ABSTRACT
OBJECTIVE: To investigate the origin of Biomphalaria straminea in China, so as to provide insights into assessment of schistosomiasis mansoni transmission risk and B. straminea control. METHODS: Guanlan River, Dasha River, Shenzhen Reservoir, upper and lower reaches of Kuiyong River, and Xinzhen River in Shenzhen, China, were selected as sampling sites. Ten Biomphalaria samples were collected from each site, and genomic DNA was extracted from Biomphalaria samples. DNA samples were obtained from 15 B. straminea sampled from 5 sampling sites in Minas Gerais State, Pará State, Federal District, Pernambuco State, and Sao Paulo State in Brazil, South America. Cytochrome c oxidase I (COI) and mitochondrial 16S ribosomal RNA (16S rRNA) genes were sampled using the above DNA templates, and the amplified products were sequenced. The COI and 16S rRNA gene sequences were downloaded from GenBank, and the sampling sites were acquired. All COI and 16S rRNA gene sequences were aligned and evolutionary trees of B. straminea were created based on COI and 16S rRNA gene sequences to identify the genetic similarity and evolutionary relationship between B. straminea samples from China and South America. RESULTS: A total of 60 COI gene sequences with a length of 529 bp and 3 haplotypes were obtained from B. straminea sampled from China. There were 165 COI gene sequences of B. straminea retrieved from GenBank, and following alignment with the above 60 gene sequences, a total of 33 haplotypes were obtained. Phylogenetic analysis showed that the three haplotypes of B. straminea from China were clustered into one clade, among which the haplotype China11 and three B. straminea samples from Brazil retrieved from GenBank belonged to the same haplotype. Geographical evolution analysis showed that the B. straminea samples from three sampling sites along eastern coasts of Brazil had the same haplotype with China11, and B. straminea samples from other two sampling sites were closely, genetically related to China11. A total of 60 16S rDNA gene sequences with approximately 322 bp in length were amplified from B. straminea in China, with 2 haplotypes identified. A total of 70 16S rDNA gene sequences of B. straminea were captured from GenBank. Phylogenetic analysis showed that Biomphalaria snails collected from China were clustered into a clade, and the haplotype China64 and the haplotype 229BS from Brazil shared the same haplotype. The 49 16S rDNA gene sequences of B. straminea from 25 sampling sites in southern Brazil, which were captured from GenBank, were included in the present analysis, and the B. straminea from 3 sampling sites shared the same haplotype with China64 in China. Geographical evolution analysis based on COI and 16S rRNA gene sequences showed that B. straminea sampled from eastern coastal areas of Brazil shared the same haplotypes in two gene fragment sequences with Biomphalaria snails collected from China. CONCLUSIONS: The Biomphalaria snails in China are characterized as B. straminea, which have a low genetic diversity. The Biomphalaria snails in China have a high genetic similarity with B. straminea sampled from eastern coastal areas of Brazil, which may have originated from the eastern coastal areas of Brazil.
Subject(s)
Biomphalaria , Phylogeny , RNA, Ribosomal, 16S , Animals , China , RNA, Ribosomal, 16S/analysis , RNA, Ribosomal, 16S/genetics , Biomphalaria/genetics , Biomphalaria/parasitology , Electron Transport Complex IV/genetics , Electron Transport Complex IV/analysis , HaplotypesABSTRACT
Thelazia callipaeda (Nematoda: Spirurida: Thelaziidae) parasitizes the eyes of dogs, cats, humans, and various wild mammals, and is transmitted by drosophilid flies. In Japan, T. callipaeda is considered an emerging parasite that has expanded its endemic region northward. However, reports of its detection in mammals other than domestic animals and humans are scarce. This study reports the detection of T. callipaeda in Japanese red fox (Vulpes vulpes japonica), masked palm civet (Paguma larvata), Japanese badger (Meles anakuma), Japanese black bear (Ursus thibetanus japonicus), raccoon (Procyon lotor), Japanese raccoon dog (Nyctereutes viverrinus), domestic dog (Canis lupus familiaris), domestic cat (Felis silvestris catus), and human. Of these, the Japanese red fox, masked palm civet, Japanese badger, and Japanese black bear have been reported as novel host records. Sequence analysis of the cytochrome c oxidase subunit I gene of T. callipaeda revealed two unique lineages specific to Japan, with no regional or host species differences. These results suggest a wide host range for T. callipaeda, highlighting the significant role of wildlife as a reservoir for this parasite in Japan.
