scholarly journals Analysis of Acanthamoeba genotypes from public freshwater sources in Thailand reveals a new genotype, T23 Acanthamoeba bangkokensis sp. nov.

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Chaturong Putaporntip ◽  
Napaporn Kuamsab ◽  
Warisa Nuprasert ◽  
Rattanaporn Rojrung ◽  
Urassaya Pattanawong ◽  
...  
Keyword(s):  
Phylogenetic Trees ◽  
18S Rrna ◽  
Nucleotide Diversity ◽  
Purifying Selection ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Group Iii ◽  
Subtype B ◽  
New Genotype ◽  
Novel Genotype

AbstractA survey of Acanthamoeba in 100 public freshwater sources in 28 provinces across Thailand has identified 9 genotypes comprising T2/6, T3-T5, T9, T11, T12, T18 and a novel ‘T23’ among 131 isolates. Sequencing of the near complete 18S rRNA gene of Acanthamoeba of all isolates has shown that the most predominant genotype T4 found in 87 isolates (66.4%) contained 4 subtypes, i.e. T4A, T4B, T4C and T4F, while all isolates assigned to genotype T2/6 belonged to subtype B. Among intron-bearing genotypes, most isolates harbouring genotype T3 contained S516 introns, characterised by 3 distinct variants whilst all genotypes T4A and T5 were intronless. Identical 18S rRNA sequences of Acanthamoeba were identified across regions of the country and four isolates in this study shared the same sequences with those from remote nations, suggesting that some strains have reproductive success in diverse ecological niche. Nucleotide diversity of genotypes T2/6B, T3, T4, T9 and T11 in this study was significantly less than that among global isolates outside Thailand, implying that limited sequence diversity occurred within local populations. A remarkably higher level of nucleotide diversity in genotype T11 than those of other genotypes (0.041 vs. 0.012–0.024) could be due to cryptic subtypes. Recombination breakpoints have been detected within genotypes and subtypes as well as within isolates despite no evidence for sexual and parasexual cycles in the genus Acanthamoeba. Tajima’s D, Fu & Li’s D* and F* statistics revealed significantly negative deviation from neutrality across genotypes and subtypes, implying purifying selection in this locus. The 18S rRNA gene of the novel genotype ‘T23’ displayed 7.82% to 28.44% sequence differences in comparison with all known genotypes. Both Bayesian and maximum likelihood phylogenetic trees have placed genotype T23 as sister to the clade comprising genotypes T10, T12 and T14, all of these possess cyst structure belonging to morphological group III. Hence, Acanthamoeba bangkokensis sp. nov. is proposed for this novel genotype. It is likely that more genotypes of Acanthamoeba remain to be discovered while the evolution of the 18S rRNA gene of this pathogenic-free living amoeba seems to be ongoing.

scholarly journals Analyses of separate and concatenated cox1 and 18S rRNA gene sequences indicate that the bat piroplasm Babesia vesperuginis is phylogenetically close to Cytauxzoon felis and the ‘prototheilerid’ Babesia conradae

2018 ◽  
Vol 66 (1) ◽  
pp. 107-115 ◽  
Author(s):  
Sándor Hornok ◽  
Alexandra Corduneanu ◽  
Jenő Kontschán ◽  
Katinka Bekő ◽  
Krisztina Szőke ◽  
...  
Keyword(s):  
Phylogenetic Trees ◽  
18S Rrna ◽  
Phylogenetic Analyses ◽  
18S Rrna Gene ◽  
Sensu Stricto ◽  
Rrna Gene ◽  
Sequence Comparisons ◽  
Important Species ◽  
Cytauxzoon Felis ◽  
Phylogenetic Status

