RESUMO
Homologous recombination (HR) is an essential DNA double-strand break (DSB) repair mechanism, which is frequently inactivated in cancer. During HR, RAD51 forms nucleoprotein filaments on RPA-coated, resected DNA and catalyzes strand invasion into homologous duplex DNA. How RAD51 displaces RPA and assembles into long HR-proficient filaments remains uncertain. Here, we employed single-molecule imaging to investigate the mechanism of nematode RAD-51 filament growth in the presence of BRC-2 (BRCA2) and RAD-51 paralogs, RFS-1/RIP-1. BRC-2 nucleates RAD-51 on RPA-coated DNA, whereas RFS-1/RIP-1 acts as a "chaperone" to promote 3' to 5' filament growth via highly dynamic engagement with 5' filament ends. Inhibiting ATPase or mutation in the RFS-1 Walker box leads to RFS-1/RIP-1 retention on RAD-51 filaments and hinders growth. The rfs-1 Walker box mutants display sensitivity to DNA damage and accumulate RAD-51 complexes non-functional for HR in vivo. Our work reveals the mechanism of RAD-51 nucleation and filament growth in the presence of recombination mediators.
Assuntos
Proteínas de Caenorhabditis elegans/genética , Caenorhabditis elegans/genética , Proteínas de Transporte/genética , DNA de Helmintos/genética , Proteínas de Ligação a DNA/genética , Rad51 Recombinase/genética , Reparo de DNA por Recombinação , Animais , Caenorhabditis elegans/metabolismo , Proteínas de Caenorhabditis elegans/metabolismo , Proteínas de Transporte/metabolismo , Quebras de DNA de Cadeia Dupla , DNA de Helmintos/metabolismo , Proteínas de Ligação a DNA/metabolismo , Regulação da Expressão Gênica , Chaperonas Moleculares/genética , Chaperonas Moleculares/metabolismo , Mutação , Ligação Proteica , Rad51 Recombinase/metabolismo , Proteína de Replicação A/genética , Proteína de Replicação A/metabolismo , Transdução de Sinais , Imagem Individual de MoléculaRESUMO
A new species of Moniliformis, M. tupaia n. sp. is described using integrated morphological methods (light and scanning electron microscopy) and molecular techniques (sequencing and analysing the nuclear 18S, ITS, 28S regions and mitochondrial cox1 and cox2 genes), based on specimens collected from the intestine of the northern tree shrew Tupaia belangeri chinensis Anderson (Scandentia: Tupaiidae) in China. Phylogenetic analyses show that M. tupaia n. sp. is a sister to M. moniliformis in the genus Moniliformis, and also challenge the systematic status of Nephridiacanthus major. Moniliformis tupaia n. sp. represents the third Moniliformis species reported from China.
Assuntos
Acantocéfalos , Filogenia , Tupaia , Animais , Tupaia/parasitologia , Tupaia/genética , China , Acantocéfalos/genética , Acantocéfalos/classificação , Acantocéfalos/anatomia & histologia , Acantocéfalos/ultraestrutura , Helmintíase Animal/parasitologia , Microscopia Eletrônica de Varredura/veterinária , DNA de Helmintos/genética , RNA Ribossômico 18S/genética , Feminino , Masculino , RNA Ribossômico 28S/genética , Intestinos/parasitologiaRESUMO
Rat lungworm disease or neuroangiostrongyliasis is a cerebral parasitic infection that affects humans and animals alike. Its clinical signs and symptoms can range from mild self-resolving to serious life-threatening conditions. Studies suggest therapeutic interventions during the early stages of infection to be more effective than in later stages. However, early diagnosis of infection is usually problematic without the knowledge of exposure and/or detection of the parasite's DNA or antibody against the parasite in the cerebrospinal fluid. This requires a lumbar puncture, which is an invasive procedure that generally requires hospitalization. This study evaluates an affordable and less invasive alternative to detect parasitic DNA by PCR from the peripheral blood of potentially infected animals. Blood samples from 58 animals (55 dogs and 3 cats) with clinical suspicion of infection were submitted to our lab between February 2019 and August 2022 by local, licensed veterinarians. DNA was extracted from whole blood, plasma, serum, and/or packed cells using the Qiagen DNeasy Blood & Tissue Kit as per the manufacturer's protocol. All 58 animals were tested by real-time PCR using the AcanITS1 assay and 32 of these animals (31dogs; 1 cat) were also tested using the AcanR3990 assay. The PCR results for both assays were classified into strongly positive > positive > weakly positive > negative, and equivocal for ambiguous results, based on the strength of the signal. The percent infection detected using the AcanITS1 and AcanR3990 assays was 12.72% (7/55) and 20.68% (6/29), respectively. The overall percent infection detected was 34.37% (11/32), with only two animals testing positive by both assays. The three cats involved in this study tested negative by both assays. These results are promising and warrant further investigations to increase sensitivity including variables that might affect detection in the blood, such as parasite load, and laboratory methodologies.