Subject(s)
Host Specificity , Mustelidae , Spirurida Infections , Thelazioidea , Ursidae , Animals , Thelazioidea/isolation & purification , Thelazioidea/classification , Thelazioidea/genetics , Japan , Spirurida Infections/veterinary , Spirurida Infections/parasitology , Spirurida Infections/epidemiology , Humans , Dogs , Mustelidae/parasitology , Ursidae/parasitology , Cats , Zoonoses/parasitology , Foxes/parasitology , Viverridae/parasitology , Raccoons/parasitology , Electron Transport Complex IV/analysis , Electron Transport Complex IV/genetics , Phylogeny , Animals, Wild/parasitologyABSTRACT
Pacific pink salmon (Oncorhynchus gorbuscha) were deliberately introduced to rivers surrounding the White Sea and has spread to Norway and several other countries surrounding the North Atlantic Ocean. In August 2021, a female pink salmon displaying pale gills and abnormal behaviour was captured in River Lakselva in Northern Norway and later submitted to the Norwegian Veterinary Institute (NVI) for post-mortem examination. Histological examination of organ samples revealed structures indicative of systemic ichthyophoniasis, caused by Ichthyophonus sp. The parasites appeared to be especially abundant in the heart and skeletal musculature, and local tissue responses were assessed to be absent or very mild. Sequences of the ribosomal 18S rRNA and the mitochondrial cytochrome oxidase 1 (CO1) genes confirmed the diagnosis and identified the pathogen as Ichthyophonus sp. The CO1 sequence further established that the isolate from pink salmon was most similar to sequences of Ichthyophonus sp. from Atlantic salmon, Salmo salar, from the Atlantic Ocean on the east coast of the US and from Atlantic herring, Clupea harengus, from Iceland. We here report the first detection of Ichthyophonus sp. in pink salmon in the North Atlantic Ocean.
Subject(s)
Fish Diseases , RNA, Ribosomal, 18S , Animals , Fish Diseases/parasitology , Fish Diseases/diagnosis , Norway , Atlantic Ocean , RNA, Ribosomal, 18S/genetics , Female , Mesomycetozoea/genetics , Mesomycetozoea/isolation & purification , Mesomycetozoea Infections/parasitology , Mesomycetozoea Infections/epidemiology , Salmon/parasitology , Electron Transport Complex IV/genetics , Electron Transport Complex IV/analysis , Phylogeny , Introduced SpeciesABSTRACT
The hemlock woolly adelgid, Adelges tsugae Annand (Hemiptera: Adelgidae), is an invasive pest causing significant ecological and economic damage to certain hemlock tree (Tsuga (Endlicher) Carrière, Pinales:Pinaceae) species. In response to this invasive threat, biological control strategies have been implemented, introducing natural predators such as Laricobius nigrinus Fender (Coleoptera: Derodontidae) and, more recently, Laricobius osakensis Montgomery and Shiyake (Coleoptera: Derodontidae), as specialist predators against A. tsugae. However, the genetic and morphological similarities between L. osakensis and both L. nigrinus and the native beetle, Laricobius rubidus LeConte (Coleoptera: Derodontidae), pose challenges in their identification. Effective monitoring of released predators is integral to evaluating the success of biological control measures. Environmental DNA (eDNA) holds potential for various detection applications, including species monitoring. In this study, we developed specific primers and probes targeting the mitochondrial cytochrome oxidase 1 gene sequences, achieving high specificity despite their 95% sequence similarity. With an optimal annealing temperature of 60 °C, our tools effectively differentiated L. osakensis from the other 2 beetles and demonstrated eDNA detection sensitivity down to 2 copies/µl. This research underscores the potential of precise molecular tools for advancing biological control and biodiversity assessment against invasive threats like A. tsugae.
Subject(s)
Coleoptera , Hemiptera , Pest Control, Biological , Animals , Hemiptera/genetics , DNA, Environmental/analysis , Tsuga , Electron Transport Complex IV/analysis , Electron Transport Complex IV/genetics , Introduced SpeciesABSTRACT
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.