Babesia vesperuginis is the only piroplasm known to infect bats. Unlike most members of the genus Babesia, it is probably transmitted by a soft tick species (i.e. Argas vespertilionis). Recently, two studies have been conducted to clarify the phylogenetic status of this species, and both agreed on placing it into a basal position among Babesia sensu stricto (s.s.). However, several important groups of piroplasms were not included in the already reported phylogenetic trees of B. vesperuginis isolates. Therefore, the aim of the present study was to amplify an approx. 950-bp fragment of the cytochrome c oxidase subunit 1 (cox1) gene of B. vesperuginis from A. vespertilionis specimens, and to compare its sequences with those from other piroplasmid groups in a broader phylogenetic context. Sequence comparisons focusing on either 18S rRNA or cox1 genes, as well as phylogenetic analyses involving separate and concatenated 18S rRNA and cox1 sequences indicate that B. vesperuginis is more closely related to the phylogenetic group of Theileriidae than to Babesia s.s. In particular, B. vesperuginis clustered closest to Cytauxzoon felis and the ‘prototheilerid’ B. conradae. The results of this study highlight that B. vesperuginis is a unique and taxonomically important species, which should be included in future studies aimed at resolving the comprehensive phylogeny of Piroplasmida.

scholarly journals Comparison of potential diatom ‘barcode’ genes (the 18S rRNA gene and ITS, COI, rbcL) and their effectiveness in discriminating and determining species taxonomy in the Bacillariophyta

2015 ◽  
Vol 65 (Pt_4) ◽  
pp. 1369-1380 ◽  
Author(s):  
Liliang Guo ◽  
Zhenghong Sui ◽  
Shu Zhang ◽  
Yuanyuan Ren ◽  
Yuan Liu
Keyword(s):  
Phylogenetic Trees ◽  
18S Rrna ◽  
Mitochondrial Gene ◽  
Large Subunit ◽  
18S Rrna Gene ◽  
Marine Ecology ◽  
The Other ◽  
Rrna Genes ◽  
Rrna Gene ◽  
Potential Marker

Diatoms form an enormous group of photoautotrophic micro-eukaryotes and play a crucial role in marine ecology. In this study, we evaluated typical genes to determine whether they were effective at different levels of diatom clustering analysis to assess the potential of these regions for barcoding taxa. Our test genes included nuclear rRNA genes (the nuclear small-subunit rRNA gene and the 5.8S rRNA gene+ITS-2), a mitochondrial gene (cytochrome c-oxidase subunit 1, COI), a chloroplast gene [ribulose-1,5-biphosphate carboxylase/oxygenase large subunit (rbcL)] and the universal plastid amplicon (UPA). Calculated genetic divergence was highest for the internal transcribed spacer (ITS; 5.8S+ITS-2) (p-distance of 1.569, 85.84 % parsimony-informative sites) and COI (6.084, 82.14 %), followed by the 18S rRNA gene (0.139, 57.69 %), rbcL (0.120, 42.01 %) and UPA (0.050, 14.97 %), which indicated that ITS and COI were highly divergent compared with the other tested genes, and that their nucleotide compositions were variable within the whole group of diatoms. Bayesian inference (BI) analysis showed that the phylogenetic trees generated from each gene clustered diatoms at different phylogenetic levels. The 18S rRNA gene was better than the other genes in clustering higher diatom taxa, and both the 18S rRNA gene and rbcL performed well in clustering some lower taxa. The COI region was able to barcode species of some genera within the Bacillariophyceae. ITS was a potential marker for DNA based-taxonomy and DNA barcoding of Thalassiosirales, while species of Cyclotella, Skeletonema and Stephanodiscus gathered in separate clades, and were paraphyletic with those of Thalassiosira. Finally, UPA was too conserved to serve as a diatom barcode.

scholarly journals Genetic diversity within the 18S rRNA and actin locus of Cryptosporidium scrofarum (Apicomplexa: Cryptosporidiidae) infecting domestic pigs (Sus scrofa domesticus) of India

2021 ◽  
Vol 91 (3) ◽  
pp. 269-276
Author(s):  
Devina Sharma ◽  
◽  
Nirbhay K. Singh ◽  
Harkirat Singh ◽  
Shitanshu S. Rath ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Sus Scrofa ◽  
18S Rrna ◽  
Pcr Amplification ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Domestic Pigs ◽  
New Genotype ◽  
Sus Scrofa Domesticus ◽  
Genotype Ii

