Prototheca bovis, a unicellular achlorophyllous trebouxiophyte green alga in the healthy human intestine

Introduction. Prototheca species are non-photosynthetic trebouxiophyte algae ubiquitously distributed in nature and can be found in sewage and soil. This microbial eukaryote causes human protothecosis in immunocompromised individuals. Thus, Prototheca presence in the stool of individuals without gastrointestinal symptoms has been reported only rarely. Hypothesis/Gap statement. There is an absence of detailed characterization of human Prototheca isolates. Aim. The aim of this study was to perform morphological and molecular characterization of Prototheca isolates obtained from human stool. Methodology. Prototheca was isolated from faecal samples of four individuals living in a rural area in Thailand. A combination of bioimaging along with molecular and bioinformatics tools was used to characterize the four strains. The growth rate was tested using four media and three temperature conditions. Phylogenetic analysis using the small subunit ribosomal RNA (SSU rRNA) and cytochrome b (cytb) was also performed. Results. Static and live microscopy demonstrated the various life stages of Prototheca and its major defining cellular characteristics. An optimized DNA extraction methodology that improves DNA yield is provided. Partial fragments of the SSU rRNA and cytb genes were obtained. Phylogenetic analysis placed all four strains in the clade with Prototheca bovis. More broadly, Prototheca was not monophyletic but split into at least two distinct clades instead. Conclusion. The results represent the first molecular characterization of Prototheca in Thailand. The study provides insight into transmission dynamics of the organism and potential caveats in estimating the global prevalence of Prototheca. These will spearhead further investigations on Prototheca occurrence in rural areas of both industrialized and developing nations.

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Molecular Characterization of Cryptosporidium molnari Reveals a Distinct Piscine Clade

Applied and Environmental Microbiology ◽

10.1128/aem.01335-10 ◽

2010 ◽

Vol 76 (22) ◽

pp. 7646-7649 ◽

Cited By ~ 25

Author(s):

Oswaldo Palenzuela ◽

Pilar Alvarez-Pellitero ◽

Ariadna Sitjà-Bobadilla

Keyword(s):

Phylogenetic Analysis ◽

Molecular Characterization ◽

Distinct Species ◽

Small Subunit ◽

Ssu Rrna

ABSTRACT Multilocus phylogenetic analysis of small-subunit (SSU) rRNA and actin from Cryptosporidium molnari clustered this species with the C. molnari-like genotype of an isolate from the guppy, although the two fish isolates seem to be distinct species. The analysis of available piscine genotypes provides some support for cladistic congruence of the genus Piscicryptosporidium, but additional piscine genotypes are needed.

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Prevalence and molecular characterization of Cryptosporidium spp. in Père David’s deer (Elaphurus davidianus) in Jiangsu, China

Revista Brasileira de Parasitologia Veterinária ◽

10.1590/s1984-29612020013 ◽

2020 ◽

Vol 29 (2) ◽

Author(s):

Si-Yang Huang ◽

Yi-Min Fan ◽

Yi Yang ◽

Yi-Jun Ren ◽

Jing-Zhi Gong ◽

...

Keyword(s):

Molecular Characterization ◽

Small Subunit ◽

Basic Knowledge ◽

Rrna Gene ◽

Ssu Rrna ◽

Ssu Rrna Gene ◽

Cryptosporidium Spp ◽

Père David’S Deer ◽

Rrna Gene Sequencing

Abstract Cryptosporidium is a zoonotic parasite that causes diarrhea in a broad range of animals, including deer. Little is known about the prevalence and genotype of Cryptosporidium spp. in Père David’s deer. In this study, 137 fecal samples from Père David’s deer were collected between July 2017 and August 2018 in the Dafeng Reserve and analyzed for Cryptosporidium spp. by nested-PCR based on the small subunit ribosomal RNA (SSU rRNA) gene, followed by sequence analyses to determine the species. The 60 kDa glycoprotein (gp60) gene was used to characterize Cryptosporidium spp. Among 137 samples, 2 (1.46%) were positive for Cryptosporidium spp. according to SSU rRNA gene sequencing results. Both samples belonged to the Cryptosporidium deer genotype, with two nucleotide deletions and one nucleotide substitution. The prevalence data and molecular characterization of this study provide basic knowledge for controlling and preventing Cryptosporidium infections in Père David’s deer in this area.

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Morphological, histological and molecular characterization of Myxidium cf. rhodei infecting the kidney of Rutilus rutilus

Diseases of Aquatic Organisms ◽

10.3354/dao03514 ◽

2020 ◽

Vol 141 ◽

pp. 39-46

Author(s):

MD Dorjievna Batueva ◽

X Pan ◽

J Zhang ◽

X Liu ◽

W Wei ◽

...

