinical presentation of the lesions was basically of two types: (a) typical lesions and (b) atypical including lupoid ulcers, sporotrichoid, nodular and exudative lesions. The PCR assay on all specimens of skin lesions proved L. tropica as the main pathogenic agent. Phylogenic analysis revealed high similarity among isolates from the Bam district in the south-east with isolates from Birjand in eastern Iran, as well as with isolates from Herat province in western Afghanistan. The study provided valuable information concerning the genetic diversity of the parasite as one of the factors influencing the clinical manifestations in CL in south-eastern Iran, which could be the basis for planning future control strategies.
Asunto(s)
Leishmania tropica/aislamiento & purificación , Leishmaniasis Cutánea/parasitología , Adolescente , Adulto , Anciano , Niño , Preescolar , ADN de Cinetoplasto/análisis , ADN Espaciador Ribosómico/análisis , Femenino , Humanos , Lactante , Irán , Leishmania tropica/clasificación , Leishmania tropica/genética , Masculino , Persona de Mediana Edad , Filogenia , Adulto JovenRESUMEN
Extralymphatic filariasis is an uncommon phenomenon that can be caused by several lymphatic filarial species, including zoonotic filaria of animal origins. In this study, we report a case of a 64-year-old Thai woman who presented with a lump in her left breast that was diagnosed with invasive ductal carcinoma. At the same time, a small nodule was found in her right breast, via imaging study, without any abnormal symptoms. A core needle biopsy of the right breast nodule revealed a filarial-like nematode compatible with the adult stage of Brugia sp. A molecular identification of the nematode partial mt 12rRNA gene and ITS1 suggested the causative species as closely related to Brugia pahangi, a zoonotic lymphatic filaria of animals such as cats and dogs. The sequence of the partial mt 12rRNA and ITS1 gene in this patient was 94% and 99% identical to the previously reported sequence of mt 12rRNA and ITS1 genes of B. pahangi. The sequence of ITS1 gene is 99% similar to B. pahangi microfilaria from infected dogs in Bangkok, which was highly suspected of having a zoonotic origin. As far as we know, this is the first case report of B. pahangi filariasis presented with a breast mass concomitantly found in a patient with invasive ductal carcinoma. This raised serious concern regarding the zoonotic transmission of filariasis from natural animal reservoirs.
Asunto(s)
Enfermedades de la Mama/diagnóstico , Neoplasias de la Mama/patología , Brugia pahangi/aislamiento & purificación , Carcinoma Ductal de Mama/patología , Filariasis/diagnóstico , Animales , Enfermedades de la Mama/parasitología , Brugia pahangi/clasificación , ADN Espaciador Ribosómico/análisis , Femenino , Filariasis/parasitología , Humanos , Persona de Mediana Edad , ARN de Helminto/análisis , ARN Ribosómico/análisis , TailandiaRESUMEN
Females, nymphs, and larvae of Ixodes silvanus n. sp. collected from birds and from the vegetation in northwestern Argentina (Yungas Phytogeographic Province) are described herein. The new species belongs to the subgenus Trichotoixodes (Acari: Ixodidae). The female is diagnosed by a combination of the following characters: scutum with setae moderately long and more numerous in central field, fewer and moderately long setae on lateral fields, and inconspicuous setae in anterior field; basis capituli subtriangular dorsally; porose areas large and irregular in shape, lacking distinct margins; auriculae with straight edges diverging posterolaterally and ending with small blunt processes; hypostome narrow and pointed with dental formula 4/4 in the anterior third, then 3/3 and 2/2 near the base; coxae I with two spurs, sub-equal in size, internal slightly slimmer than external. The nymph is diagnosed by notum with numerous and long setae, ventral surface covered by numerous whitish setae, scutum with short scapulae and few and shallow punctations, setae on scutum few, short and irregularly distributed, basis capituli sub-triangular dorsally with posterior margin straight, cornua large and directed postero-laterally, auriculae large and projected laterally, lateral margin of basis capituli above auriculae with a lateral and triangular projection, hypostome pointed with dental formula 3/3 in the anterior third and then 2/2, and coxa I with two short, sub-equal, triangular spurs. The diagnostic characters of the larva are: basis capituli dorsally sub-triangular with lateral angles acute and posterior margin straight, auriculae as large triangular lateral projections, hypostome with apex bluntly pointed and dental formula 3/3 in the anterior third and then 2/2, coxa I with two short, sub-equal, triangular spurs, and pattern of dorsal and ventral body setae. This new species is phylogenetically related to Ixodes brunneus, Ixodes turdus and Ixodes frontalis, and the principal hosts for all its parasitic stages are birds.
