Apicomplexa primers amplify Proteromonas (Stramenopiles, Slopalinida, Proteromonadidae) in tissue and blood samples from lizards

AbstractMicroscopy has traditionally been the most common method in parasitological studies, but in recent years molecular screening has become increasingly frequent to detect protozoan parasites in a wide range of vertebrate hosts and vectors. During routine molecular screening of apicomplexan parasites in reptiles using the 18S rRNA gene, we have amplified and sequenced Proteromonas parasites from three lizard hosts (less than 1% prevalence). We conducted phylogenetic analysis to confirm the taxonomic position and infer their relationships with other stramenopiles. Although our phylogeny is limited due to scarcity of molecular data on these protists, our results confirm they are closely related to Proteromonas lacertae. Our findings show that unexpected parasites can be amplified from host samples (blood and tissue) using general procedures to detect hemoparasites, and stress that positive PCR amplifications alone should not be considered as definitive proof of infection by particular parasites. Further validation by sequence confirmation and thorough phylogenetic assessment will not only avoid false positives and biased prevalence estimates but also provide valuable information on the biodiversity and phylogenetic relationships of other parasitic organisms. More generally, our results illustrate the perils of general diagnosis protocols in parasitological studies and the need of cross-validation procedures.

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

Parasitology ◽

10.1017/s0031182013001960 ◽

2013 ◽

Vol 141 (5) ◽

pp. 646-651 ◽

Cited By ~ 9

Author(s):

GASTÓN MORÉ ◽

NIKOLA PANTCHEV ◽

DALAND C. HERRMANN ◽

MAJDA GLOBOKAR VRHOVEC ◽

SABINE ÖFNER ◽

...

Keyword(s):

18S Rrna ◽

Sequence Data ◽

18S Rrna Gene ◽

Rrna Genes ◽

Rrna Gene ◽

Intermediate Hosts ◽

Apicomplexan Parasites ◽

Large Numbers ◽

Wide Range ◽

18S Rrna Genes

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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Cryptosporidium spp. in wild murids (Rodentia) from Corsica, France

Parasitology Research ◽

10.1007/s00436-021-07369-4 ◽

2021 ◽

Author(s):

Katherine García-Livia ◽

Ángela Fernández-Álvarez ◽

Carlos Feliu ◽

Jordi Miquel ◽

Yann Quilichini ◽

...

Keyword(s):

Rattus Rattus ◽

18S Rrna Gene ◽

Rrna Gene ◽

Mus Musculus Domesticus ◽

Protozoan Parasites ◽

Wild Rodents ◽

Genotype Iii ◽

Mediterranean Island ◽

Murine Rodents ◽

Vertebrate Hosts

AbstractCryptosporidium spp. are worldwide protozoan parasites that can affect to a broad range of vertebrate hosts, including rodents. In the island of Corsica (France), there are no previous data about these protozoa infecting wild rodents. To estimate the distribution and occurrence, a total of 117 wild murine rodents of the species Rattus rattus (84), Mus musculus domesticus (21), Apodemus sylvaticus (11), and Rattus norvegicus (1) were captured in 24 different biotopes. Fecal samples were screened for Cryptosporidium spp. by nested PCR to amplify an 830 bp fragment of the 18S rRNA gene. As general occurrence, 15.4% of the rodents analyzed were positive for Cryptosporidium spp., being detected widely distributed along the island in R. rattus (17.6%) and M. m. domesticus (14.3%). Cryptosporidium viatorum, Cryptosporidium sp. rat genotype II, and Cryptosporidium sp. rat genotype III were successfully identified in R. rattus. The results herein reported provide the first data on Cryptosporidium spp. in wild murine species from a Mediterranean island and constitute the first report of the zoonotic species C. viatorum in R. rattus. Although a low occurrence of Cryptosporidium spp. in murids was obtained and only in one animal the zoonotic species C. viatorum was identified, our results highlight that wild murine rodents from Corsica could mediate in the maintenance and transmission of this protozoan to the environment and other hosts including humans and animals. Further studies are required to better understand the epidemiology of Cryptosporidium spp. in wild rodents from Corsica and their possible public health repercussions.

