Molecular identification ofSarcocystisspp. helped to define the origin of green pythons (Morelia viridis) confiscated in Germany

SUMMARYSarcocystisspp. represent apicomplexan parasites. They usually have a heteroxenous life cycle. Around 200 species have been described, affecting a wide range of animals worldwide, including reptiles. In recent years, large numbers of reptiles have been imported into Europe as pets and, as a consequence, animal welfare and species protection issues emerged. A sample of pooled feces from four confiscated green pythons (Morelia viridis) containingSarcocystisspp. sporocysts was investigated. These snakes were imported for the pet trade and declared as being captive-bred. Full length 18S rRNA genes were amplified, cloned into plasmids and sequenced. Two differentSarcocystisspp. sequences were identified and registered asSarcocystissp. fromM. viridisin GenBank. Both showed a 95–97% sequence identity with the 18S rRNA gene ofSarcocystis singaporensis.Phylogenetic analysis positioned these sequences together with otherSarcocystisspp. from snakes and rodents as definitive and intermediate hosts (IH), respectively. Sequence data and also the results of clinical and parasitological examinations suggest that the snakes were definitive hosts forSarcocystisspp. that circulate in wild IH. Thus, it seems unlikely that the infected snakes had been legally bred. Our research shows that information on the infection of snakes withSarcocystisspp. may be used to assess compliance with regulations on the trade with wildlife species.

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Design and Validation of Four New Primers for Next-Generation Sequencing To Target the 18S rRNA Genes of Gastrointestinal Ciliate Protozoa

Applied and Environmental Microbiology ◽

10.1128/aem.01644-14 ◽

2014 ◽

Vol 80 (17) ◽

pp. 5515-5521 ◽

Cited By ~ 22

Author(s):

Suzanne L. Ishaq ◽

André-Denis G. Wright

Keyword(s):

18S Rrna ◽

18S Rrna Gene ◽

Rrna Genes ◽

Rrna Gene ◽

Content Type ◽

Hypervariable Regions ◽

Blast Database ◽

Primer Sets ◽

18S Rrna Genes ◽

Ciliate Protozoa

ABSTRACTFour new primers and one published primer were used to PCR amplify hypervariable regions within the protozoal 18S rRNA gene to determine which primer pair provided the best identification and statistical analysis. PCR amplicons of 394 to 498 bases were generated from three primer sets, sequenced using Roche 454 pyrosequencing with Titanium, and analyzed using the BLAST database (NCBI) and MOTHUR version 1.29. The protozoal diversity of rumen contents from moose in Alaska was assessed. In the present study, primer set 1, P-SSU-316F and GIC758R (amplicon of 482 bases), gave the best representation of diversity using BLAST classification, and the set amplifiedEntodinium simplexandOstracodiniumspp., which were not amplified by the other two primer sets. Primer set 2, GIC1080F and GIC1578R (amplicon of 498 bases), had similar BLAST results and a slightly higher percentage of sequences that were identified with a higher sequence identity. Primer sets 1 and 2 are recommended for use in ruminants. However, primer set 1 may be inadequate to determine protozoal diversity in nonruminants. The amplicons created by primer set 1 were indistinguishable for certain species within the generaBandia,Blepharocorys,Polycosta, andTetratoxumand betweenHemiprorodon gymnoprosthiumandProrodonopsiscoli, none of which are normally found in the rumen.

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Dogs are reservoir hosts for possible transmission of human strongyloidiasis in Thailand: molecular identification and genetic diversity of causative parasite species

Journal of Helminthology ◽

10.1017/s0022149x1900107x ◽

2019 ◽

Vol 94 ◽

Cited By ~ 3

Author(s):

O. Sanpool ◽

P.M. Intapan ◽

R. Rodpai ◽

P. Laoraksawong ◽

L. Sadaow ◽

...