Asunto(s)
Ixodes/anatomía & histología , Ixodes/clasificación , Animales , Argentina , ADN Espaciador Ribosómico/análisis , Complejo IV de Transporte de Electrones/análisis , Femenino , Ixodes/crecimiento & desarrollo , Ixodes/ultraestructura , Larva/anatomía & histología , Larva/clasificación , Larva/crecimiento & desarrollo , Larva/ultraestructura , Microscopía , Microscopía Electrónica de Rastreo , Ninfa/anatomía & histología , Ninfa/clasificación , Ninfa/crecimiento & desarrollo , Ninfa/ultraestructura , Filogenia , ARN Ribosómico 16S/análisisRESUMEN
Amblyomma cajennense Fabricius, 1787 (Acari: Ixodidae) is a widely distributed tick taxon. Recent studies have reassessed this taxon as a complex of six species. Amblyomma mixtum Koch, 1844 has been suggested by some authors as the only species of this complex that is present in Cuba. Other authors have pointed a niche overlapping for A. mixtum and A. cajennense s.s. in the country. Detailed taxonomic studies on the Cuban species belonging to this complex are needed in order to evaluate their current distribution according to the recent classification. This study aimed to characterize Cuban populations from the A. cajennense complex by using tick samples obtained from 3 occidental provinces and 1 central province of the country. Morphological identification and measurements of the main relevant taxonomic structures were conducted by using Scanning Electron Microscopy. Phylogenetic analyzes were carried out with 16S ribosomal RNA, internal transcribed spacer 2 and the subunit I of mitochondrial cytochrome c oxidase gene sequences. The results of these studies demonstrated that all samples belonged to the species A. mixtum (Koch, 1844). This study constitutes the first molecular characterization of this Amblyomma species in Cuba. Further studies will be necessary in order to corroborate if A. cajennense s.s. is also present in the island.
Asunto(s)
Amblyomma/anatomía & histología , Amblyomma/genética , Distribución Animal , Amblyomma/crecimiento & desarrollo , Animales , Cuba , ADN Espaciador Ribosómico/análisis , Perros/parasitología , Complejo IV de Transporte de Electrones/análisis , Femenino , Caballos/parasitología , Larva/anatomía & histología , Larva/genética , Larva/crecimiento & desarrollo , Masculino , Ninfa/anatomía & histología , Ninfa/genética , Ninfa/crecimiento & desarrollo , Filogenia , ARN Ribosómico 16S/análisis , Oveja Doméstica/parasitologíaRESUMEN
Echidnophaga gallinacea is the sticktight flea of chickens. It causes dermatitis and ulcers in the skin and carries some disease-causing agents such as Rickettsia and Bartonella. This study was conducted to detect the infection rate and elucidate the molecular characterization of E. gallinacea in chickens from El-Dabaa City, Matrouh Governorate, Egypt. The fleas were collected from infected chickens and identified morphologically. The internal transcribed spacer-1 (ITS-1) gene PCR method was used for molecular characterization. Based on the morphology, the collected fleas were confirmed as E. gallinacea. The overall infection rate was 5%, with 4.5% in female and 10% in male chickens. ITS-1 PCR revealed a specific band of 488 bp. The ITS-1 gene sequence from Egypt occurred in the same phylogenetic clade as that from Cameroon, with a percentage identity of 98.47%.