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Molecular Detection ofTheileriaspp. in Livestock on Five Caribbean Islands

BioMed Research International ◽

10.1155/2015/624728 ◽

2015 ◽

Vol 2015 ◽

pp. 1-8 ◽

Cited By ~ 9

Author(s):

Jilei Zhang ◽

Patrick Kelly ◽

Jing Li ◽

Chuanling Xu ◽

Chengming Wang

Keyword(s):

Molecular Detection ◽

18S Rrna Gene ◽

Rrna Gene ◽

Limited Information ◽

Protozoan Parasites ◽

Sheep And Goats ◽

Caribbean Islands ◽

Related Organism ◽

Wide Range ◽

The Caribbean

Theileriaspp. are tick-transmitted, intracellular apicomplexan protozoan parasites infecting a wide range of animals. As there is very limited information on the prevalence ofTheileriaspp. in the Caribbean we used the recently described genus-specific pan-TheileriaFRET-qPCR to identify infected animals in the region and a standard 18S rRNA gene PCR and sequencing to determine the species involved. We foundTheileriaspp. in 9% of the convenience samples of animals (n=752) studied from five Caribbean islands. Donkeys (20.0%: 5/25) were most commonly infected, followed by sheep (17.4%, 25/144), cattle (6.8%; 22/325), goats (5.0%; 12/238), and horses (5.0%; 1/20). Six species ofTheileriawere identified:T.equi(donkeys, cattle, goats, and sheep),Theileriasp. OT3 (sheep and goats),Theileriasp. NG-2013a (cattle),Theileriasp. YW-2014 (donkeys),Theileriasp. B15a (goats), andBabesia vulpesor a closely related organism (sheep and goats). OnlyT. equihas been previously reported in the Caribbean. Our findings expand the known host ranges ofTheileriaspp. and the known distribution of the organisms around the world.

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Morphological redescription and molecular characterization of Trichodina matsu Basson & Van As, 1994 (Ciliophora, Mobilida, Trichodinidae) infecting Tachysurus fulvidraco (Richardson, 1846) from Chongqing, China

Zootaxa ◽

10.11646/zootaxa.4995.2.6 ◽

2021 ◽

Vol 4995 (2) ◽

pp. 334-344

Author(s):

QIAN ZHOU ◽

FAHUI TANG ◽

YUANJUN ZHAO

Keyword(s):

Phylogenetic Analyses ◽

Molecular Data ◽

18S Rrna Gene ◽

Rrna Gene ◽

Similar Species ◽

Sequence Alignments ◽

Multiple Sequence ◽

Multiple Sequence Alignments ◽

Morphological And Molecular Data ◽

Tachysurus Fulvidraco

During a survey of parasitic ciliates in Chongqing, China, Trichodina matsu Basson & Van As, 1994 was isolated from gills of Tachysurus fulvidraco. Furthermore, the 18S rRNA gene and ITS-5.8S rRNA region of T. matsu were sequenced for the first time and applied for the species identification and comparison with similar species in the present study. Based on the morphological and molecular comparisons, the results indicate that T. matsu is an ectoparasite specific for the Siluriformes catfish. Based on the analyses of genetic distance, multiple sequence alignments, and phylogenetic analyses, no obvious differentiation within populations of T. matsu was found. In addition, the ‘Trichodina hyperparasitis’ (KX904933) in GenBank is a misidentification and appears to be conspecific with T. matsu according to the comparison of morphological and molecular data.

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Emergence of Babesia Conradae Infection in Coyote-hunting Greyhounds in Oklahoma, USA

10.21203/rs.3.rs-154040/v1 ◽

2021 ◽

Author(s):

Erin M Stayton ◽

Megan Lineberry ◽

Jennifer Thomas ◽

Tina Bass ◽

Kelly Allen ◽

...

Keyword(s):

18S Rrna ◽

Clinical Signs ◽

18S Rrna Gene ◽

Post Treatment ◽

Rrna Gene ◽

Babesia Gibsoni ◽

Wide Range ◽

Pcr Products ◽

Life Threatening ◽

Tick Vectors

Abstract Background: Babesia species are intraerythrocytic Apicomplexan parasites that infect a wide range of vertebrate hosts. These pathogens are typically transmitted either by tick vectors or by direct blood-to-blood contact, and may cause life-threatening clinical disease such as thrombocytopenia, hemolytic anemia, and acute renal failure in canine hosts. While Babesia vogeli and Babesia gibsoni infections have both been reported in Oklahoma, reports of B. conradae infections have been limited to California. Methods: Whole blood samples were collected in EDTA tubes from all dogs in four separate kennels in Oklahoma. DNA was extracted from each blood sample and a nested PCR was performed using general Apicomplexan primers for the partial 18S rRNA gene. PCR products were electrophoresed in agarose matrix and appropriately sized amplicons were sequenced. Sequences were compared to reference 18S rRNA sequences available in GenBank, and samples with >98% homology to B. conradae (GenBank MK256976) were considered positive. B. conradae positive dogs were then treated with atovaquone (13.5 mg/kg TID) and azithromycin (10 mg/kg SID) for 10 days and retested at 30 and 60 days post treatment by PCR. Results: Fifteen of 40 dogs tested positive for B. conradae with 98–100% sequence homology to B. conradae from California. All positive cases were coyote-hunting Greyhounds. Treatment of clinically ill dogs with atovaquone and azithromycin resulted in complete clinical recovery in clinically ill dogs and all treated dogs had negative follow-up PCR at 30 and 60 days post treatment. Conclusions: Collectively, this study (i) documents the occurrence of B. conradae in Oklahoma, (ii) highlights this pathogen as a differential to be considered when clinical signs are present, and (iii) supports the use of atovaquone and azithromycin as effective treatment in these cases.

