scholarly journals Determination of ribosomal internal transcribed spacer 2 (ITS2) interspecific markers in Fasciola hepatica, Fascioloides magna, Dicrocoelium dendriticum and Paramphistomum cervi (Trematoda), parasites of wild and domestic ruminants

2010 ◽  
Vol 47 (2) ◽  
pp. 76-82 ◽  
Author(s):  
E. Bazsalovicsová ◽  
I. Králová-Hromadová ◽  
M. Špakulová ◽  
M. Reblánová ◽  
K. Oberhauserová
Keyword(s):  
Internal Transcribed Spacer ◽  
Fasciola Hepatica ◽  
Fascioloides Magna ◽  
Internal Transcribed Spacer 2 ◽  
Dicrocoelium Dendriticum ◽  
Domestic Ruminants ◽  
Molecular Differentiation ◽  
Species Specific ◽  
First Time ◽  
Paramphistomum Cervi

AbstractThe species-specific ribosomal internal transcribed spacer 2 (ITS2) markers were designed for PCR-based molecular differentiation of Fasciola hepatica, Fascioloides magna, Dicrocoelium dendriticum and Paramphistomum cervi, liver and stomach flukes of domestic and free living ruminants. Complete ITS2 sequences were obtained for D. dendriticum and P. cervi, for the later species, ITS2 structure was determined for the first time. Intraspecific variation within geographically distant populations was found to be either very low (F. hepatica; D. dendriticum) or even absent (F. magna; P. cervi). ITS2 regions with the absence of intraspecific polymorphisms but with interspecific sequence heterogeneity were applied for design of speciesspecific primers. The specificity of developed primers was tested on genomic DNA isolated from adult individuals of studied fluke species. Application of the primers is of particular value for molecular differentiation of morphologically hardly distinguishable F. hepatica, F. magna and P. cervi eggs after coprological examinations.

Characterization of Dicrocoelium dendriticum haplotypes from sheep and cattle in Iran based on the internal transcribed spacer 2 (ITS-2) and NADH dehydrogenase gene (nad1)

2013 ◽  
Vol 89 (2) ◽  
pp. 158-164 ◽  
Author(s):  
S. Gorjipoor ◽  
M. Moazeni ◽  
H. Sharifiyazdi
Keyword(s):  
Internal Transcribed Spacer ◽  
Nadh Dehydrogenase ◽  
Sequence Data ◽  
Geographic Location ◽  
Nucleotide Substitutions ◽  
Internal Transcribed Spacer 2 ◽  
Dicrocoelium Dendriticum ◽  
Dehydrogenase Gene ◽  
First Time

AbstractThe present study assessed whether the genetic variation among different hosts (sheep and cattle) and geographical isolates (n= 28) of Dicrocoelium dendriticum from Iran is present based on mitochondrial (nad1) and ribosomal (ITS-2) DNA markers. Molecular analysis revealed the presence of at least ten and two distinct haplotypes in the NADH dehydrogenase gene (nad1) and internal transcribed spacer 2 (ITS-2), respectively. The nad1 and ITS-2 sequence data were deposited in GenBank under accession numbers, JX050110–134 and JQ966972–3. According to the results of our study, ND-D and ITS-A are established as being the predominant haplotypes of D. dendriticum in Iran. The Iranian isolates showed a higher intraspecific genetic diversity of 0–0.97% for nad1, compared to 0–0.42% for ITS-2. The alignment and comparison of nad1 and ITS-2 sequences revealed eight and one polymorphic sites, respectively. In the nad1 sequences, six were silent and two nucleotide substitutions were responsible for amino acid alterations. A phylogenetic analysis of the sequence data revealed that host associations and geographic location are not likely useful markers for D. dendriticum haplotype classification. Consequently, sequencing results obtained from the nad1 gene as a mitochondrial marker for the first time in this study would provide a valuable tool to analyse further molecular details of D. dendriticum worldwide.

scholarly journals Molecular discrimination of eggs of cervid trematodes using the Teflon (PTFE) technique for eggshell disruption