Keyword(s):

Phylogenetic Analysis ◽

Molecular Characterization ◽

Rutilus Rutilus ◽

Longitudinal Axis ◽

Suture Line ◽

Present Species ◽

Supplementary Data

In the present study, we provide supplementary data for Myxidium cf. rhodei Léger, 1905 based on morphological, histological and molecular characterization. M. cf. rhodei was observed in the kidneys of 918 out of 942 (97%) roach Rutilus rutilus (Linnaeus, 1758). Myxospores of M. cf. rhodei were fusiform with pointed ends, measuring 12.7 ± 0.1 SD (11.8-13.4) µm in length and 4.6 ± 0.1 (3.8-5.4) µm in width. Two similar pear-shaped polar capsules were positioned at either ends of the longitudinal axis of the myxospore: each of these capsules measured 4.0 ± 0.1 (3.1-4.7) µm in length and 2.8 ± 0.1 (2.0-4.0) µm in width. Polar filaments were coiled into 4 to 5 turns. Approximately 18-20 longitudinal straight ridges were observed on the myxospore surface. The suture line was straight and distinctive, running near the middle of the valves. Histologically, the plasmodia of the present species were found in the Bowman’s capsules, and rarely in the interstitium of the host. Phylogenetic analysis revealed that M. cf. rhodei was sister to M. anatidum in the Myxidium clade including most Myxidium species from freshwater hosts.

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Prevalence and molecular characterization of Hepatozoon canis in dogs from urban and rural areas in Southeast Brazil

Research in Veterinary Science ◽

10.1016/j.rvsc.2014.06.015 ◽

2014 ◽

Vol 97 (2) ◽

pp. 325-328 ◽

Cited By ~ 21

Author(s):

R.L. de Miranda ◽

L.H. O'Dwyer ◽

J.R. de Castro ◽

B. Metzger ◽

A.S. Rubini ◽

...

Keyword(s):

Molecular Characterization ◽

Rural Areas ◽

Hepatozoon Canis ◽

Southeast Brazil ◽

Urban And Rural Areas

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Molecular characterization of porcine epidemic diarrhoea virus (PEDV) in Poland reveals the presence of swine enteric coronavirus (SeCoV) sequence in S gene

PLoS ONE ◽

10.1371/journal.pone.0258318 ◽

2021 ◽

Vol 16 (10) ◽

pp. e0258318

Author(s):

Marta Antas ◽

Monika Olech ◽

Anna Szczotka-Bochniarz

Keyword(s):

Phylogenetic Analysis ◽

Molecular Characterization ◽

Rna Virus ◽

Economic Losses ◽

High Morbidity ◽

Antigenic Analysis ◽

S Gene ◽

Porcine Epidemic Diarrhoea Virus ◽

Diarrhoea Virus

Porcine epidemic diarrhoea (PED) is a highly contagious enteric viral disease of pigs with a high morbidity and mortality rate, which ultimately results in huge economic losses in the pig production sector. The etiological agent of this disease is the porcine epidemic diarrhoea virus (PEDV) which is an enveloped, positive single-stranded RNA virus. The aim of this study was to perform molecular characterization of PEDV to identify the strains circulating in Poland. In this study, 662 faecal samples from 2015 to 2021 were tested with reverse transcription quantitative real-time PCR (RT-qPCR) and the results showed that 3.8% of the tested samples revealed a positive result for PEDV. A phylogenetic analysis of the complete genome and complete S gene sequences showed that Polish PEDV strains belonged to the G1b (S-INDEL) subgroup and were closely related to the European PEDV strains isolated from 2014 to 2019. Furthermore, RDP4 analysis revealed that the Polish PEDV strains harboured a recombinant fragment of ~400 nt in the 5’ end of S gene with PEDV and swine enteric coronavirus (SeCoV) being the major and minor parents, respectively. Antigenic analysis showed that the aa sequences of neutralizing epitopes were conserved among the Polish PEDV strains. Only one strain, #0100/5P, had a unique substitution in the COE epitope. However, Polish PEDV strains showed several substitutions, especially in the COE antigen, as compared to the classical strain CV777. To the best of our knowledge, this is the first report concerning the molecular characterization of porcine epidemic diarrhoea virus strains, as well as the first phylogenetic analysis for PEDV in Poland.