The genetic diversity was studied of Cryptosporidium scrofarum (syn Cryptosporidium pig genotype II) of domestic pigs (Sus scrofa domesticus) from Punjab, India. Nested PCR amplification targeting the 18S rRNA and actin gene loci from Cryptosporidium positive samples was carried out, and the amplicons were sequenced. Phylogenetic comparison of a partial 18S rRNA gene revealed that they were genetically most similar to C. scrofarum isolated from other parts of the world. However, comparison of sequences representing a fragment of the genomic actin locus identified a new genotype conserved within the isolates sampled from India but distinct from other published sequences, suggesting the presence of a different Indian genotype.

scholarly journals New Hosts ofSimplicimonas similisandTrichomitus batrachorumIdentified by 18S Ribosomal RNA Gene Sequences

2013 ◽  
Vol 2013 ◽  
pp. 1-5 ◽  
Author(s):  
Kris Genelyn B. Dimasuay ◽  
Orlie John Y. Lavilla ◽  
Windell L. Rivera
Keyword(s):  
Phylogenetic Trees ◽  
18S Rrna ◽  
18S Rrna Gene ◽  
Bootstrap Support ◽  
Rrna Gene ◽  
Gene Sequences ◽  
Pathogenic Potential ◽  
Gene Sequence Analysis ◽  
High Bootstrap ◽  
Recent Classification

Trichomonads are obligate anaerobes generally found in the digestive and genitourinary tract of domestic animals. In this study, four trichomonad isolates were obtained from carabao, dog, and pig hosts using rectal swab. Genomic DNA was extracted using Chelex method and the 18S rRNA gene was successfully amplified through novel sets of primers and undergone DNA sequencing. Aligned isolate sequences together with retrieved 18S rRNA gene sequences of known trichomonads were utilized to generate phylogenetic trees using maximum likelihood and neighbor-joining analyses. Two isolates from carabao were identified asSimplicimonas similiswhile each isolate from dog and pig was identified asPentatrichomonas hominisandTrichomitus batrachorum, respectively. This is the first report ofS. similisin carabao and the identification ofT. batrachorumin pig using 18S rRNA gene sequence analysis. The generated phylogenetic tree yielded three distinct groups mostly with relatively moderate to high bootstrap support and in agreement with the most recent classification. Pathogenic potential of the trichomonads in these hosts still needs further investigation.

scholarly journals Molecular survey of coccidian infections of the side-blotched lizard Uta stansburiana on San Benito Oeste Island, Mexico

Parasite ◽  
2018 ◽  
Vol 25 ◽  
pp. 43
Author(s):  
Petra Quillfeldt ◽  
Tanja Romeike ◽  
Juan F. Masello ◽  
Gerald Reiner ◽  
Hermann Willems ◽  
...  
Keyword(s):  
Phylogenetic Trees ◽  
18S Rrna ◽  
Phylogenetic Analyses ◽  
Genetic Network ◽  
18S Rrna Gene ◽  
Blood Parasites ◽  
Rrna Gene ◽  
Uta Stansburiana ◽  
High Prevalence ◽  
High Uncertainty

Blood parasites are found in many vertebrates, but the research on blood parasites of lizards is still at its onset. We analyzed blood samples from side-blotched lizards Uta stansburiana from San Benito Oeste Island, Mexico, to test for the presence of hemoparasites. We found a high prevalence (23 out of 27 samples) of a blood parasite of the genus Lankesterella (Coccidia, Eimeriorina, Lankesterellidae) according to phylogenetic analyses of the parasite 18S rRNA gene. Similar parasites (97–99% similarity) have recently been described for Uta stansburiana from California. The parasite 18S rRNA gene showed high variability, both within San Benito and compared to California. The next closest matches of the parasite DNA with 97–98% similarity included a range of different genera (Lankesterella, Schellackia, Eimeria, Isospora and Caryospora). A high uncertainty in the deeper branches of the phylogenetic trees, and many missing links in genetic network analysis, were in line with previous suggestions that the coccidians are an understudied group with large knowledge gaps in terms of their diversity and taxonomy. Further studies are needed to resolve the evolutionary relationships within the Eimeriorina.