2010 ◽  
Vol 47 (3) ◽  
pp. 147-151 ◽  
Author(s):  
K. Oberhauserová ◽  
E. Bazsalovicsová ◽  
I. Králová-Hromadová ◽  
P. Major ◽  
M. Reblánová
Keyword(s):  
Fasciola Hepatica ◽  
Methodological Approach ◽  
Pcr Amplification ◽  
Individual Species ◽  
Its2 Region ◽  
Chemical Reagents ◽  
Domestic Ruminants ◽  
Pcr Products ◽  
Species Specific ◽  
Paramphistomum Cervi

Abstract Molecular comparative analysis of eggs of four liver and stomach flukes of cervids and domestic ruminants, Fasciola hepatica, Fascioloides magna, Dicrocoelium dendriticum and Paramphistomum cervi, was performed using a new methodological approach for eggshell disintegration. Eggs of all species were crushed mechanically by the Teflon method (PTFE) without use of chemical reagents and an efficient disruption of eggshell was checked microscopically. The egg suspension was then subjected to DNA isolation and PCR amplification using species-specific primers that annealed to the internal transcribed spacer 2 (ITS2) region of ribosomal DNA. The size of PCR products of individual species corresponded well to the size of amplicons obtained from adult flukes. The results provided evidence that the Teflon method does not destroy the structure of egg DNA, thus making the procedure broadly applicable during coprological examinations. Molecular markers introduced here are particularly important for blanket screening and differentiation of morphologically hardly distinguishable F. hepatica, F. magna and P. cervi eggs.

Molecular identification of Fascioloides magna (Bassi, 1875) from red deer from South-Western Poland (Lower Silesian Wilderness) on the basis of internal transcribed spacer 2 (ITS-2)

2014 ◽  
Vol 17 (3) ◽  
pp. 523-525 ◽  
Author(s):  
A. M. Pyziel ◽  
A. W. Demiaszkiewicz ◽  
I. Kuligowska
Keyword(s):  
Molecular Analysis ◽  
Cervus Elaphus ◽  
Internal Transcribed Spacer ◽  
Liver Fluke ◽  
Red Deer ◽  
Fasciola Hepatica ◽  
Fallow Deer ◽  
Dama Dama ◽  
Fascioloides Magna ◽  
The Usa

Abstract The study was conducted in 2012-2013 on 75 fecal samples of red deer from the Lower Silesian Wilderness which were examined to determine the prevalence of Fascioloides magna in the game population. Finding liver fluke eggs in a single sample which were larger in size than Fasciola hepatica eggs indicated that further molecular analysis was necessarily. The partial sequence (116 bp long) of ITS-2 of the investigated eggs was identical to the sequences of F. magna from red deer (Cervus elaphus) (GenBank, EF534993; GenBank, EF534992) and from wapiti deer (Cervus elaphus canadensis) (GenBank, EF534994) from Slovakia, as well as from fallow deer (Dama dama) from the USA (GenBank, EF051080). This is the first molecular confirmation of the occurrence of F. magna in Poland.

scholarly journals New data on wild boar (Sus scrofa L.) a dead-end host for large American liver fluke (Fascioloides magna)

2017 ◽  
Vol 54 (1) ◽  
pp. 77-80
Author(s):  
D. Konjević ◽  
M. Bujanić ◽  
V. Erman ◽  
A. Gudan Kurilj ◽  
T. Živičnjak ◽  
...  
Keyword(s):  
Wild Boar ◽  
Sus Scrofa ◽  
Fasciola Hepatica ◽  
Direct Role ◽  
Fascioloides Magna ◽  
Clinical Appearance ◽  
Disease Epidemiology ◽  
Domestic Ruminants ◽  
Dead End ◽  
Host Parasite

Summary Fascioloidosis is a parasitic disease of primary wild and domestic ruminants, caused by a digenean trematode, Fascioloides magna. The final hosts of F. magna are divided according to the host-parasite interactions into definitive, dead end and aberrant. The clinical appearance, pathology, outcome of disease, and its importance in disease epidemiology vary with different host types. According to this division, wild boar (Sus scrofa) are characterized as a dead end host. In this paper we analysed 12 wild boar livers from Croatia. Eleven of them contained pigment traces, pseudocysts, degrading pseudocysts, fluke migratory channels, live and degrading flukes. F. magna eggs were found in pseudocysts, but no eggs were recovered from faeces. Concurrent infection with F. magna and Fasciola hepatica was detected in one liver. According to everything we observed, wild boar currently has no direct role in maintaining and spreading the disease.