Keyword(s):

18S Rrna ◽

Gastrointestinal Disease ◽

Close Association ◽

Lao Pdr ◽

Strongyloides Stercoralis ◽

18S Rrna Gene ◽

Rrna Genes ◽

Rrna Gene ◽

North East ◽

18S Rrna Genes

Abstract Human strongyloidiasis is a deleterious gastrointestinal disease mainly caused by Strongyloides stercoralis infection. We aimed to study the possible transmission of S. stercoralis between humans and pet animals. We isolated Strongyloides from humans and domestic dogs in the same rural community in north-east Thailand and compared the nucleotide sequences of derived worms using portions of the mitochondrial cytochrome c oxidase subunit 1 (cox1) and 18S ribosomal RNA (18S rRNA) genes. Twenty-eight sequences from the 18S rRNA gene were obtained from worms derived from humans (n = 23) and dogs (n = 5), and were identical with S. stercoralis sequences (from Thailand, Cambodia, Lao PDR and Myanmar) published in the GenBank database. The 28 cox1 sequences from humans and dogs showed high similarity to each other. The available published cox1 sequences (n = 150), in combination with our 28 sequences, represented 68 haplotypes distributed among four clusters. The 28 samples from the present study represented eight haplotypes including four new haplotypes. Dogs and humans shared the same haplotypes, suggesting the possibility of zoonotic transmission from pet dogs to humans. This is of concern since dogs and humans live in close association with each other.

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18S rRNA Gene Variation among Common Airborne Fungi, and Development of Specific Oligonucleotide Probes for the Detection of Fungal Isolates

Applied and Environmental Microbiology ◽

10.1128/aem.69.9.5389-5397.2003 ◽

2003 ◽

Vol 69 (9) ◽

pp. 5389-5397 ◽

Cited By ~ 62

Author(s):

Zhihong Wu ◽

Yoshihiko Tsumura ◽

Göran Blomquist ◽

Xiao-Ru Wang

Keyword(s):

18S Rrna ◽

Airborne Fungi ◽

18S Rrna Gene ◽

Rrna Genes ◽

Molecular Diagnostic ◽

Rrna Gene ◽

Oligonucleotide Probes ◽

Gene Variation ◽

Species Specific ◽

18S Rrna Genes

ABSTRACT In this study, we sequenced 18S rRNA genes (rDNA) from 49 fungal strains representing 31 species from 15 genera. Most of these species are common airborne fungi and pathogens that may cause various public health concerns. Sequence analysis revealed distinct divergence between Zygomycota and Ascomycota. Within Ascomycota, several strongly supported clades were identified that facilitate the taxonomic placement of several little-studied fungi. Wallemia appeared as the group most diverged from all the other Ascomycota species. Based on the 18S rDNA sequence variation, 108 oligonucleotide probes were designed for each genus and species included in this study. After homology searches and DNA hybridization evaluations, 33 probes were verified as genus or species specific. The optimal hybridization temperatures to achieve the best specificity for these 33 probes were determined. These new probes can contribute to the molecular diagnostic research for environmental monitoring.

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Molecular identification and phylogenetic characterisation of Thelohanellus kitauei — short communication

Acta Veterinaria Hungarica ◽

10.1556/avet.2012.055 ◽

2013 ◽

Vol 61 (1) ◽

pp. 30-35 ◽

Cited By ~ 5

Author(s):

Sang Shin ◽

Ji Kim ◽

Casiano Choresca ◽

Jee Han ◽

Jin Jun ◽

...

Keyword(s):

18S Rrna ◽

Short Communication ◽

Genetic Relationships ◽

18S Rrna Gene ◽

Rrna Genes ◽

Rrna Gene ◽

Pcr Product ◽

Cyprinus Carpio Haematopterus ◽

18S Rrna Genes ◽

Rrna Sequences

Thelohanellus kitauei was isolated from the koi Cyprinus carpio haematopterus, and the 18S rRNA gene of T. kitauei was amplified by optimised nested-PCR. The PCR product was sequenced and compared with other 18S rRNA genes of Thelohanellus species to investigate the relationships between their host specificities and infection sites. Based on the 18S rRNA sequences, T. kitauei is most closely related to T. hovorkai (which can infect the intestine). Phylogenetic analysis revealed that T. kitauei was clustered with other Thelohanellus spp. infecting Cyprininae. The present study suggests that the infection site and the host specificity (subfamily level) are reflected in the genetic relationships among Thelohanellus species.