Subject(s)
Fish Diseases , Animals , Egypt/epidemiology , Fish Diseases/parasitology , Fish Diseases/epidemiology , Prevalence , Mediterranean Sea/epidemiology , Female , Male , Ascaridida Infections/veterinary , Ascaridida Infections/parasitology , Ascaridida Infections/epidemiology , Phylogeny , Ascaridoidea/isolation & purification , Ascaridoidea/genetics , Ascaridoidea/classification , Electron Transport Complex IV/analysis , Electron Transport Complex IV/genetics , DNA, Ribosomal Spacer/analysis , DNA, Ribosomal Spacer/genetics , DNA, Helminth/analysisABSTRACT
The box tree moth (BTM), Cydalima perspectalis Walker, is a pest that infests various plants within the Buxus genus. Although a specific parasitoid wasp species associated with the BTM has been observed in the Republic of Korea, no research on this species has been published. Here, we describe the fundamental morphological and biological characteristics of this parasitoid. We have identified the wasp as belonging to the genus Eriborus (Hymenoptera: Ichneumonidae: Campopleginae). Eriborus sp. parasitizes within the living host body, with 1 wasp emerging from each host. The parasitism rate observed in collected BTM populations was 33.1%. The emergence rate was 87.1%, with all emerging adults being females, resulting in a sex ratio of 0. The pupal period avg 9.5 days, and the adult lifespan avg 10.5 days. Eriborus sp. parasitized BTM larvae from the first to the fourth instar and reproduced by thelytokous parthenogenesis. Eriborus sp. exhibited morphological differences compared with previously reported Eriborus species in Korea, particularly in the length of the ovipositor sheath. Additionally, the proportion of the highest similarity in nucleotide sequences of mitochondrial cytochrome oxidase I DNA was only 94.53%, rendering species identification using GenBank's mt cytochrome c oxidase 1 DNA sequences unfeasible. These data suggest that Eriborus sp. could be used as a biological control agent for managing BTM infestations.
Subject(s)
Moths , Wasps , Animals , Wasps/physiology , Moths/parasitology , Female , Male , Republic of Korea , Larva/growth & development , Larva/parasitology , Pupa/parasitology , Pupa/growth & development , Host-Parasite Interactions , Electron Transport Complex IV/analysisABSTRACT
Identifying and analysing distinct blood cells is crucial for the diagnosis and treatment of diseases in the field of biomedicine. The present study was undertaken to study the cytomorphological and cytochemical characteristics of the blood cells of Zoar, a non-descript indigenous breed of chicken extensively reared under backyard poultry farming in Mizoram, India. For this study, 2 mL of blood samples were aseptically collected from the wings veins of 12 chickens and were processed for light microscopic study under standard protocols. The matured erythrocytes were elliptical, while the immature erythrocytes appeared oval. The heterophils were positive for SBB (SBB), Periodic Acid Schiff (PAS), acid phosphatase, alkaline phosphatase and Arylsulphatase while the eosinophils were positive for SBB, PAS, alkaline phosphatase, cytochrome oxidase and peroxidase. The basophils of were positive for toluidine blue while the thrombocytes were positive for PAS. These cytochemical and cytoenzymatic staining properties plays a very important role in diagnosis, differentiation, and classification of leukaemias.
Subject(s)
Chickens , Eosinophils , Erythrocytes , Animals , Chickens/anatomy & histology , India , Erythrocytes/cytology , Eosinophils/cytology , Blood Cells/cytology , Blood Platelets/cytology , Alkaline Phosphatase/blood , Basophils/cytology , Acid Phosphatase/blood , Electron Transport Complex IV/analysisABSTRACT
Spirometra mansoni is a diphyllobothroid cestode and one of the causing agents of sparganosis, a zoonotic foodborne and waterborne infection in humans. This parasite has an indirect life cycle with domestic and wild canids or felids as definitive hosts. The last report of S. mansoni in Costa Rica was done in 2004 by morphological assessment of worms, whereas molecular evidence of this species was obtained recently in the Americas. Herein, we present seven cases of spirometrosis in four dogs, three cats and a coyote from different regions of Costa Rica occurring in a time span of a year. Dog cases presented vomiting, hyporexia, lethargy and diarrhea, whereas cats were mostly asymptomatic. Moreover, the coyote was found with Spirometra sp. proglottids incidentally. Cytochrome oxidase subunit 1 (cox1) sequences of eggs or proglottids derived from all cases were analyzed with a Bayesian Inference phylogenetic tree and a haplotype network. These analyses showed the clustering of S. mansoni from Costa Rica with other sequences derived from Asia and America. Moreover, cox1 sequences clustered in two separate haplotypes, suggesting the high genetic diversity of the species. The present cases represent the first molecular evidence of the parasite in Central America; thus, extending its known range in the American continent.