Cas de suie de l'érable et de chancre du peuplier dans le canton de Genève

2016 ◽  
Vol 167 (2) ◽  
pp. 98-104
Author(s):  
Bastien Cochard ◽  
François Lefort
Keyword(s):  
Internal Transcribed Spacer ◽  
Pathogenic Fungi ◽  
Fungal Flora ◽  
Acer Campestre ◽  
Cytospora Chrysosperma ◽  
Pure Cultures ◽  
Populus X Euramericana ◽  
Bark Peeling ◽  
Specific Symptoms ◽  
First Time

A case of sooty bark disease and Cytospora poplar canker in the Canton of Geneva In summer 2014, a case of sooty bark disease caused by Cryptostroma corticale on an individual field maple (Acer campestre) and two cases of poplar canker due to Cytospora chrysosperma on Populus x euramericana were identified genetically for the first time on the territory of the Canton of Geneva. In both cases, the trees presented signs of very advanced dieback, accompanied by specific symptoms such as bark peeling and sooty plaques for the maple, and loose twisted bark layers and black colouring of the wood in structural branches of the poplars. Sampling was carried out in the symptomatic areas and components of the fungal flora were isolated in pure cultures in order to identify any pathogenic fungi. The molecular analysis of the rDNA internal transcribed spacer (ITS) sequences made it possible to identify precisely all pure isolates obtained. The results showed a majority presence of C. corticale in the maple tree, and of C. chrysosperma in the two poplars. Both these fungi are little known in Switzerland and Europe, and their presence is maybe associated with changes in the climate.

scholarly journals First Report of Echinococcus ortleppi in Free-Living Wild Boar (Sus scrofa) from Portugal

2021 ◽  
Vol 9 (6) ◽  
pp. 1256
Author(s):  
Teresa Letra Mateus ◽  
Maria João Gargaté ◽  
Anabela Vilares ◽  
Idalina Ferreira ◽  
Manuela Rodrigues ◽  
...  
Keyword(s):  
Wild Boar ◽  
Internal Transcribed Spacer ◽  
Sus Scrofa ◽  
One Health ◽  
Control Strategies ◽  
Free Living ◽  
First Report ◽  
Wildlife Reservoirs ◽  
First Time ◽  
Echinococcus Granulosus Sensu Lato

Cystic echinococcosis (CE) is a zoonosis that is prevalent worldwide. It is considered endemic in Portugal but few studies have been performed on Echinococcus granulosus sensu lato and their hosts. In this study, CE cysts are reported for the first time in a free-living wild boar (Sus scrofa) in Portugal. The presence of the metacestodes in the liver of the wild boar was identified by morphological features, microscopic examination and molecular analysis. The sequencing of part of the DNA nuclear ribosomal internal transcribed spacer-1 (ITS-1) region revealed a G5 genotype that presently corresponds to Echinococcus ortleppi. This is the first report of E. ortleppi in Portugal and to the best of the authors’ knowledge, in Europe. These results suggest that wild boar may be a host of CE, namely, crossing the livestock–wildlife interface, which has important public health implications. Wildlife reservoirs must be taken into account as CE hosts and surveillance of game as well as health education for hunters should be implemented using a One Health approach, with implementation of feasible and tailor-made control strategies, namely, proper elimination of byproducts in the field.

scholarly journals The new Internal Transcribed Spacer 2 diagnostic tool clarifies the taxonomic position and geographic distribution of the North American malaria vector Anopheles punctipennis