scholarly journals Ecological and evolutionary patterns in the enigmatic protist genus Percolomonas (Heterolobosea; Discoba) from diverse habitats

2019 ◽  
Author(s):  
Denis V. Tikhonenkov ◽  
Soo Hwan Jhin ◽  
Yana Eglit ◽  
Kai Miller ◽  
Andrey Plotnikov ◽  
...  
Keyword(s):  
18S Rdna ◽  
Phylogenetic Trees ◽  
18S Rrna ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Heterotrophic Flagellate ◽  
Evolutionary Patterns ◽  
The Usa ◽  
Saline Habitats ◽  
Morphological Differences

AbstractThe heterotrophic flagellate Percolomonas cosmopolitus (Heterolobosea) is often observed in saline habitats worldwide that range from coastal waters to saturated brines. However, only two cultures assigned to this morphospecies have been examined using molecular methods, and their 18S rRNA gene sequences are extremely different. Further the salinity tolerances of individual strains are unknown. Thus, our knowledge on the autecology and diversity in this morphospecies is deficient. Here, we report 18S rRNA gene data on seven strains similar to P. cosmopolitus from seven geographically remote locations (New Zealand, Kenya, Korea, Poland, Russia, Spain, and the USA) with sample salinities ranging from 4‰ to 280‰, and compare morphology and salinity tolerance of the nine available strains. Percolomonas cosmopolitus-like strains show few-to-no consistent morphological differences, and form six clades separated by often extremely large 18S rDNA divergences (up to 42.4%). Some strains grew best at salinities from 75 to 125‰ and represent halophiles. All but one of these belonged to two geographically heterogeneous clusters that formed a robust monophyletic group in phylogenetic trees; this likely represents an ecologically specialized subclade of halophiles. Our results suggest that P. cosmopolitus is a cluster of several cryptic species (at least), which are unlikely to be distinguished by geography. Interestingly, the 9 Percolomonas strains formed a clade in 18S rDNA phylogenies, unlike most previous analyses based on two sequences.

scholarly journals Analytical validation of a real-time hydrolysis probe PCR assay for quantifying Plasmodium falciparum parasites in experimentally infected human adults

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Claire Y. T. Wang ◽  
Emma L. Ballard ◽  
Zuleima Pava ◽  
Louise Marquart ◽  
Jane Gaydon ◽  
...  
Keyword(s):  
Clinical Trials ◽  
Plasmodium Falciparum ◽  
18S Rrna ◽  
Drug Efficacy ◽  
18S Rrna Gene ◽  
Molecular Techniques ◽  
Qpcr Assay ◽  
Rrna Gene ◽  
Analytical Sensitivity ◽  
Blood Samples

Abstract Background Volunteer infection studies have become a standard model for evaluating drug efficacy against Plasmodium infections. Molecular techniques such as qPCR are used in these studies due to their ability to provide robust and accurate estimates of parasitaemia at increased sensitivity compared to microscopy. The validity and reliability of assays need to be ensured when used to evaluate the efficacy of candidate drugs in clinical trials. Methods A previously described 18S rRNA gene qPCR assay for quantifying Plasmodium falciparum in blood samples was evaluated. Assay performance characteristics including analytical sensitivity, reportable range, precision, accuracy and specificity were assessed using experimental data and data compiled from phase 1 volunteer infection studies conducted between 2013 and 2019. Guidelines for validation of laboratory-developed molecular assays were followed. Results The reportable range was 1.50 to 6.50 log10 parasites/mL with a limit of detection of 2.045 log10 parasites/mL of whole blood based on a parasite diluted standard series over this range. The assay was highly reproducible with minimal intra-assay (SD = 0.456 quantification cycle (Cq) units [0.137 log10 parasites/mL] over 21 replicates) and inter-assay (SD = 0.604 Cq units [0.182 log10 parasites/mL] over 786 qPCR runs) variability. Through an external quality assurance program, the QIMR assay was shown to generate accurate results (quantitative bias + 0.019 log10 parasites/mL against nominal values). Specificity was 100% after assessing 164 parasite-free human blood samples. Conclusions The 18S rRNA gene qPCR assay is specific and highly reproducible and can provide reliable and accurate parasite quantification. The assay is considered fit for use in evaluating drug efficacy in malaria clinical trials.