Subject(s)
Animals, Wild , Cat Diseases , Dog Diseases , Phylogeny , Spirometra , Animals , Cats/parasitology , Dogs , Female , Male , Animals, Wild/parasitology , Cat Diseases/parasitology , Cat Diseases/epidemiology , Cestode Infections/veterinary , Cestode Infections/parasitology , Cestode Infections/epidemiology , Costa Rica/epidemiology , Coyotes/parasitology , Dog Diseases/parasitology , Dog Diseases/epidemiology , Electron Transport Complex IV/analysis , Electron Transport Complex IV/genetics , Spirometra/genetics , Spirometra/isolation & purificationABSTRACT
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.
Subject(s)
Animal Migration , Birds , Electron Transport Complex IV , Ixodidae , Phylogeny , Animals , China , Ixodidae/genetics , Ixodidae/classification , Ixodidae/physiology , Electron Transport Complex IV/genetics , Electron Transport Complex IV/analysis , RNA, Ribosomal/genetics , RNA, Ribosomal/analysis , Nymph/growth & development , Nymph/classification , Nymph/genetics , Nymph/physiology , Arthropod Proteins/genetics , Arthropod Proteins/analysis , DNA, Ribosomal Spacer/analysisABSTRACT
In the present study, tissue samples (tongue, esophagus and heart) were investigated from dromedary camels of India for identification and characterization of Sarcocystis spp. using histopathology, PCR and gene sequencing. Genomic DNA extracted from these tissue samples was used for PCR amplification of the cytochrome c oxidase subunit I gene (cox1) of Sarcocystis spp. and the partial sequence of small subunit ribosomal RNA (18S rRNA) gene of the S. cameli. The PCR products were purified, sequenced and analyzed using bioinformatics tools. Based on phylogenetic analysis of the cox1 gene, the sequences of the present study clustered with those of S. cameli, hosted by dromedary camels of Iraq and a close association was observed with S. masoni hosted by dogs and alpacas of China. Until now, there are no 18S rRNA sequences of S. cameli available in GenBank and this is the first study recording 18S rRNA sequences of S. cameli which were grouped with S. masoni from alpaca of China and guanaco and llama of Argentina in phylogenetic analysis. These findings could be useful for further studies on the characterization through molecular epidemiology, genetic diversity and host specificity of S. cameli.
Subject(s)
Camelus , Phylogeny , RNA, Ribosomal, 18S , Sarcocystis , Sarcocystosis , Animals , Sarcocystis/genetics , Sarcocystis/classification , Sarcocystis/isolation & purification , Camelus/parasitology , Sarcocystosis/veterinary , Sarcocystosis/parasitology , Sarcocystosis/epidemiology , RNA, Ribosomal, 18S/genetics , India/epidemiology , Electron Transport Complex IV/genetics , Electron Transport Complex IV/analysisABSTRACT
Amblyomma integrum is a large gooseberry sized longirostrate tick (when fully repleted) found in India and Sri Lanka. In Kerala (India), this tick is commonly found in the forest and its fringe areas frequently infesting deer and hence it is locally known as "maan chellu / maanunny" (deer tick). In the present study, molecular characterisation and phylogenetic analysis of A. integrum collected from the area grazed by the sambar deer (Rusa unicolor) of Kerala, south India was performed using three molecular markers viz., the mitochondrial cytochrome c oxidase subunit 1 (COI), mitochondrial 16S ribosomal RNA, and nuclear 18S ribosomal RNA genes. Cytochrome c oxidase subunit 1 (COI) gene showed better resolving ability for elucidating the evolutionary relationship of A. integrum and identified two distinct clades, viz., A and B. The Tamil Nadu isolates of south India and Marayoor isolate 1 (from Idukki district of Kerala bordering with Tamil Nadu) belonged to clade A. Majority of Wayanad isolates from Kerala, occupied clade B. The intraspecific genetic distance among the A. integrum species ranged from 0.00 to 13.34%. Between clades A and B, the genetic distance observed was 11.49%. The clade B isolates were genetically close to A. geoemydae (GD: 1.22%). Morphological variations between the clades included darker exoskeletal coloration in clade A and distinct differences in the shape of basis capitulum. Further analysis using Assemble Species by Automatic Partitioning (ASAP) and Generalized Mixed Yule Coalescent (GMYC) provided additional insights. Assemble Species by Automatic Partitioning (ASAP) identified 26 Molecular Operational Taxonomic Units (MOTUs) at a threshold distance of 5.38%, supporting the species partition of A. integrum clade B. Generalized Mixed Yule Coalescent (GMYC) analysis retained the same species complex (A. integrum-geoemydae Complex) inferred from the ASAP analyses. It could be inferred from the present study that the A. integrum clades A and B could be two different putative pseudocryptic species.