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
James M. Hodge ◽  
Andrey A. Yurchenko ◽  
Dmitriy A. Karagodin ◽  
Reem A. Masri ◽  
Ryan C. Smith ◽  
...  
Keyword(s):  
Malaria Vector ◽  
Internal Transcribed Spacer ◽  
Single Species ◽  
Its2 Sequence ◽  
Pathogen Transmission ◽  
Molecular Diagnostic ◽  
Diagnostic Tools ◽  
Sequence Length ◽  
Phylogenetic Position ◽  
Internal Transcribed Spacer 2

Abstract 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.

scholarly journals Bark traits, decomposition and flammability of Australian forest trees

2017 ◽  
Vol 65 (4) ◽  
pp. 327 ◽  
Author(s):  
Saskia Grootemaat ◽  
Ian J. Wright ◽  
Peter M. van Bodegom ◽  
Johannes H. C. Cornelissen ◽  
Veronica Shaw
Keyword(s):  
Forest Tree ◽  
Remarkable Feature ◽  
Interspecific Differences ◽  
Eucalypt Plantations ◽  
Physical Traits ◽  
Key Driver ◽  
Multiple Species ◽  
Species Specific ◽  
First Time ◽  
Different Tissues

Bark shedding is a remarkable feature of Australian trees, yet relatively little is known about interspecific differences in bark decomposability and flammability, or what chemical or physical traits drive variation in these properties. We measured the decomposition rate and flammability (ignitibility, sustainability and combustibility) of bark from 10 common forest tree species, and quantified correlations with potentially important traits. We compared our findings to those for leaf litter, asking whether the same traits drive flammability and decomposition in different tissues, and whether process rates are correlated across tissue types. Considerable variation in bark decomposability and flammability was found both within and across species. Bark decomposed more slowly than leaves, but in both tissues lignin concentration was a key driver. Bark took longer to ignite than leaves, and had longer mass-specific flame durations. Variation in flammability parameters was driven by different traits in the different tissues. Decomposability and flammability were each unrelated, when comparing between the different tissue types. For example, species with fast-decomposing leaves did not necessarily have fast-decomposing bark. For the first time, we show how patterns of variation in decomposability and flammability of bark diverge across multiple species. By taking species-specific bark traits into consideration there is potential to make better estimates of wildfire risks and carbon loss dynamics. This can lead to better informed management decisions for Australian forests, and eucalypt plantations, worldwide.

scholarly journals Recognition Pattern of the Fasciola hepatica Excretome/Secretome during the Course of an Experimental Infection in Sheep by 2D Immunoproteomics

Pathogens ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 725
Author(s):  
David Becerro-Recio ◽  
Javier González-Miguel ◽  
Alberto Ucero ◽  
Javier Sotillo ◽  
Álvaro Martínez-Moreno ◽  
...  
Keyword(s):  
Experimental Infection ◽  
Fasciola Hepatica ◽  
Cathepsin L ◽  
Helminth Parasites ◽  
Glutathione S Transferase ◽  
Serum Samples ◽  
Secretory Products ◽  
Immunogenic Proteins ◽  
Host Parasite ◽  
First Time

Excretory/secretory products released by helminth parasites have been widely studied for their diagnostic utility, immunomodulatory properties, as well as for their use as vaccines. Due to their location at the host/parasite interface, the characterization of parasite secretions is important to unravel the molecular interactions governing the relationships between helminth parasites and their hosts. In this study, the excretory/secretory products from adult worms of the trematode Fasciola hepatica (FhES) were employed in a combination of two-dimensional electrophoresis, immunoblot and mass spectrometry, to analyze the immune response elicited in sheep during the course of an experimental infection. Ten different immunogenic proteins from FhES recognized by serum samples from infected sheep at 4, 8, and/or 12 weeks post-infection were identified. Among these, different isoforms of cathepsin L and B, peroxiredoxin, calmodulin, or glutathione S-transferase were recognized from the beginning to the end of the experimental infection, suggesting their potential role as immunomodulatory antigens. Furthermore, four FhES proteins (C2H2-type domain-containing protein, ferritin, superoxide dismutase, and globin-3) were identified for the first time as non-immunogenic proteins. These results may help to further understand host/parasite relationships in fasciolosis, and to identify potential diagnostic molecules and drug target candidates of F. hepatica.

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