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Molecular data provide new insights into the phylogeny of Cladonotinae (Orthoptera: Tetrigoidea) from China with the description of a new genus and species

Zootaxa ◽

10.11646/zootaxa.4809.3.8 ◽

2020 ◽

Vol 4809 (3) ◽

pp. 547-559

Author(s):

RONG-JIAO ZHANG ◽

CONG-LIN ZHAO ◽

FEI-PENG WU ◽

WEI-AN DENG

Keyword(s):

Ribosomal Rna ◽

Phylogenetic Relationships ◽

New Genus ◽

Molecular Data ◽

18S Rrna Gene ◽

Rrna Gene ◽

Mitochondrial Cytochrome Oxidase Subunit ◽

Close Relationship ◽

Phylogenetic Framework ◽

Mitochondrial Cytochrome Oxidase

Considerable effort has been devoted to elucidating the phylogenetic relationships of tetrigides. However, there is still no commonly accepted phylogenetic hypothesis. Therefore, the phylogenetic relationships among some subfamilies remain unclear; e.g., Cladonotinae is a controversial group, in which the phylogenetic relationships between genera and the boundaries of some of the included genera are unclear, causing some of the taxa to be difficult to identify. Therefore, an in-depth phylogenetic analysis of Cladonotinae is urgently needed. In this study, a robust phylogenetic framework for the tetrigides was reconstructed based on the combined mitochondrial cytochrome oxidase subunit I (COI), 16S ribosomal RNA (16S rRNA), and nuclear 18S ribosomal RNA (18S rRNA) gene sequences of 25 species belonging to 16 genera of Tetrigoidea from China, which included 13 species from 8 genera of Cladonotinae. Phylogenetic inferences were performed using the combined dataset and Bayesian inference (BI) and Maximum Parsimony (MP) methods, and the phylogenetic tree of Cladonotinae was reconstructed. All inferences based on the results of the present study supported the Cladonotinae subfamily as a polyphyletic group; within the Cladonotinae subfamily, Tetradinodula, and Tuberfemurus were closely related to Tetriginae, while Austrohancockia and Gibbotettix showed a close relationship to the Scelimenidae subfamily. Additionally, a new genus and new species of the Cladonotinae subfamily are described and illustrated: Hainantettix Deng, gen. nov. and Hainantettix strictivertex Deng, sp. nov.

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Review of the genus Phormidesmis (Cyanobacteria) based on environmental, morphological, and molecular data with description of a new genus Leptodesmis

Phytotaxa ◽

10.11646/phytotaxa.395.1.1 ◽

2019 ◽

Vol 395 (1) ◽

pp. 1-16 ◽

Cited By ~ 6

Author(s):

LENKA RAABOVÁ ◽

LUBOMIR KOVACIK ◽

JOSEF ELSTER ◽

OTAKAR STRUNECKÝ

Keyword(s):

Molecular Data ◽

Morphological Properties ◽

Phylogenetic Position ◽

Rrna Gene ◽

Filamentous Cyanobacteria ◽

Wide Range ◽

Significant Divergence ◽

Taxonomic Methods ◽

Morphological And Molecular Data ◽

Definition Of

Very thin filamentous cyanobacteria are ubiquitous in a wide range of environments. For many years they were traditionally studied according to their morphological properties only. With the introduction of additional taxonomic methods (cytomorphological analyses, molecular sequencing, exact ecological studies, better data about phytogeographical distribution), traditional genera such as Leptolyngbya and Phormidium were found to be polyphyletic. Phormidesmis belonged to a newly formed genus that was supposed to explain the variability of such very thin simple filamentous cyanobacteria. However, even after definition of Phormidesmis based on distinct cytomorphological and phylogenetic traits, the variability within this genus remained unresolved. Here we analyzed 26 Phormidesmis strains to describe the variability within this genus, classified two new species (P. arctica and P. communis) and transferred Leptolyngbya nigrescens into P. nigrescens. A tabular review of Phormidesmis species is included. The diacritical features of all these species are: width up to 1–4 µm, barrel-shaped cells, which can be shorter or slightly longer than wide, with apparent constrictions at the cross-walls. Our study shows that Phormidesmis is a morphologically and genetically well-defined genus with a global distribution. A newly described genus Leptodesmis has significant morphological similarities both with Phormidesmis and Leptolyngbya, however with intermediate phylogenetic position with significant divergence in 16S rRNA gene. Leptodesmis is cryptic both to Phormidesmis and Leptolyngbya. In the initial part of the life cycle resembles Leptolyngbya, the appearance of older trichomes change to Phormidesmis like morphology.