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Phylogenetic relationships of Scelimeninae genera (Orthoptera: Tetrigoidea) based on COI, 16S rRNA and 18S rRNA gene sequences

Zootaxa ◽

10.11646/zootaxa.4482.2.11 ◽

2018 ◽

Vol 4482 (2) ◽

pp. 392

Author(s):

YA-ZHEN CHEN ◽

WEI-AN DENG ◽

JIA-MIN WANG ◽

LI-LIANG LIN ◽

SHAN-YI ZHOU

Keyword(s):

16S Rrna ◽

Genetic Distance ◽

Phylogenetic Relationships ◽

18S Rrna ◽

18S Rrna Gene ◽

Rrna Genes ◽

Rrna Gene ◽

Average Genetic Distance ◽

Molecular Phylogenetic ◽

18S Rrna Genes

Scelimeninae is an important subfamily of Tetrigoidea; however, the phylogenetic relationships within Scelimeninae are poorly understood, and its generic classification has remained unstable. In this study, the COI, 16S rRNA and 18S rRNA genes from 24 species in 9 genera within Scelimeninae were amplified and sequenced, the base composition and inter-species genetic distance of the combined sequence of COI, 16S rRNA and 18S rRNA genes were analyzed, and the molecular phylogenetic relationships were reconstructed using Maximum Likelihood (ML) and Bayesian inference (BI) methods. The results of sequence analysis showed that the total length of the combined COI, 16S rRNA and 18S rRNA gene sequence was 3507 bp, including 2345 conservative sites, 1144 variable sites and 901 parsimony-informative sites. The average A+T content was 63.5% and 78.1% in the COI, 16S rRNA sequences, respectively, indicating A+T bias. The average genetic distance between all species was 0.134, and the average genetic distance in the inner group (Scelimeninae) was 0.126. A phylogenetic tree based on the combined sequences of the COI, 16S rRNA and 18S rRNA genes showed that the phylogenetic relationships among 9 Scelimeninae genera were as follows: Criotettix + (((Zhengitettix + Hebarditettix) + (Falconius + (Scelimena + Paragavialidium))) + ((Eucriotettix + Thoradonta) + Loxilobus)). The molecular phylogenetic results generally support the morphological taxonomy; at the genus level, Criotettix, Scelimena, Paragavialidium, Thoradonta and Eucriotettix are monophyletic groups, Scelimena and Paragavialidium form sister groups, and Thoradonta and Eucriotettix also form sister groups, but the relationship between Hebarditettix and Zhengitettix needs further study. At the species level, synonyms may exist between Thoradonta spiculoba and Thoradonta transpicula and Thoradonta nodulosa and Thoradonta obtusilobata, but more studies are required to confirm this inference.

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Detection Moleculer Of Putative 18S rRNA Gen Protozoa Trichodina sp. Infected Larvae Gurami (Osphronemus gouramy L) in Balai Benih Ikan Kutasari Purbalingga Central Java

BioEksakta : Jurnal Ilmiah Biologi Unsoed ◽

10.20884/1.bioe.2020.2.3.3924 ◽

2021 ◽

Vol 3 (1) ◽

pp. 26

Author(s):

Rokhmani Rokhmani ◽

Daniel Joko Wahyono ◽

Lilis Mulyani

Keyword(s):

18S Rrna ◽

18S Rrna Gene ◽

Fish Growth ◽

Rrna Genes ◽

Rrna Gene ◽

Gene Detection ◽

Central Java ◽

Polymerase Chain ◽

18S Rrna Genes

Trichodina spp. are ectoparasitic pathogens of ciliata group that commonly infect both freshwater and marine fish, including gouramy fish. As a result of infection of Trichodina spp. this will lead to inhibition of fish growth and decreased fish production, resulting in low fish selling value. The rate of occurrence of Trichodina spp. that infects gurami can reach 100%. Research has been conducted to determine which one Trichodina spp. Protozoa that infects the gouramy seeds of BBI (Fish Seed Center) Kutasari Purbalingga following detection of 18S RNA gene. Gene detection method used in this research is Polymerase Chain Reaction (PCR) is a technique of DNA synthesis and amplification in vitro. This research is done following these methodes: (1) sampling of Gurami fish with purposive sampling which obtained from BBI Kutasari Purbalingga, (2) isolation of Trichodina spp., (3). Preparation of Trichodina spp. sample and its identification, and (4). Molecular character obervation following detection of 18S rRNA gene. This study obtained 10% percentage of detection of 18S rRNA genes of the species of Trichodina paraheterodentata that infect on the gouramy fish of Purbalingga. The percentage rate of detection of these genes is low when compared with the results of the detection of 18S rRNA Trichodina paraheterodentata gene that infects gouramy fish in Banjarnegara.