Subject(s)
Amblyomma , Phylogeny , RNA, Ribosomal, 16S , RNA, Ribosomal, 18S , Animals , India , RNA, Ribosomal, 18S/analysis , RNA, Ribosomal, 18S/genetics , RNA, Ribosomal, 16S/analysis , RNA, Ribosomal, 16S/genetics , Electron Transport Complex IV/analysis , Electron Transport Complex IV/genetics , Tick Infestations/veterinary , Tick Infestations/parasitology , Tick Infestations/epidemiology , Deer/parasitologyABSTRACT
Mosquito-borne diseases can pose significant burdens. In many countries, they pose a risk to national economies and the well-being of humans and animals. To mitigate this, mosquito surveillance is crucial to assess the real and potential transmission of mosquito-borne diseases. Between 2020 and 2023, mosquito larvae were collected from both indoor and outdoor breeding sites in urban and rural areas of 4 municipalities of Santiago and Boavista Islands in Cabo Verde. Mosquitoes were identified morphologically and by polymerase chain reaction-based techniques that targeted the mitochondrial cytochrome C oxidase subunit I sequence. During this period, 6,825 breeding sites were assessed, and of 8,094 mosquito specimens screened, 194 specimens of Culex thalassius were identified for the first time in the country in 4 municipalities of Santiago and Boavista Islands. This new finding highlights the importance of including entomological surveillance in health systems. Although this species has only been detected on a few islands, it is important to continuously monitor it to determine its distribution, spread/dispersal, density, and potential involvement in pathogen transmission.
Subject(s)
Animal Distribution , Culex , Larva , Animals , Larva/growth & development , Larva/classification , Cabo Verde , Mosquito Vectors/genetics , Electron Transport Complex IV/analysis , Electron Transport Complex IV/geneticsABSTRACT
The aim of the present study was to assess morphologic and genetic data on ascariasis in swine (Sus scrofa domesticus) and humans in low-resource rural and periurban communities in the state of Piauí, Brazil. Our cross-sectional survey included 100 fecal samples obtained from swine and 682 samples from humans. Fifteen pigs were necropsied. Human and porcine fecal samples were examined to identify Ascaris eggs. Parasites obtained in the swine necropsies were studied using scanning electron microscopy (SEM), and the mitochondrial gene encoding the cytochrome oxidase 1 (cox1) enzyme was partially amplified and sequenced for molecular taxonomy and phylogenetic analyses. The overall prevalence of Ascaris eggs in the swine fecal samples was 16/100 (16%). No Ascaris eggs were identified in the human fecal samples. SEM of six worms recovered from pigs demonstrated morphological characteristics of A. suum. Cox1 sequences were compatible with A. suum reference sequences. Original and reference (GenBank) nucleotide sequences were organized into clusters that did not segregate the parasites by host species or and region. The largest haplogroups were dominated by haplotypes H01, H02 and H31. In the communities studied, there was no epidemiological evidence of the zoonotic transmission of ascariasis at the human-swine interface.(AU)
O presente estudo teve como objetivo acessar dados morfológicos e genéticos sobre a ascaridíase em suínos (Sus scrofa domesticus) e humanos, em comunidades rurais e periurbanas no estado do Piauí. O estudo transversal incluiu 100 amostras fecais de suínos e 682 amostras obtidas de humanos. Quinze suínos foram necropsiados. Amostras fecais suínas e humanas foram examinadas para detecção de ovos de Ascaris. Os parasitas adultos, obtidos nas necropsias, foram estudados através de microscopia eletrônica de varredura (MEV), e o gene mitocondrial codificante da enzima citocromo oxidase 1 (cox1) foi parcialmente amplificado e sequenciado para análises filogenéticas e de taxonomia molecular. A prevalência de Ascaris em amostras fecais de suínos foi 16/100 (16%), não sendo identificado nenhum caso de infecção por este parasita em humanos. A análise por MEV de parasitas recuperados de suínos demonstrou características morfológicas de Ascaris suum. As sequências nucleotídicas de cox1 foram compatíveis com A. suum. As sequências originais e de referência (obtidas no GeneBank) foram organizadas em clusters que não segregaram os parasitas por hospedeiro ou região geográfica. Os maiores haplogrupos foram dominados pelos haplótipos H01, H02 e H31. Nas comunidades estudadas, não foi evidenciada transmissão zoonótica de A. suum na interface suíno-humana.(AU)
