Morphological and molecular characterization of Trypanosoma copemani n. sp. (Trypanosomatidae) isolated from Gilbert's potoroo (Potorous gilbertii) and quokka (Setonix brachyurus)

Parasitology ◽  
2009 ◽  
Vol 136 (7) ◽  
pp. 783-792 ◽  
Author(s):  
J. M. AUSTEN ◽  
R. JEFFERIES ◽  
J. A. FRIEND ◽  
U. RYAN ◽  
P. ADAMS ◽  
...  
Keyword(s):  
Western Australia ◽  
Morphological Characteristics ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Bettongia Penicillata ◽  
Blood Smears ◽  
Common Wombat ◽  
Morphological And Molecular Characterization

SUMMARYLittle is known of the prevalence and life-cycle of trypanosomes in mammals native to Australia. Native Australian trypanosomes have previously been identified in marsupials in the eastern states of Australia, with one recent report in brush-tailed bettongs (Bettongia penicillata), or woylie in Western Australia in 2008. This study reports a novel Trypanosoma sp. identified in blood smears, from 7 critically endangered Gilbert's potoroos (Potorous gilbertii) and 3 quokkas (Setonix brachyurus) in Western Australia. Trypanosomes were successfully cultured in vitro and showed morphological characteristics similar to members of the subgenus Herpetosoma. Phylogenetic analysis of 18S rRNA gene sequences identified 2 different novel genotypes A and B that are closely related to trypanosomes previously isolated from a common wombat (Vombatus ursinus) in Victoria, Australia. The new species is proposed to be named Trypanosoma copemani n. sp.

LABORATORY DIAGNOSTIC OF HUMAN BABESIOSIS

2019 ◽  
Vol 64 (9) ◽  
pp. 560-564
Author(s):  
I. V. Kukina ◽  
O. P. Zelya ◽  
L. S. Karan
Keyword(s):  
Clinical Manifestations ◽  
Morphological Characteristics ◽  
Laboratory Diagnosis ◽  
18S Rrna Gene ◽  
Host Cells ◽  
Rrna Gene ◽  
Reference Centre ◽  
Blood Smears ◽  
Fatal Course ◽  
Human Babesiosis

Human babesiosis caused by parasitic protozoan Babesia spp. is sporadic zoonotic vector-borne infection. The course of babesiosis and prognosis depend on the type of pathogen and on the patient’s immunological status. Significance this disease is a severe, often fatal course with immunocompromissed patients resembling complicated falciparum malaria. In Europe to date, more than 50 cases of confirmed human babesiosis have been reported in most cases caused by Babesia divergens. Possible there are unrecognized cases. Pathogen is an obligate intraerythrocyte parasite of vertebrate animals. The organism is transmitted from animal to man through bite of Ixodidae tick. Asexual reproduction of the parasite occurs in a vertebrate host. The pathogenesis of babesiosis is caused by the destruction of host cells. Intensive haemolysis of red blood cells leads to the development of haemolytic anemia, haematuria, jaundice, and polyorgan failure may develop. The clinical manifestations of the disease are nonspecific. Detection of intraerythrocyte parasites in blood smears stained Gimsa-Romanovsky confirms the proposed diagnosis. Blood smears and some laboratory signs from fatal cases were analyzed in the Reference-centre of E. I. Martsinovskiĭ Institute. Original microphotographs B. divergens are shown. The main morphological forms of the parasite are shown. In addition to the well-known tetrades of parasites «Maltese Cross», for the first time, the parasites dividing into 6 interconnected trophozoites - “sextet” - were found. Originally, the invasion of Babesia in a normoblast is shown. An unusually high multiple invasion (14 parasites) of erythrocytes is noted. Because the patients, initially, were incorrectly diagnosed with malaria, the differential diagnosis of Babesia with Plasmodium is described step-by-step. It is important, since the treatment with antimalarial drugs is ineffective. Deviation laboratory signs are discussed. Complex morphological characteristics allowed us to speciated the parasites as B. divergens. DNA was detected in the sample with specific primers Bab di hsp70F/Bab di hsp70R and the probe Bab di hsp70P. The sequence demonstrated 99-100% and 98% similarity to the 18S rRNA gene fragment of B. divergence and Babesia venatorum, respectively. Molecular biological and serological methods of laboratory diagnosis of babesiosis are considered.

scholarly journals Molecular Characterization of Ciliate Diversity in Stream Biofilms

2008 ◽  
Vol 74 (6) ◽  
pp. 1740-1747 ◽  
Author(s):  
Andrew Dopheide ◽  
Gavin Lear ◽  
Rebecca Stott ◽  
Gillian Lewis
Keyword(s):  
18S Rrna ◽  
Pcr Primers ◽  
18S Rrna Gene ◽  
Sequence Diversity ◽  
Rrna Genes ◽  
Rrna Gene ◽  
Sequencing Analysis ◽  
Specific Pcr ◽  
Stream Biofilms