Assuntos
Angiostrongylus cantonensis , Doenças do Gato , Reação em Cadeia da Polimerase em Tempo Real , Infecções por Strongylida , Animais , Angiostrongylus cantonensis/isolamento & purificação , Angiostrongylus cantonensis/genética , Infecções por Strongylida/veterinária , Infecções por Strongylida/parasitologia , Infecções por Strongylida/diagnóstico , Infecções por Strongylida/sangue , Reação em Cadeia da Polimerase em Tempo Real/métodos , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Gatos , Doenças do Gato/parasitologia , Doenças do Gato/diagnóstico , Doenças do Gato/sangue , Cães , Doenças do Cão/parasitologia , Doenças do Cão/diagnóstico , Doenças do Cão/sangue , Sensibilidade e Especificidade , DNA de Helmintos/genética , DNA de Helmintos/sangueRESUMO
Echinococcus granulosus sensu lato (s.l.) is a species complex with the potential to cause cystic echinococcosis (CE). Contact with the feces of domestic dogs (Canis familiaris) fed with raw viscera of intermediate livestock hosts is a risk factor for this infection in the southern region of Brazil. Although the region has been considered endemic to CE for many years, molecular data regarding the species of the complex causing CE in humans are scarce. This study aimed to perform a molecular analysis of the biological fluid from a human liver cyst to investigate the species responsible for CE. Genetic material obtained from the hydatid fluid of a hepatic cyst from a human with CE was subjected to PCR to amplify mitochondrial and nuclear DNA sequences. The phylogenetic analysis confirmed the human infection by Echinococcus canadensis G7 in the state of Paraná, Brazil. This is the first molecular record of E. canadensis G7 infecting a human in Brazil, and it is important to reiterate the risk of human CE caused by this species in South America, as reported by a previous study in Patagonia, Argentina. From the epidemiological point of view, this finding is of great relevance for the southern region of Brazil, since this parasite has previously only been detected in pigs in the state of Rio Grande do Sul, neighboring Paraná. The finding points to the importance of this identification in the molecular epidemiology of E. granulosus s.l., especially in South America.
Assuntos
DNA de Helmintos , Echinococcus , Filogenia , Animais , Brasil/epidemiologia , Echinococcus/genética , Echinococcus/classificação , Echinococcus/isolamento & purificação , Humanos , DNA de Helmintos/genética , Equinococose/veterinária , Equinococose/parasitologia , Equinococose/epidemiologia , Análise de Sequência de DNA , Reação em Cadeia da Polimerase , DNA Mitocondrial/genética , MasculinoRESUMO
Mastophorus muris (Gmelin, 1790) is a globally distributed parasitic nematode of broad range mammals. The taxonomy within the genus Mastophorus and the cryptic diversity among the genus are controversial among taxonomists. This study provides a detailed morphological description of M. muris from Mus musculus combined with a molecular phylogenetic approach. Moreover, descriptions and molecular data of M. muris from non-Mus rodents and wildcats complement our findings and together provide new insights into their taxonomy. The analysis of M. muris was based on light microscopy and scanning electron microscopy. The morphological description focused on the dentition pattern of the two trilobed pseudolabia. Additionally, we described the position of the vulva, arrangement of caudal pairs of papillae, spicules and measured specimens from both sexes and the eggs. For the molecular phylogenetic approach, we amplified the small subunit ribosomal RNA gene and the internal transcribed spacer, and the cytochrome c oxidase subunit 1. Mastophorus morphotypes based on dentition patterns and phylogenetic clustering indicate a subdivision of the genus in agreement with their host. We recognize two groups without a change to formal taxonomy: One group including those specimens infecting Mus musculus, and the second group including organisms infecting non-Mus rodents. Our genetic and morphological data shed light into the cryptic diversity within the genus Mastopohorus. We identified two host-associated groups of M. muris. The described morphotypes and genotypes of M. muris allow a consistent distinction between host-associated parasites.