Subject(s)
Humans , Animals , Ascaridiasis/diagnosis , Swine/genetics , Ascaris suum/genetics , Phylogeny , Brazil , Electron Transport Complex IV/analysisABSTRACT
AIMS: Patients with dermatomyositis (DM) suffer from reduced aerobic metabolism contributing to impaired muscle function, which has been linked to cytochrome c oxidase (COX) deficiency in muscle tissue. This mitochondrial respiratory chain dysfunction is typically seen in perifascicular regions, which also show the most intense inflammatory reaction along with capillary loss and muscle fibre atrophy. The objective of this study was to investigate the pathobiology of the oxidative phosphorylation deficiency in DM. METHODS: Muscle biopsy specimens with perifascicular COX deficiency from five juveniles and seven adults with DM were investigated. We combined immunohistochemical analyses of subunits in the respiratory chain including complex I (subunit NDUFB8), complex II (succinate dehydrogenase, subunit SDHB) and complex IV (COX, subunit MTCO1) with in situ hybridisation, next generation deep sequencing and quantitative polymerase chain reaction (PCR). RESULTS: There was a profound deficiency of complexes I and IV in the perifascicular regions with enzyme histochemical COX deficiency, whereas succinate dehydrogenase activity and complex II were preserved. In situ hybridisation of mitochondrial RNA showed depletion of mitochondrial DNA (mtDNA) transcripts in the perifascicular regions. Analysis of mtDNA by next generation deep sequencing and quantitative PCR in affected muscle regions showed an overall reduction of mtDNA copy number particularly in the perifascicular regions. CONCLUSION: The respiratory chain dysfunction in DM muscle is associated with mtDNA depletion causing deficiency of complexes I and IV, which are partially encoded by mtDNA, whereas complex II, which is entirely encoded by nuclear DNA, is preserved. The depletion of mtDNA indicates a perturbed replication of mtDNA explaining the muscle pathology and the disturbed aerobic metabolism.
Subject(s)
Cytochrome-c Oxidase Deficiency , Dermatomyositis , Adult , Humans , Cytochrome-c Oxidase Deficiency/metabolism , Cytochrome-c Oxidase Deficiency/pathology , Succinate Dehydrogenase/analysis , Succinate Dehydrogenase/metabolism , Dermatomyositis/pathology , Electron Transport , Muscle Fibers, Skeletal/chemistry , Muscle Fibers, Skeletal/metabolism , Muscle Fibers, Skeletal/pathology , DNA, Mitochondrial/genetics , Electron Transport Complex IV/analysis , Electron Transport Complex IV/genetics , Electron Transport Complex IV/metabolism , Muscle, Skeletal/pathologyABSTRACT
Originally described from the masked greenling Hexagrammos octogrammus (Pallas, 1814), the palaeacanthocephalan Echinorhynchus hexagrammi Baeva, 1965 has so far been known from seven species in six families of marine teleosts distributed in the Sea of Okhotsk off Sakhalin and in the Northwestern Pacific off Hokkaido, Japan. In this study, we examined the phylogenetic position of E. hexagrammi based on material obtained from the intestine of an unidentified snailfish, Liparis sp., dredged in Akkeshi Bay, Hokkaido, Japan. We performed an analysis using two gene markers, the mitochondrial cytochrome c oxidase subunit I and the nuclear 28S rRNA, along with other sequences available in public databases. In the resulting tree, E. hexagrammi was more closely related to two species complexes, the E. bothniensis Zdzitowiecki and Valtonen, 1987 complex and the E. gadi Zoega in Müller, 1776 complex, rather than to E. brayi Wayland, Sommerville, and Gibson, 1999, E. cinctulus (Porta, 1905), E. salmonis Müller, 1784, and E. truttae Schrank, 1788. The morphology of the examined material herein identified as E. hexagrammi is briefly described. This study represents the first host record of E. hexagrammi from the snailfish family Liparidae.