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Molecular characterization of the first human G15 rotavirus strain of zoonotic origin from the bovine species

Journal of General Virology ◽

10.1099/jgv.0.001581 ◽

2021 ◽

Vol 102 (4) ◽

Author(s):

Takeshi Tsugawa ◽

Yoshiki Fujii ◽

Yusuke Akane ◽

Saho Honjo ◽

Kenji Kondo ◽

...

Keyword(s):

Phylogenetic Analysis ◽

Molecular Characterization ◽

Human Infection ◽

Potential Reservoir ◽

Group A Rotaviruses ◽

Paediatric Outpatient ◽

Group A ◽

Human Infections ◽

Bovine Species

Group A rotaviruses (RVAs) infect a wide variety of mammalian and avian species. Animals act as a potential reservoir to RVA human infections by direct virion transmission or by contributing genes to reassortants. Here, we report the molecular characterization of a rare human RVA strain Ni17-46 with a genotype G15P[14], isolated in Japan in 2017 during rotavirus surveillance in a paediatric outpatient clinic. The genome constellation of this strain was G15-P[14]-I2-R2-C2-M2-A13-N2-T9-E2-H3. This is the first report of an RVA with G15 genotype in humans, and sequencing and phylogenetic analysis results suggest that human infection with this strain has zoonotic origin from the bovine species. Given the fact that this strain was isolated from a patient with gastroenteritis and dehydration symptoms, we must take into account the virulence of this strain in humans.

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Molecular characterization of small and medium segments of Crimean-Congo hemorrhagic fever virus in Turkey

Future Virology ◽

10.2217/fvl-2020-0003 ◽

2020 ◽

Vol 15 (4) ◽

pp. 247-254

Author(s):

Murat Karamese ◽

Erkan Ozmen ◽

Hakan Aydin ◽

Mehmet Ozkan Timurkan ◽

Mesud Fakirullahoglu

Keyword(s):

Infectious Disease ◽

Phylogenetic Analysis ◽

Phylogenetic Tree ◽

Molecular Characterization ◽

Hemorrhagic Fever ◽

Fever Virus ◽

Crimean Congo Hemorrhagic Fever ◽

Hemorrhagic Fever Virus ◽

Relationship Of

Aim: The objective was to investigate the genotypic relationship of S and M segments in Crimean-Congo hemorrhagic fever virus (CCHFV) by phylogenetic analysis in 25 patients from seven endemic cities in Turkey. Materials & methods: A total of 25 samples from patients with CCHF were included between 2012 and 2015. Phylogenetic tree analyses were inferred using MEGA version-6.0 and distances were calculated by Kimura’s 2-parameter. Results: Phylogenetic analysis showed that all isolated viruses (n = 25) were in the predicted clades such as clade V- Europe-1 regarding both S and M segments of the CCHFV. Conclusion: Further epidemiological, molecular and phylogenic studies should be performed in both reservoir animals/vectors and humans to determine the incidence of tick-borne infectious disease and to help to develop vaccines for prevention of the disease.

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Genomic Characterization and Phylogenetic Classification of Bovine Coronaviruses Through Whole Genome Sequence Analysis

Viruses ◽

10.3390/v12020183 ◽

2020 ◽

Vol 12 (2) ◽

pp. 183 ◽

Cited By ~ 11

Author(s):

Tohru Suzuki ◽

Yoshihiro Otake ◽

Satoko Uchimoto ◽

Ayako Hasebe ◽

Yusuke Goto

Keyword(s):