Assuntos
Microscopia Eletrônica de Varredura , Filogenia , Animais , Feminino , Masculino , Camundongos , Spiruroidea/classificação , Spiruroidea/genética , Spiruroidea/anatomia & histologia , Spiruroidea/isolamento & purificação , Spiruroidea/ultraestrutura , Complexo IV da Cadeia de Transporte de Elétrons/genética , Variação Genética , Análise de Sequência de DNA , Microscopia , DNA de Helmintos/genética , DNA Ribossômico/genética , DNA Espaçador Ribossômico/genética , Análise por Conglomerados , Dados de Sequência MolecularRESUMO
Among the species described within the Onchocercidae family, Dirofilaria immitis is regarded as the most common worldwide, causing severe and often fatal conditions in dogs, cats, and occasionally humans. Dirofilaria spp. are vectored by mosquitoes, simulids, and culicoids, with their epidemiology dependent on the geographical distribution of competent vectors. Eight species of Dirofilaria have been reported so far in Brazil, of which six parasitize non-human primates, deer, procyonids, and marsupials. Here, we investigated the occurrence of Onchocercidae in wild felids (i.e., Panthera onca, Puma concolor, Herpailurus yagouaroundi, Leopardus geoffroyi, Leopardus guttulus, Leopardus pardalis, Leopardus wiedii, Leopardus munoai) from different locations in Brazil. Overall, 82 samples (n = 63 blood; n = 19 tissues) were molecularly screened for cytochrome c oxidase subunit-1 (cox1) gene. Four (i.e., 4.8%) wild felid samples were positive, and at BLAST analysis, the obtained sequences showed varying percentage of nucleotide identity with the genera Brugia (i.e., 87-88%), Setaria (i.e., 89%), and D. immitis (i.e., 94.4%). Phylogenetic analyses clustered sequences obtained into three distinct clades, one with D. immitis and the remaining two with other Onchocercidae spp. Data herein obtained highlight the need for a more comprehensive understanding of the diversity and biology of Onchocercidae in South America in order to assess the potential impact that these species may have for domestic and wild animals, as well as humans.
Assuntos
Dirofilaria immitis , Dirofilariose , Felidae , Animais , Brasil/epidemiologia , Felidae/parasitologia , Dirofilariose/parasitologia , Dirofilariose/epidemiologia , Dirofilaria immitis/genética , Dirofilaria immitis/isolamento & purificação , Dirofilaria immitis/classificação , Filogenia , Complexo IV da Cadeia de Transporte de Elétrons/genética , Animais Selvagens/parasitologia , Análise de Sequência de DNA , DNA de Helmintos/genética , Dados de Sequência MolecularRESUMO
Rumen flukes cause heavy economic losses in the ruminant industry worldwide, especially in tropical and subtropical countries. This study estimated the prevalence of rumen flukes in buffaloes, identified the species diversity, and determined risk factors associated with rumen fluke prevalence in Perak, Peninsular Malaysia. A cross-sectional study was conducted, and 321 faecal samples were collected from six buffalo farms. A structured questionnaire was developed, and farmers were interviewed to obtain information regarding risk factors associated with rumen fluke infection. The faecal samples were examined using sedimentation and Flukefinder® techniques. Genomic DNA was extracted from the fluke eggs recovered using the Flukefinder® method, and the internal transcribed spacer 2 (ITS2) fragment was amplified and sequenced to facilitate species identification. The results showed that the overall prevalence of rumen fluke across the sampled farms was 40.2% (129/321). Three rumen fluke species were identified, namely, Fischoederius elongatus, F. cobboldi, and Orthocoelium streptocoelium. Several management factors had a significant association (P < 0.05) with rumen fluke prevalence, including production type, cleaning of the stable, drinking water system, flooding around the farm, grazing system, pasture sharing with other livestock, and deworming program. This work constitutes the first attempt to understand the epidemiology of rumen fluke infection in the region and suggests that good farm management, pasture management, choosing appropriate drugs, and proper husbandry practices may improve buffalo health and production in areas where rumen flukes are prevalent.
Assuntos
Búfalos , Fazendas , Fezes , Rúmen , Infecções por Trematódeos , Animais , Búfalos/parasitologia , Malásia/epidemiologia , Prevalência , Estudos Transversais , Fatores de Risco , Rúmen/parasitologia , Fezes/parasitologia , Infecções por Trematódeos/epidemiologia , Infecções por Trematódeos/veterinária , Infecções por Trematódeos/parasitologia , DNA de Helmintos/genética , Inquéritos e QuestionáriosRESUMO
Echinococcosis is a worldwide disease endemic to the western region of China. In 2023, echinococcosis was detected in one of 27 wild boars (Sus scrofa) in Yili Prefecture, Xinjiang, northwestern China. Histopathological staining and full sequence mitochondrial (mt) analysis were used to determine the infection genotype. Echinococcus granulosus was detected in the wild boar liver, and the cystic lesion characteristics indicated the E. granulosus genotype (G1). This case is the first confirmation of wild boar serving as a transmitter for the G1 genotype of E. granulosus within China. These findings suggest that surveillance is needed to assess the risk of E. granulosus sensu lato transmission to humans and wild animals.