ABSTRACT Free-living protozoa are thought to be of fundamental importance in aquatic ecosystems, but there is limited understanding of their diversity and ecological role, particularly in surface-associated communities such as biofilms. Existing eukaryote-specific PCR primers were used to survey 18S rRNA gene sequence diversity in stream biofilms but poorly revealed protozoan diversity, demonstrating a need for protozoan-targeted primers. Group-specific PCR primers targeting 18S rRNA genes of the protozoan phylum Ciliophora were therefore designed and tested using DNA extracted from cultured protozoan isolates. The two most reliable primer combinations were applied to stream biofilm DNA, followed by cloning and sequencing analysis. Of 44 clones derived from primer set 384F/1147R, 86% were of probable ciliate origin, as were 25% of 44 clones detected by primer set 121F/1147R. A further 29% of 121F/1147R-detected clones matched sequences from the closely related phylum Apicomplexa. The highly ciliate-specific primer set 384F/1147R was subsequently used in PCRs on biofilm DNA from four streams exhibiting different levels of human impact, revealing differences in ciliate sequence diversity in samples from each site. Of a total of 240 clones, 73% were of probable ciliate origin; 54 different putative ciliate sequences were detected from throughout seven taxonomic ciliate classes. Sequences from Oligohymenophorea were most commonly detected in all samples, followed by either Spirotrichea or Phyllopharyngea. Restriction fragment length polymorphism profile-based analysis of clones suggested a potentially higher level of diversity than did sequencing. Nevertheless, newly designed PCR primers 384F/1147R were considered to provide an effective molecular basis for characterization of ciliate diversity in stream biofilms.

scholarly journals Detection of Protozoan Hosts for Legionella pneumophila in Engineered Water Systems by Using a Biofilm Batch Test

2010 ◽  
Vol 76 (21) ◽  
pp. 7144-7153 ◽  
Author(s):  
Rinske M. Valster ◽  
Bart A. Wullings ◽  
Dick van der Kooij
Keyword(s):  
Legionella Pneumophila ◽  
Biofilm Formation ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Batch Test ◽  
Free Living ◽  
Water Types ◽  
Operational Taxonomic Units ◽  
Hartmannella Vermiformis

ABSTRACT Legionella pneumophila proliferates in aquatic habitats within free-living protozoa, 17 species of which have been identified as hosts by using in vitro experiments. The present study aimed at identifying protozoan hosts for L. pneumophila by using a biofilm batch test (BBT). Samples (600 ml) collected from 21 engineered freshwater systems, with added polyethylene cylinders to promote biofilm formation, were inoculated with L. pneumophila and subsequently incubated at 37°C for 20 days. Growth of L. pneumophila was observed in 16 of 18 water types when the host protozoan Hartmannella vermiformis was added. Twelve of the tested water types supported growth of L. pneumophila or indigenous Legionella anisa without added H. vermiformis. In 12 of 19 BBT flasks H. vermiformis was indicated as a host, based on the ratio between maximum concentrations of L. pneumophila and H. vermiformis, determined with quantitative PCR (Q-PCR), and the composition of clone libraries of partial 18S rRNA gene fragments. Analyses of 609 eukaryotic clones from the BBTs revealed that 68 operational taxonomic units (OTUs) showed the highest similarity to free-living protozoa. Forty percent of the sequences clustering with protozoa showed ≥99.5% similarity to H. vermiformis. None of the other protozoa serving as hosts in in vitro studies were detected in the BBTs. In several tests with growth of L. pneumophila, the protozoa Diphylleia rotans, Echinamoeba thermarum, and Neoparamoeba sp. were identified as candidate hosts. In vitro studies are needed to confirm their role as hosts for L. pneumophila. Unidentified protozoa were implicated as hosts for uncultured Legionella spp. grown in BBT flasks at 15°C.

scholarly journals The New Human Babesia sp. FR1 Is a European Member of the Babesia sp. MO1 Clade

Pathogens ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 1433
Author(s):  
Claire Bonsergent ◽  
Marie-Charlotte de Carné ◽  
Nathalie de la Cotte ◽  
François Moussel ◽  
Véronique Perronne ◽  
...  
Keyword(s):  
Roe Deer ◽  
Phylogenetic Analyses ◽  
18S Rrna Gene ◽  
Natural Host ◽  
Etiological Agent ◽  
Rrna Gene ◽  
Separate Species ◽  
Apical Membrane Antigen 1 ◽  
Babesia Divergens