Subject(s)
Acanthocephala/anatomy & histology , Acanthocephala/genetics , Fishes/parasitology , Acanthocephala/classification , Animals , Electron Transport Complex IV/analysis , Helminth Proteins , Host-Parasite Interactions , Perciformes/parasitology , RNA, Helminth/analysis , RNA, Ribosomal, 28S/analysisABSTRACT
The diaphragm muscles of 77 free-ranging red deer (Cervus elaphus) were examined for Sarcocystis species in Lithuania. Sarcocysts were detected in 61 out of 77 (79.2%) animals investigated. A total of 60 isolated sarcocysts were identified to species using subunit I of cytochrome c oxidase (cox1) sequence analysis. Overall, seven species, S. entzerothi, S. hjorti, S. iberica, S. linearis, S. pilosa, S. truncata and S. venatoria, were confirmed in Lithuanian red deer. Sarcocystis entzerothi was reported in red deer for the first time. Previously this species was shown to use sika deer as well as roe deer and fallow deer as an intermediate host. Based on cox1, with the addition of the current data, altogether 13 Sarcocystis species have so far been shown to use red deer as an intermediate host. Species detected in red deer demonstrated considerable differences in intraspecific genetic variation at cox1. Genetic distances between different samples of S. hjorti and S. linearis were calculated using principal coordinates analysis (PCoA), implying molecular divergence of same Sarcocystis species using different hosts in the same geographical area and divergence of those employing same intermediate host species from different areas.
Subject(s)
Animal Distribution , Deer , Sarcocystis/isolation & purification , Sarcocystosis/veterinary , Animals , Electron Transport Complex IV/analysis , Helminth Proteins/analysis , Lithuania/epidemiology , Prevalence , Sarcocystis/enzymology , Sarcocystis/genetics , Sarcocystosis/epidemiology , Sarcocystosis/parasitologyABSTRACT
Identification of species involved in cadaveric decomposition, such as scavenger Diptera, is a fundamental step for the use of entomological evidence in court. Identification based on morphology is widely used in forensic cases; however, taxonomic knowledge of scavenger fauna is poor for many groups and for many countries, particularly Neotropical ones. A number of studies have documented the utility of a DNA barcoding strategy to assist in the identification of poorly known and diverse groups, particularly in cases involving immature states or fragmented organisms. To provide baseline knowledge of the diversity of scavenger Diptera in the Valley of Mexico, we generated a DNA barcode collection comprised of sequences of the cytochrome c oxidase subunit 1 (COI) gene for all families sampled at a nature reserve located in this region. We collected and identified specimens on the basis of morphology and a species delimitation analysis. Our analyses of 339 individuals delineated 42 species distributed across nine families of Diptera. The richest families were Calliphoridae (9 species), Sarcophagidae (7 species), and Phoridae (6 species). We found many of the species previously recorded for the Valley of Mexico, plus 18 new records for the region. Our study highlights the utility of DNA barcoding as a first-step strategy to assess species richness of poorly studied scavenger fly taxa.
Subject(s)
Animal Distribution , Calliphoridae/classification , Diptera/classification , Sarcophagidae/classification , Animals , DNA Barcoding, Taxonomic , Electron Transport Complex IV/analysisABSTRACT
Ancylostoma ceylanicum is recognized as the only zoonotic hookworm species that is able to mature into adult stage in the human intestine. While human infections caused by this hookworm species have been reported from neighboring countries and this hookworm is prevalent in dogs in Vietnam, human infection has never been reported in Vietnam. The present study, therefore, aimed to identify human infections with A. ceylanicum in Vietnam. A total of 526 fecal samples from the residents in Long An Province were collected and the presence of hookworm eggs was detected by the Kato-Katz method. The results indicated that the overall prevalence of human hookworm infection was 85/526 (16.2%). After filter paper culture, 3rd stage larvae were successfully obtained from 48 egg-positive samples. The larvae were identified for their species using semi-nested PCR-RLFP on the cox1 gene. As a result, two hookworm species were confirmed; single species infections with Necator americanus or A. ceylanicum, and mixed infections with both species were found in 47.9%, 31.3%, and 20.8% of the samples, respectively.