Assuntos
Equinococose , Echinococcus granulosus , Genótipo , Sus scrofa , Doenças dos Suínos , Animais , China , Echinococcus granulosus/genética , Echinococcus granulosus/isolamento & purificação , Echinococcus granulosus/classificação , Sus scrofa/parasitologia , Doenças dos Suínos/parasitologia , Suínos , Equinococose/veterinária , Equinococose/parasitologia , Equinococose/epidemiologia , Fígado/parasitologia , Fígado/patologia , Análise de Sequência de DNA , DNA Mitocondrial/genética , DNA de Helmintos/genética , FilogeniaRESUMO
This study represents the first investigation into the occurrence and identification of Metastrongylus spp. in wild boars (Sus scrofa) in Iran, utilizing both molecular and morphological methods. Thirteen wild boars from Kerman Province were examined, with 92.3% found to be infected with at least one species of Metastrongylus. Mixed infections were observed in 38.46% of the animals. Morphological and molecular analyses confirmed the presence of M. pudendotectus and M. salmi, with prevalence rates of 76.9% and 53.9%, respectively. Histopathological examination revealed transverse and longitudinal sections of Metastrongylus parasites within the airways, causing partial to complete obstruction, interstitial pneumonia, and inflammatory responses. The study also highlights the public health significance of these parasites. The higher prevalence observed compared to earlier studies suggests changes in environmental conditions, host dynamics, or agricultural practices as possible factors, warranting further investigation. The findings underscore the need for comprehensive surveillance and control measures to mitigate the risk of zoonotic transmission, particularly in regions with significant wild and domestic swine populations. This study contributes to the understanding of Metastrongylus spp. distribution and their pathological impact, emphasizing the ecological importance of wild boars and the necessity for continued monitoring and research to prevent and control infections in both animal and human populations.
Assuntos
Metastrongyloidea , Infecções por Strongylida , Sus scrofa , Doenças dos Suínos , Animais , Irã (Geográfico)/epidemiologia , Infecções por Strongylida/veterinária , Infecções por Strongylida/parasitologia , Infecções por Strongylida/epidemiologia , Sus scrofa/parasitologia , Doenças dos Suínos/parasitologia , Doenças dos Suínos/epidemiologia , Suínos , Metastrongyloidea/isolamento & purificação , Metastrongyloidea/classificação , Metastrongyloidea/genética , Prevalência , Pulmão/parasitologia , DNA de Helmintos/genética , Masculino , Análise de Sequência de DNA , Coinfecção/parasitologia , Coinfecção/veterinária , Coinfecção/epidemiologiaRESUMO
Diphyllobothriosis, a fish-borne zoonosis in South America, is mainly caused by the Pacific broad tapeworm Adenocephalus pacificus Nybelin, 1931, a parasite of considerable concern in fishery resources due to its impact on public health. A new diphyllobothrid, Diphyllobothrium sprakeri Hernández-Orts et al. Parasites Vectors 14:219, 2021, was recently described from sea lions from the Pacific Coast, but marine fish acting as intermediate hosts are unknown. The objective of this study was to confirm the presence of plerocercoid larvae of Diphyllobothriidae Lühe, 1910 (Cestoda: Diphyllobothriidea) in nine fish species of commercial importance in Peru. Of a total of 6999 fish (5861 Engraulis ringens, 853 Sciaena deliciosa, 6 Sciaena callaensis, 171 Scomber japonicus, 40 Trachurus murphyi, 40 Ariopsis seemanni, 18 Merluccius peruanus, 5 Sarda chiliensis, and 5 Coryphaena hippurus), 183 were infected with plerocercoid larvae, representing a total prevalence of 2.61% and a mean intensity of 3.2. Based on mtDNA cox1 sequences of 43 plerocercoids, a phylogenetic analysis revealed that 41 belong to A. pacificus and two to D. sprakeri. These findings are first molecular data for D. sprakeri larvae, and the infections of E. ringens and T. murphyi by plerocercoid larvae represent the first records of intermediate/paratenic hosts for this species. Hence, the findings of the current study enhance our understanding of the presence of diphyllobothriid species in commercial fish from the Southeastern Pacific Ocean and their potential impact on seafood safety for local human populations.