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A morphological and molecular reinvestigation of Janickina pigmentifera (Grassi, 1881) Chatton 1953 – an amoebozoan parasite of arrow-worms (Chaetognatha)

INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY ◽

10.1099/ijsem.0.005094 ◽

2021 ◽

Vol 71 (11) ◽

Author(s):

Ekaterina Volkova ◽

Alexander Kudryavtsev

Keyword(s):

18S Rrna ◽

Phylogenetic Analyses ◽

18S Rrna Gene ◽

Rrna Genes ◽

Rrna Gene ◽

Intracellular Symbiont ◽

Microscopic Features ◽

Parasitic Lifestyle ◽

18S Rrna Genes ◽

Arrow Worms

Amoebozoan parasites of arrow-worms (Chaetognatha) were isolated from their hosts living in plankton of the Bay of Villefranche (Mediterranean Sea). Based on the light microscopic characters, the amoebae were identified as Janickina pigmentifera (Grassi, 1881) by their limax locomotive form and due to the presence of the intracellular symbiont, Perkinsela amoebae, surrounded by a layer of pigment granules. Sequences of the 18S rRNA gene of both J. pigmentifera and its symbiont were obtained for the first time. The molecular phylogenetic analyses of 18S rRNA gene placed J. pigmentifera within the genus Neoparamoeba, a taxon also characterized by the presence of a symbiont, known as Perkinsela amoebae-like organism (PLO). The 18S rRNA gene sequence of P. amoebae from J. pigmentifera grouped with the sequences of 18S rRNA genes of PLOs from Neoparamoeba branchiphila and Neoparamoeba invadens. The first photo documentation of the light microscopic features of J. pigmentifera, such as locomotive form, the morphology of the nucleus and P. amoebae have been provided. The new results support the affinity of J. pigmentifera with the family Paramoebidae suggested previously based on the presence of PLO. In contrast to Janickina, typical members of Paramoebidae (Neoparamoeba and Paramoeba) have a flattened, dactylopodial locomotive form. This discrepancy in morphology can be explained by the obligate parasitic lifestyle of Janickina.

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Theileria ovis (Piroplasmida: Theileriidae) Detected in Melophagus ovinus (Diptera: Hippoboscoidea) and Ornithodoros lahorensis (Ixodida: Argasidae) Removed From Sheep in Xinjiang, China

Journal of Medical Entomology ◽

10.1093/jme/tjz193 ◽

2019 ◽

Vol 57 (2) ◽

pp. 631-635 ◽

Cited By ~ 1

Author(s):

Li Zhao ◽

Jinling Wang ◽

Yulin Ding ◽

Kairui Li ◽

Bo He ◽

...

Keyword(s):

18S Rrna ◽

18S Rrna Gene ◽

Economic Losses ◽

Rrna Gene ◽

Protozoan Parasites ◽

Autonomous Region ◽

Apicomplexan Parasites ◽

Wide Range ◽

Melophagus Ovinus ◽

Theileria Spp

Abstract Theileria spp. are tick-transmitted, intracellular apicomplexan protozoan parasites that infect a wide range of animals and, as such, can cause significant economic losses. The aim of the present study was to detect and analyze apicomplexan parasites from two different ectoparasites that were collected from Xinjiang Uygur Autonomous Region, China. The PCR-based detection of 18S rRNA indicated that Ornithodoros lahorensis specimens from Kashgar, Xinjiang, and Aksu were positive for Theileria spp., as were Melophagus ovinus specimens from Aksu. Meanwhile, phylogenetic analysis, based on the 18S rRNA gene sequences, revealed that the four amplified Theileria sequences could be attributed to T. ovis. To the best of our knowledge, this is the first study to report the detection of T. ovis DNA in M. ovinus and the first molecular identification study to confirm the detection of T. ovis in O. lahorensis in China. Accordingly, the present study extends the known distribution of T. ovis.