scholarly journals 16S rRNA gene and 18S rRNA gene diversity in microbial mat communities in meltwater ponds on the McMurdo Ice Shelf, Antarctica

Polar Biology ◽  
2021 ◽  
Author(s):  
Eleanor E. Jackson ◽  
Ian Hawes ◽  
Anne D. Jungblut
Keyword(s):  
16S Rrna ◽  
18S Rrna ◽  
High Throughput Sequencing ◽  
Microbial Mats ◽  
Gene Diversity ◽  
Salinity Gradient ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Ice Shelf ◽  
The Impact

AbstractThe undulating ice of the McMurdo Ice Shelf, Southern Victoria Land, supports one of the largest networks of ice-based, multiyear meltwater pond habitats in Antarctica, where microbial mats are abundant and contribute most of the biomass and biodiversity. We used 16S rRNA and 18S rRNA gene high-throughput sequencing to compare variance of the community structure in microbial mats within and between ponds with different salinities and pH. Proteobacteria and Cyanobacteria were the most abundant phyla, and composition at OTU level was highly specific for the meltwater ponds with strong community sorting along the salinity gradient. Our study provides the first detailed evaluation of eukaryote communities for the McMurdo Ice Shelf using the 18S rRNA gene. They were dominated by Ochrophyta, Chlorophyta and Ciliophora, consistent with previous microscopic analyses, but many OTUs belonging to less well-described heterotrophic protists from Antarctic ice shelves were also identified including Amoebozoa, Rhizaria and Labyrinthulea. Comparison of 16S and 18S rRNA gene communities showed that the Eukaryotes had lower richness and greater similarity between ponds in comparison with Bacteria and Archaea communities on the McMurdo Ice shelf. While there was a weak correlation between community dissimilarity and geographic distance, the congruity of microbial assemblages within ponds, especially for Bacteria and Archaea, implies strong habitat filtering in ice shelf meltwater pond ecosystems, especially due to salinity. These findings help to understand processes that are important in sustaining biodiversity and the impact of climate change on ice-based aquatic habitats in Antarctica.

scholarly journals 18S rRNA gene sequences of leptocephalus gut contents, particulate organic matter, and biological oceanographic conditions in the western North Pacific

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Tsuyoshi Watanabe ◽  
Satoshi Nagai ◽  
Yoko Kawakami ◽  
Taiga Asakura ◽  
Jun Kikuchi ◽  
...  
Keyword(s):  
Organic Matter ◽  
Particulate Organic Matter ◽  
North Pacific ◽  
Western North Pacific ◽  
18S Rrna ◽  
18S Rrna Gene ◽  
Gut Contents ◽  
Rrna Gene ◽  
Marine Snow ◽  
Chlorophyll Maximum

AbstractEel larvae apparently feed on marine snow, but many aspects of their feeding ecology remain unknown. The eukaryotic 18S rRNA gene sequence compositions in the gut contents of four taxa of anguilliform eel larvae were compared with the sequence compositions of vertically sampled seawater particulate organic matter (POM) in the oligotrophic western North Pacific Ocean. Both gut contents and POM were mainly composed of dinoflagellates as well as other phytoplankton (cryptophytes and diatoms) and zooplankton (ciliophoran and copepod) sequences. Gut contents also contained cryptophyte and ciliophoran genera and a few other taxa. Dinoflagellates (family Gymnodiniaceae) may be an important food source and these phytoplankton were predominant in gut contents and POM as evidenced by DNA analysis and phytoplankton cell counting. The compositions of the gut contents were not specific to the species of eel larvae or the different sampling areas, and they were most similar to POM at the chlorophyll maximum in the upper part of the thermocline (mean depth: 112 m). Our results are consistent with eel larvae feeding on marine snow at a low trophic level, and feeding may frequently occur in the chlorophyll maximum in the western North Pacific.

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