Assuntos
Doenças dos Peixes , Peixes , Larva , Animais , Peru/epidemiologia , Doenças dos Peixes/parasitologia , Doenças dos Peixes/epidemiologia , Peixes/parasitologia , Prevalência , Larva/classificação , Larva/crescimento & desenvolvimento , Larva/genética , Filogenia , Infecções por Cestoides/veterinária , Infecções por Cestoides/parasitologia , Infecções por Cestoides/epidemiologia , Cestoides/genética , Cestoides/classificação , Cestoides/isolamento & purificação , Diphyllobothrium/genética , Diphyllobothrium/classificação , Diphyllobothrium/isolamento & purificação , Difilobotríase/epidemiologia , Difilobotríase/parasitologia , Difilobotríase/veterinária , DNA de Helmintos/genéticaRESUMO
During nematode surveys of natural vegetation in forests of La Cima de Copey de Dota, San José, San José province, Costa Rica, a Xenocriconemella species closely resembling X. macrodora and related species was found. Integrative taxonomical approaches demonstrated that it is a new species described herein as X. costaricense sp. nov. The new species is parthenogenetic (only females have been detected) and characterised by a short body (276-404 µm); lip region with two annuli, not offset, not separated from body contour; first lip annulus partially covering the second lip annulus. Stylet thin, very long (113-133 µm) and flexible, occupying 30.5-47.8% of body length. Excretory pore located from one or two annuli anterior to one or two annuli posterior to level of stylet knobs, at 42 (37-45) µm from anterior end. Female genital tract monodelphic, prodelphic, outstretched, and occupying 35-45% of body length, with vagina slightly ventrally curved (14-18 µm long). Anus located 6-11 annuli from the tail terminus. Tail conoid and bluntly rounded terminus, the last 2-3 annuli oriented dorsally. Results of molecular characterisation and phylogenetic analyses of D2-D3 expansion segments of 28S rRNA, ITS, and partial 18S rRNA, as well as cytochrome oxidase c subunit 1 gene sequences further characterised the new species and clearly separated it from X. macrodora and other related species (X. iberica, X. paraiberica, and X. pradense).
Assuntos
Filogenia , Animais , Costa Rica , Feminino , Masculino , Nematoides/classificação , Nematoides/anatomia & histologia , Nematoides/genética , DNA Ribossômico/genética , RNA Ribossômico 28S/genética , DNA de Helmintos/genética , Florestas , Análise de Sequência de DNARESUMO
New morphological and molecular data were generated for trematodes recovered from the intestines of the fish Pseudaspius hakonensis from two locations in the south of the Russian Far East. Morphologically, these trematodes are identical to Pseudozoogonoides ugui (Microphalloidea: Zoogonidae) from Japan. According to results of phylogenetic analysis based on 28S rDNA sequence data, P. ugui was closely related to Zoogonoides viviparus, and P. subaequiporus appears as a sister taxon to these two species. Genetic distance values, calculated based on both 28S rDNA and ITS2 rDNA, between P. ugui and Z. viviparus represents an interspecific differentiation level. Our results have an ambiguous explanation, indicating that the implication of the presence of one or two compact vitellarial aggregations for the differentiation of Zoogonoides and Pseudozoogonoides should be reconsidered or that our results open up the question of the taxonomical status of trematodes previously denoted as Z. viviparus and P. subaequiporus.