Subject(s)
Ancylostoma/isolation & purification , Ancylostomiasis/epidemiology , Ancylostomiasis/parasitology , Animals , Electron Transport Complex IV/analysis , Helminth Proteins/analysis , Humans , Prevalence , Vietnam/epidemiologyABSTRACT
The present study describes three new species of monogenean parasites of characid fishes from the Upper Paraná River basin, Brazil: Characithecium paranapanemense n. sp. on Psalidodon paranae and Psalidodon bockmanni, Diaphorocleidus magnus n. sp. on Astyanax lacustris and Psalidodon fasciatus, and Diaphorocleidus neotropicalis n. sp. on Astyanax lacustris and P. bockmanni. An amendment for Diaphorocleidus is proposed, since additional characters observed in the new species required to extend the generic diagnostic features mainly to include: articulation process connecting the base of the MCO with accessory piece present or absent, and accessory piece with variable shapes (plate-like, pincer-shaped, wrench-shaped, sheath-shaped), divided or not into subunits. Characithecium paranapanemense n. sp. can be distinguished from other congeners by the morphology of its MCO and accessory piece. Diaphorocleidus magnus n. sp. differs from most of its congeners by the morphology of its accessory piece, the presence of articulation process connecting the base of the MCO with accessory piece, and the morphology of the sclerotized structures of the haptor. Diaphorocleidus neotropicalis n. sp. can be easily distinguished from its congeners by the morphology of the accessory piece, the sclerotized structures of the haptor and the morphology of the vagina. Molecular data of the new species (partial 28S rDNA and mitochondrial cytochrome oxidase I) were obtained and the first phylogenetic analysis based on 28S rDNA gene sequences for species of Characithecium and Diaphorocleidus are provided. Although Diaphorocleidus and Characithecium share some morphological similarities, phylogenetic analysis indicates that species of these two genera are not closely related.
Subject(s)
Characidae , Fish Diseases/parasitology , Trematoda/classification , Trematode Infections/veterinary , Animals , Brazil/epidemiology , DNA, Helminth/analysis , DNA, Ribosomal/analysis , Electron Transport Complex IV/analysis , Helminth Proteins/analysis , Male , Mitochondrial Proteins/analysis , Prevalence , Trematoda/anatomy & histology , Trematoda/cytology , Trematoda/genetics , Trematode Infections/parasitologyABSTRACT
Current article touched upon the issue of the complicated taxonomic status of some species from the genus Crepidostomum collected from the freshwater fish in the rivers of Primorsky region, Sakhalin, and Hokkaido Islands. Primary morphological analyses showed affiliation of the worms to the species C. farionis (Müller, 1784) Lühe, 1909; C. metoecus Braun, 1900b; C. chaenogobii Yamaguti and Matsumura, 1942; C. nemachilus Krotov, 1959. We described the new species Crepidostomum achmerovi sp. nov. that is a sibling species of C. nemachilus. Molecular-genetic investigation have shown that C. nemachilus and C. achmerovi sp. nov. are closely related to C. metoecus in both 28S rDNA and cox1 mtDNA markers. Crepidostomum nemachilus forms a separate branch within the C. metoecus clade on the 28S BI tree with strong statistical support and separate clade in relation to C. metoecus clade on the cox1 BI tree. Values of p-distances between Crepidostomum species were at intergeneric level. Crepidostomum metoecus species complex including five species (C. metoecus, C. nemachilus, C. oschmarini, C. brinkmanni, and C. achmerovi sp. nov.) was reconsidered as independent genus Crepidostomum sensu stricto. Minimum Spanning Network showed that C. nemachilus, C. metoecus and C. achmerovi sp. nov. were separated by large number of mutational events and represent independent phyletic lines. An amended diagnosis is provided for the subfamily Crepidostomatinae, the genera Crepidostomum s. str. and Stephanophiala Nicoll, 1909, along with keys to species of both genera.