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High-resolution molecular identification of smalltooth sawfish prey

Scientific Reports ◽

10.1038/s41598-019-53931-7 ◽

2019 ◽

Vol 9 (1) ◽

Author(s):

Taylor L. Hancock ◽

Gregg R. Poulakis ◽

Rachel M. Scharer ◽

S. Gregory Tolley ◽

Hidetoshi Urakawa

Keyword(s):

18S Rrna ◽

Stomach Content Analysis ◽

18S Rrna Gene ◽

Taxonomic Resolution ◽

Molecular Technique ◽

Rrna Gene ◽

Fecal Samples ◽

Predator Prey ◽

Critically Endangered Species ◽

Wide Range

AbstractThe foundation of food web analysis is a solid understanding of predator-prey associations. Traditional dietary studies of fishes have been by stomach content analysis. However, these methods are not applicable to Critically Endangered species such as the smalltooth sawfish (Pristis pectinata). Previous research using the combination of stable isotope signatures from fin clips and 18S rRNA gene sequencing of fecal samples identified the smalltooth sawfish as piscivorous at low taxonomic resolution. Here, we present a high taxonomic resolution molecular technique for identification of prey using opportunistically acquired fecal samples. To assess potential biases, primer sets of two mitochondrial genes, 12S and 16S rRNA, were used alongside 18S rRNA, which targets a wider spectrum of taxa. In total, 19 fish taxa from 7 orders and 11 families native to the Gulf of Mexico were successfully identified. The sawfish prey comprised diverse taxa, indicating that this species is a generalist piscivore. These findings and the molecular approach used will aid recovery planning for the smalltooth sawfish and have the potential to reveal previously unknown predator-prey associations from a wide range of taxa, especially rare and hard to sample species.

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Phylogenetic information of freshwater copepod (Diaptomus sicilis) with special reference to 18S rRNA

International Journal of Biological Research ◽

10.14419/ijbr.v4i1.5836 ◽

2016 ◽

Vol 4 (1) ◽

pp. 25 ◽

Cited By ~ 1

Author(s):

Gomathi Jeyam Mookkaiah ◽

Ramanibai Ravichandran

Keyword(s):

Tamil Nadu ◽

18S Rrna ◽

Molecular Data ◽

Small Subunit ◽

18S Rrna Gene ◽

Rrna Gene ◽

Calanoid Copepods ◽

Universal Primer ◽

Pcr Products ◽

The Individual

In the present investigation to isolate freshwater calanoid copepods (Diaptomus sicilis) was characterized and identify the organisms by 18S rRNA sequencing. Plankton samples containing D. sicilis were collected during January 2014 (Post-monsoon) from Madippakkam Lake (12°57'41"N80°11'27"E) Chennai, Tamil Nadu. Immediately after sampling, specimens for genetic analyses were fixed in 95% ethyl alcohol. The total DNA was extracted from the individual copepod D. sicilis using Qiagen Blood tissue kit. The nuclear small subunit 18S rRNA gene was amplified using the Universal primer LCO —1490 (5’-GGTCAACAAATCATAAAGATATTGG-3’) and HCO-2198 (5’-TAAACTTCAGGGTGACCAAAAAATCA-3’). PCR products were loaded onto a 1% TAE agarose gel. Sequences were carried out an automated sequencer. The nucleotide sequence of 1282 base pair region of 18S rRNA was determined for D. sicilis. The similarity of sequences of D. sicilis was retrieved by BLASTn program and maximum identity and E-value was 76% and 0.00, respectively. The PCR products of D. sicilis individuals showed 80% similarity with the partial nuclear small subunit 18S rRNA gene region of other calanoid copepods. Based on molecular data the freshwater Calanoid copepods showed different algorithms and similar types of topologies useful for designing molecular analyses using phylogeny tree construction.Present molecular studies on the relationship of D. sicilis with other freshwater calanoid copepods indicate that this species is close to D. castor followed by D. keniraensis.

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