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Molecular Characteristics of Indonesian Isolate Enterocytozoon hepatopenaei Based on Sequence Analysis of 18S rRNA Genes

Omni-Akuatika ◽

10.20884/1.oa.2019.15.1.694 ◽

2019 ◽

Vol 15 (1) ◽

pp. 93

Author(s):

Yohanes Kristiawan Artanto ◽

Slamet Budi Prayitno ◽

Sarjito Sarjito ◽

Desrina Desrina ◽

Alfabetian Harjuna Condro Haditomo

Keyword(s):

Sequence Homology ◽

18S Rrna ◽

18S Rrna Gene ◽

Rrna Genes ◽

Molecular Characteristics ◽

Rrna Gene ◽

Multiple Sequence ◽

Rrna Gene Sequencing ◽

Enterocytozoon Hepatopenaei ◽

18S Rrna Genes

ABSTRACT Enterocytozoon hepatopenaei (EHP) infection have been reported as an obstacle of whiteleg shrimp (Penaeus vannamei) culture in Indonesia. However, understanding of the molecular characteristics of EHP species in Indonesia is not widely known. The aims of this study were to determine the identity and characters of DNA, and their phylogeny of EHP species from several different locations in Indonesia with specific references to 18S rRNA gene. The EHPs were collected from cultured P.vannamei from Lampung, Pangandaran, Sidoarjo, Banyuwangi, Probolinggo, Blitar, Makassar, and Lombok. Thirteen (13) samples were randomly selected to explore their gene characters through 18S rRNA gene sequencing. The primers used were EHP_F and EHP_1R. Parameter observed were DNA sequencing, nucleotide sequence homology with related available genes in the Gen Bank database, multiple sequence alignment, and reconstruction of genetic relationship trees. DNA sequence homology analysis showed that all samples had 99.89-100% similarity to Indian Enterocytozoon hepatopenaei (Accession Number MH259890.1 and MH260592.1). The alignment results illustrated that all EHP sequences of Indonesian isolates were 100% identical each other. The phylogenetic tree topology provided information that all sample accessions were in the same clade and spread evenly. The conclusion were that the Indonesian EHP species were identical (100%) and it could be said that they were genetically homogeneous. Keywords: Enterocytozoon hepatopenaei, 18S rRNA gene, Indonesia.

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Small subunit (18S) ribosomal RNA gene divergence in the genus Schistosoma

Parasitology ◽

10.1017/s0031182000067251 ◽

1993 ◽

Vol 107 (2) ◽

pp. 147-156 ◽

Cited By ~ 32

Author(s):

D. A. Johnston ◽

R. A. Kane ◽

D. Rollinson

Keyword(s):

Ribosomal Rna ◽

18S Rrna ◽

Gene Sequence ◽

Small Subunit ◽

18S Rrna Gene ◽

Rrna Genes ◽

Rrna Gene ◽

18S Ribosomal Rna Gene ◽

Species Groups ◽

18S Rrna Genes

SUMMARYAn entire 18S rRNA gene sequence from Schistosoma spindale (1990 bases) and partial 18S rRNA gene sequences from S. haematobium (1950 bases) and S. japonicum (1648 bases) have been determined. Together with the previously published sequence for the S. mansoni 18S rRNA gene, these data encompass the 4 recognized Schistosoma species groups. Although Schistosoma 18S rRNA genes are highly conserved, the sequences permit a preliminary molecular phylogeny to be established for the genus. This identifies S. haematobium and S. spindale as sister taxa in a clade with S. mansoni. S. japonicum does not appear to be closely related to this clade. Much of the observed variation occurs within a ‘hypervariable’ stretch of the gene corresponding to part of the V4 region of 18S rRNA. Despite this variation, the 3 new sequences fit models of 18S rRNA secondary structure predicted from the S. mansoni sequence.The sequences reported in this paper have been submitted to the EMBL Database under accession numbers Z11976 (S. haematobium), Z11590 (S. japonicum) and Z11979 (S. spindale).

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