Assuntos
DNA de Helmintos , DNA Ribossômico , Doenças dos Peixes , Filogenia , RNA Ribossômico 28S , Trematódeos , Infecções por Trematódeos , Animais , Trematódeos/genética , Trematódeos/classificação , Trematódeos/anatomia & histologia , RNA Ribossômico 28S/genética , Doenças dos Peixes/parasitologia , Infecções por Trematódeos/parasitologia , Infecções por Trematódeos/veterinária , DNA de Helmintos/genética , DNA Ribossômico/genética , DNA Espaçador Ribossômico/genética , Federação Russa , Análise de Sequência de DNA , Intestinos/parasitologiaRESUMO
The Cyathocotylidae Mühling, 1898 is a family of primitive diplostomoid trematodes important for understanding the evolution of the superfamily Diplostomoidea. However, cyathocotylids remain poorly studied with the use of molecular techniques. In this study we sequenced the 5.8S + ITS2 region, 28S rRNA, and cox1 genes of two cyathocotylid species and obtained new morphological data on them. We propose Georduboisia nom. nov. instead of the preoccupied name Duboisia Szidat, 1936 (junior homonym of Duboisia Stremme, 1911). Adults of Georduboisia cf. teganuma (Ishii, 1935) and Paracoenogonimus ovatus Katsurada, 1914 were collected from fish-eating birds in the south of the European part of Russia. Georduboisia cf. teganuma was very similar to G.teganuma but differed from it in the shape of the testes. The 28S rRNA gene dataset provided the best-resolved phylogeny of the Cyathocotylidae to date. In the phylogram based on partial sequences of this gene, P. ovatus was close to members of Holostephanoides Dubois, 1983, Neogogatea Chandler & Rausch, 1947 and Gogatea Szidat, 1936. Georduboisia cf. teganuma clustered with members of Cyathocotyle Mühling, 1896 and Holostephanus Szidat, 1936. Phylogenetic analysis based on the 5.8S + ITS2 dataset showed that adults of P. ovatus examined in our study were conspecific with the metacercariae from the musculature of fish collected in Hungary and Italy. It also revealed probable misidentifications of larvae and adults of cyathocotylids whose sequences are deposited in GenBank NCBI.
Assuntos
DNA de Helmintos , Filogenia , RNA Ribossômico 28S , Trematódeos , Animais , Trematódeos/classificação , Trematódeos/genética , Trematódeos/anatomia & histologia , Trematódeos/isolamento & purificação , RNA Ribossômico 28S/genética , DNA de Helmintos/genética , Federação Russa , Aves/parasitologia , DNA Espaçador Ribossômico/genética , Análise de Sequência de DNA , Infecções por Trematódeos/parasitologia , Infecções por Trematódeos/veterinária , RNA Ribossômico 5,8S/genética , Doenças das Aves/parasitologiaRESUMO
The deepest recorded depth for trematodes currently stands at approximately 6200 m. This depth record was achieved solely through sequence datasets of Lepidapedon sp. obtained from a gastropod. Given that trematodes of this genus typically use fish as definitive hosts, the origin of the trematode sequence was thought to be larval stages. However, the specific species remained unclear owing to the absence of reported adult-stage sequences. In the present study, we definitively identified the deepest trematode as Lepidapedon oregonense by comparing 28S ribosomal DNA sequences from adult worms from the macrourid fish Coelorinchus gilberti with data from the gastropod in the previous study.
Assuntos
DNA de Helmintos , DNA Ribossômico , Filogenia , RNA Ribossômico 28S , Trematódeos , Animais , Trematódeos/classificação , Trematódeos/genética , Trematódeos/isolamento & purificação , RNA Ribossômico 28S/genética , DNA de Helmintos/genética , DNA Ribossômico/genética , Gastrópodes/parasitologia , Análise de Sequência de DNA , Peixes/parasitologia , Doenças dos Peixes/parasitologia , Infecções por Trematódeos/parasitologia , Infecções por Trematódeos/veterináriaRESUMO
The genus Ancyrocephalus sensu lato is a large assemblage of species of dactylogyrid monopisthocotyleans without clear taxonomic boundaries. Despite an urgent need for revision, only three representatives of this taxon have been molecularly characterised so far. We found specimens of Ancyrocephalus curtus, a previously non-genotyped species, in gills of Perccottus glenii caught in the River Syumnyur, Amur Basin, Russia. The aim of this study was to assess the phylogenetic position of this parasite using partial sequences of 28S rRNA gene. In the phylogenetic tree, A. curtus appeared as a sister taxon to the dactylogyrine genus Gobioecetes. The new molecular evidence supports the hypothesis about the non-monophyletic status of Ancyrocephalus sensu lato.