Phylogenetic Analysis ◽

Sequence Analysis ◽

Molecular Characterization ◽

Genome Sequence ◽

Whole Genome Sequence ◽

Whole Genome ◽

Wild Ruminant ◽

Genome Sequence Analysis ◽

Genomic Characterization

Bovine coronavirus (BCoV) is zoonotically transmissible among species, since BCoV-like viruses have been detected in wild ruminants and humans. BCoV causing enteric and respiratory disease is widespread in cattle farms worldwide; however, limited information is available regarding the molecular characterization of BCoV because of its large genome size, despite its significant economic impact. This study aimed to better understand the genomic characterization and evolutionary dynamics of BCoV via comparative sequence and phylogenetic analyses through whole genome sequence analysis using 67 BCoV isolates collected throughout Japan from 2006 to 2017. On comparing the genomic sequences of the 67 BCoVs, genetic variations were detected in 5 of 10 open reading frames (ORFs) in the BCoV genome. Phylogenetic analysis using whole genomes from the 67 Japanese BCoV isolates in addition to those from 16 reference BCoV strains, revealed the existence of two major genotypes (classical and US wild ruminant genotypes). All Japanese BCoV isolates originated from the US wild ruminant genotype, and they tended to form the same clusters based on the year and farm of collection, not the disease type. Phylogenetic trees on hemagglutinin-esterase protein (HE), spike glycoprotein (S), nucleocapsid protein (N) genes and ORF1 revealed clusters similar to that on whole genome, suggesting that the evolution of BCoVs may be closely associated with variations in these genes. Furthermore, phylogenetic analysis of BCoV S genes including those of European and Asian BCoVs and human enteric coronavirus along with the Japanese BCoVs revealed that BCoVs differentiated into two major types (European and American types). Moreover, the European and American types were divided into eleven and three genotypes, respectively. Our analysis also demonstrated that BCoVs with different genotypes periodically emerged and predominantly circulated within the country. These findings provide useful information to elucidate the detailed molecular characterization of BCoVs, which have spread worldwide. Further genomic analyses of BCoV are essential to deepen the understanding of the evolution of this virus.

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Molecular Characterization of Leishmania RNA virus 2 in Leishmania major from Uzbekistan

Genes ◽

10.3390/genes10100830 ◽

2019 ◽

Vol 10 (10) ◽

pp. 830 ◽

Cited By ~ 4

Author(s):

Yuliya Kleschenko ◽

Danyil Grybchuk ◽

Nadezhda S. Matveeva ◽

Diego H. Macedo ◽

Evgeny N. Ponirovsky ◽

...

Keyword(s):

Phylogenetic Analysis ◽

Cutaneous Leishmaniasis ◽

Molecular Characterization ◽

Leishmania Major ◽

Geographical Origin ◽

Rna Virus ◽

Viral Sequences

Here we report sequence and phylogenetic analysis of two new isolates of Leishmania RNA virus 2 (LRV2) found in Leishmania major isolated from human patients with cutaneous leishmaniasis in south Uzbekistan. These new virus-infected flagellates were isolated in the same region of Uzbekistan and the viral sequences differed by only nineteen SNPs, all except one being silent mutations. Therefore, we concluded that they belong to a single LRV2 species. New viruses are closely related to the LRV2-Lmj-ASKH documented in Turkmenistan in 1995, which is congruent with their shared host (L. major) and common geographical origin.

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Morphological and Genetic Evidence that the Cyanobacterium Lyngbya wollei (Farlow ex Gomont) Speziale and Dyck Encompasses at Least Two Species

Applied and Environmental Microbiology ◽

10.1128/aem.02645-07 ◽

2008 ◽

Vol 74 (12) ◽

pp. 3710-3717 ◽

Cited By ~ 16

Author(s):

Jennifer J. Joyner ◽

R. Wayne Litaker ◽

Hans W. Paerl

Keyword(s):

Phylogenetic Analysis ◽

Habitat Degradation ◽

Sequence Similarity ◽

Inorganic Nitrogen ◽

Small Subunit ◽

Morphological Data ◽

Rrna Gene ◽

Ssu Rrna ◽

Lyngbya Wollei ◽

Ssu Rrna Gene

ABSTRACT Dense blooms of the cyanobacterium Lyngbya wollei are increasingly responsible for declining water quality and habitat degradation in numerous springs, rivers, and reservoirs. This research represents the first molecular phylogenetic analysis of L. wollei in comparison with the traditional morphological characterization of this species. Specimens were collected from several springs in Florida and a reservoir in North Carolina. Segments of the small-subunit (SSU) rRNA and nifH genes were PCR amplified, cloned, and sequenced. The phylogenetic analysis of the SSU rRNA gene revealed sequences that fell into three distinct subclusters, each with >97% sequence similarity. These were designated operational taxonomic unit 1 (OTU1), OTU2, and OTU3. Similarly, the nifH sequences fell into three distinct subclusters named S1, S2, and S3. When either bulk samples or individual filaments were analyzed, we recovered OTU1 with S1, OTU2 with S2, and OTU3 with S3. The coherence between the three SSU rRNA gene and nifH subclusters was consistent with genetically distinct strains or species. Cells associated with subclusters OTU3 and S3 were significantly wider and longer than those associated with other subclusters. The combined molecular and morphological data indicate that the species commonly identified as L. wollei in the literature represents two or possibly more species. Springs containing OTU3 and S3 demonstrated lower ion concentrations than other collection sites. Geographical locations of Lyngbya subclusters did not correlate with residual dissolved inorganic nitrogen or phosphorus concentrations. This study emphasizes the need to complement traditional identification with molecular characterization to more definitively detect and characterize harmful cyanobacterial species or strains.

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