Assuntos
Doenças dos Peixes , Brânquias , Perciformes , Filogenia , RNA Ribossômico 28S , Animais , Doenças dos Peixes/parasitologia , Brânquias/parasitologia , Perciformes/parasitologia , RNA Ribossômico 28S/genética , Federação Russa , Rios/parasitologia , Infecções por Trematódeos/parasitologia , Infecções por Trematódeos/veterinária , Platelmintos/classificação , Platelmintos/genética , Platelmintos/isolamento & purificação , DNA de Helmintos/genética , Trematódeos/genética , Trematódeos/classificação , Trematódeos/isolamento & purificação , DNA Ribossômico/genética , Análise de Sequência de DNARESUMO
As part of a parasitological survey, several specimens of two new monopisthocotylean species, Neotetraonchus celsomanueli sp. nov. and N.peruvianus sp. nov. (Dactylogyridea, Dactylogyridae), were collected from the gill filaments of the Peruvian sea catfish Galeichthys peruvianus (Siluriformes, Ariidae) off Puerto Pizarro, Tumbes region, Peru. Neotetraonchus celsomanueli sp. nov. is characterised by an MCO with a T-shaped distal end and an accessory piece that is ribbed and expanded proximally with a worm-shaped termination. Neotetraonchus peruvianus sp. nov. is typified by its MCO, which has a sledgehammer-shaped distal end and an accessory piece with a claw-shaped distal end. Additionally, N.peruvianus sp. nov. is characterised by its jellyfish-shaped onchium. A partial 28S rDNA sequence was obtained from N.celsomanueli sp. nov., and a phylogenetic analysis was conducted. This analysis revealed the phylogenetic position of Neotetraonchus celsomanueli sp. nov. within a clade comprising monopisthocotylean parasites of diadromous and marine ariid catfishes, including Hamatopeduncularia spp., Chauhanellus spp., Thysanotohaptor Kritsky, Shameem, Kumari & Krishnaveni, , and Neocalceostomoides spinivaginalis Lim, 1995. This finding brings the number of known Neotetraonchus species to seven and represents the first described Neotetraonchus species infecting marine catfishes from Peru.
Assuntos
Peixes-Gato , Doenças dos Peixes , Brânquias , Filogenia , Animais , Peixes-Gato/parasitologia , Peru , Doenças dos Peixes/parasitologia , Brânquias/parasitologia , Infecções por Trematódeos/veterinária , Infecções por Trematódeos/parasitologia , DNA Ribossômico/genética , Trematódeos/classificação , Trematódeos/genética , Trematódeos/anatomia & histologia , Trematódeos/isolamento & purificação , DNA de Helmintos/genética , RNA Ribossômico 28S/genética , Platelmintos/classificação , Platelmintos/genética , Platelmintos/anatomia & histologia , Platelmintos/isolamento & purificação , Análise de Sequência de DNARESUMO
The aim of this study was to characterize the Tunisian Fasciola spp. flukes by morphometric and molecular analyses. Flukes were collected from livers of sheep slaughtered in Sejnane slaughterhouses (Bizerte gouvernorate, Northwest Tunisia) between January and March 2021.Five morphometric parameters were determined for all the liver flukes, as follows: (i) total body length (BL), (ii) distance between ventral sucker and the tail (VS-T), (iii) distance between oral sucker and ventral sucker (OS-VS), (iv) abdomen diameter (AD), (v) tail diameter (TD) and the body length to width ratio (BL/BW). Molecular identification of the fluke specimens was carried out by polymerase chain reaction, restriction fragment polymorphism (PCR-RFLP) of a 680 bp sequence of the internal transcribes spacer 1 (ITS1) gene and by amplification, sequencing, and phylogenetic analysis of a 500 bp sequence of the ITS2 gene. Morphometric measurements showed that the mean of the total body length of the adult flukes was 21.1 ± 2.7 mm with minimum and maximum lengths of 13 and 31 mm, respectively. The PCR-RFLP analysis revealed a single profile consisting of three bands of approximately 370, 100, and 60 bp. Fasciola sequences described in the present study (GenBank numbers: OQ457027 and OQ457028) showed 99.58-100% identity to Fasciola hepatica. In conclusion, the results of this study show that molecular and phylogenetic analyses confirm the presence of a single species of F. hepatica in the Sejnane region Northwest of Tunisia. However, further studies are needed to identify the occurrence of Fasciola species in other Tunisian regions.
Assuntos
Doenças dos Bovinos , Fasciola hepatica , Fasciola , Fasciolíase , Ovinos/genética , Animais , Bovinos , Fasciola/genética , Filogenia , Tunísia/epidemiologia , Fasciolíase/epidemiologia , Fasciolíase/veterinária , Fasciola hepatica/genética , Doenças dos Bovinos/epidemiologia , DNA de Helmintos/genéticaRESUMO
Loop-mediated isothermal amplification (LAMP) is the most popular technology for point-of-care testing applications due its rapid, sensitive and specific detection with simple instrumentation compared to PCR-based methods. Many systems for reading the results of LAMP amplifications exist, including real-time fluorescence detection using fluorophore-labelled probes attached to oligonucleotide sequences complementary to the target nucleic acid. This methodology allows the simultaneous detection of multiple targets (multiplexing) in one LAMP assay. A method for multiplexing LAMP is the amplification by release of quenching (DARQ) technique by using a 5'-quencher modified LAMP primer annealed to 3'-fluorophore-labelled acting as detection oligonucleotide. The main application of multiplex LAMP is the rapid and accurate diagnosis of infectious diseases, allowing differentiation of co-infecting pathogens in a single reaction. Schistosomiasis, caused among other species by Schistosoma mansoni and strongyloidiasis, caused by Strongyloides stercoralis, are the most common helminth-parasite infections worldwide with overlapping distribution areas and high possibility of coinfections in the human population. It would be of great interest to develop a duplex LAMP to detect both pathogens in the same reaction. In this study, we investigate the use of our two previously developed and well-stablished LAMP assays for S. mansoni and Strongyloides spp. DNA detection in a new duplex real-time eight-primer system based on a modified DARQ probe method that can be performed in a portable isothermal fluorimeter with minimal laboratory resources. We also applied a strategy to stabilize the duplexed DARQ-LAMP mixtures at room temperature for use as ready-to-use formats facilitating analysis in field settings as point-of-care diagnostics for schistosomiasis and strongyloidiasis.
Assuntos
Esquistossomose , Strongyloides stercoralis , Estrongiloidíase , Animais , Humanos , Schistosoma mansoni/genética , Sistemas Automatizados de Assistência Junto ao Leito , DNA de Helmintos/genética , Técnicas de Amplificação de Ácido Nucleico/métodos , Strongyloides stercoralis/genética , Oligonucleotídeos , Corantes Fluorescentes , Sensibilidade e EspecificidadeRESUMO
Members of the genus Strigea Abildgaard, 1790 are endoparasites of birds distributed worldwide. Adults of an undescribed species of the genus Strigea were collected from the intestines of two hawk species (Rupornis magnirostris and Accipiter coperii). Other species identified as Parastrigea macrobursa that were described in Argentina were also recovered from two hawk species (Buteogallus urubitinga and Buteogallus anthracinus) in three localities along the coasts of Mexico. Specimens of the two species were sequenced for three molecular markers, the internal transcribed spacers locus (ITS1-5.8S rDNA- ITS2) and the domains D1-D3 from the large subunit from nuclear ribosomal DNA and the cytochrome c oxidase subunit 1 from mitochondrial DNA. The newly sequenced specimens were aligned with other strigeids sequences downloaded from GenBank. Maximum likelihood and Bayesian analyses inferred with each molecular marker revealed that our specimens of Strigea sp. formed an independent lineage, which is recognized herein as a new species, Strigea magnirostris n. sp., representing the first species in Mexico and the 16th in the Neotropical region. Morphologically, the new species is distinguished from other congeneric species from the Americas by having an oral sucker with several papillae around it, well-developed pseudosuckers (118-248 µm), a tegument covered with tiny spines, a larger cone genital (193-361 × 296-637) and a larger copulatory bursa (247-531 × 468-784). Our phylogenetic analyses revealed that P. macrobursa is not closely related to other members of the genus Parastrigea and is nested within Strigea, suggesting that P. macrobursa should be transferred to Strigea to form Strigea macrobursa n. comb., expanding its distribution range from Mexico to Argentina. Finally, the analyses also revealed that the taxonomy and systematics of Strigea should be re-evaluated, combining morphological and molecular characteristics.
Assuntos
Aves Predatórias , Trematódeos , Animais , Filogenia , Teorema de Bayes , Aves , DNA Ribossômico/genética , México , DNA de Helmintos/genéticaRESUMO
Scaphanocephalus is a small trematode genus belonging to the family Opistorchiidae. The genus currently contains only three species associated with marine fish as intermediate hosts and fish-eating birds as definitive hosts. Here, specimens of Scaphanocephalus were collected from the Osprey, Pandion haliaetus, and the White mullet, Mugil curema in the Yucatán Peninsula, Mexico. We report for the first-time DNA sequences of adult specimens of Scaphanocephalus, particularly S. expansus, as well as a sequence of a different species sampled as metacercaria. Morphological comparisons of Scaphanocephalus expansus confirmed the identity of the adult specimens, with minor morphological variations; Scanning electron photomicrographs were included, and the species was re-described. Phylogenetic analysis based on 28S rDNA sequences showed that Scaphanocephalus is monophyletic within Opisthorchiidae and consists of three independent lineages. Sequences of adults are identical to those of S. expansus. Instead, the sequence of the metacercaria sampled from the mesentery of Mugil curema nested with specimens reported as Scaphanocephalus sp. from a labrid fish in the Mediterranean Sea, herein named it as Scaphanocephalus sp. 2.