In Europe, Babesia divergens is responsible for most of the severe cases of human babesiosis. In the present study, we describe a case of babesiosis in a splenectomized patient in France and report a detailed molecular characterization of the etiological agent, named Babesia sp. FR1, as well as of closely related Babesia divergens, Babesia capreoli and Babesia sp. MO1-like parasites. The analysis of the conserved 18S rRNA gene was supplemented with the analysis of more discriminant markers involved in the red blood cell invasion process: rap-1a (rhoptry-associated-protein 1) and ama-1 (apical-membrane-antigen 1). The rap-1a and ama-1 phylogenetic analyses were congruent, placing Babesia sp. FR1, the new European etiological agent, in the American cluster of Babesia sp. MO1-like parasites. Based on two additional markers, our analysis confirms the clear separation of B. divergens and B. capreoli. Babesia sp. MO1-like parasites should also be considered as a separate species, with the rabbit as its natural host, differing from those of B. divergens (cattle) and B. capreoli (roe deer). The natural host of Babesia sp. FR1 remains to be discovered.

Naumovozyma baii sp. nov., an ascomycetous yeast species isolated from rotten wood in a tropical forest

2012 ◽  
Vol 62 (Pt_12) ◽  
pp. 3095-3098 ◽  
Author(s):  
Wan-Qiu Liu ◽  
Pei-Jie Han ◽  
Jun-Zhi Qiu ◽  
Qi-Ming Wang
Keyword(s):  
Gene Sequence ◽  
Yeast Species ◽  
Morphological Characteristics ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Internal Transcribed Spacer Region ◽  
Ascomycetous Yeast ◽  
Rotten Wood ◽  
D2 Domain ◽  
26S Rrna

Two strains isolated from rotten wood were included in the Saccharomyces group based on morphological characteristics. However, rRNA gene sequence analyses (including the 18S rRNA gene, 26S rRNA gene D1/D2 domain and internal transcribed spacer region) indicated that these two strains represent a novel species of Naumovozyma, for which the name Naumovozyma baii sp. nov. is proposed (type strain: BW 22T = CGMCC 2.04520T = CBS 12642T). The MycoBank number of the new species is MB800484.

scholarly journals Cryptosporidium spp. in Domestic Dogs: the “Dog” Genotype

2000 ◽  
Vol 66 (5) ◽  
pp. 2220-2223 ◽  
Author(s):  
Una M. Morgan ◽  
Lihua Xiao ◽  
Paul Monis ◽  
Abbie Fall ◽  
Peter J. Irwin ◽  
...  
Keyword(s):  
18S Rrna ◽  
18S Rrna Gene ◽  
Domestic Dogs ◽  
Heat Shock Gene ◽  
Valid Species ◽  
Rrna Gene ◽  
Phylogenetic Characterization ◽  
Cryptosporidium Spp ◽  
Shock Gene

ABSTRACT Genetic and phylogenetic characterization ofCryptosporidium isolates at two loci (18S rRNA gene and heat shock gene) from both Australian and United States dogs demonstrated that dog-derived Cryptosporidium isolates had a distinct genotype which is conserved across geographic areas. Phylogenetic analysis provided support for the idea that the “dog” genotype is, in fact, a valid species.

scholarly journals Genotypic Characterization of Torymus sinensis (Hymenoptera: Torymidae) After Its Introduction in Tuscany (Italy) for the Biological Control of Dryocosmus kuriphilus (Hymenoptera: Cynipidae)

2019 ◽  
Vol 19 (4) ◽  
Author(s):  
Ambra Viviani ◽  
Rodolfo Bernardi ◽  
Andrea Cavallini ◽  
Elisabetta Rossi
Keyword(s):  
Biological Control ◽  
18S Rrna ◽  
18S Rrna Gene ◽  
Its2 Region ◽  
Rrna Gene ◽  
Gall Wasp ◽  
Dryocosmus Kuriphilus ◽  
Torymus Sinensis ◽  
The World

Abstract Torymus sinensis Kamijo (Hymenoptera: Torymidae) is an alien parasitoid that is used in many areas of the world for biological control the Asian chestnut gall wasp, Dryocosmus kuriphilus Yasumatsu (Hymenoptera: Cynipidae). In Italy, this parasitoid was imported from Japan in 2003 and subsequently multiplied and released throughout the country. In this study, a phylogenetic investigation was carried out on insects from three different sites in northern Tuscany (Italy). Moreover, the possible hybridization between T. sinensis and some native Torymus species was evaluated. The conserved region 18S rRNA gene and the hypervariable ITS2 (Internal Transcribed Spacer 2) region of the ribosomal cistrone were selected as molecular markers. Sequencing the amplified products, after cloning, ruled out any hybridization between T. sinensis and the native Torymus species, and also confirmed the presence of two haplotypes for the Tuscan population of T. sinensis both for the region of the 18S rRNA gene as well as for the ITS2 region. These results confirm that the environmental impact of the alien parasitoid T. sinensis in the study site is acceptable, although an extensive and repeated monitoring would be desirable.

scholarly journals Fungal Colonization and Biodeterioration of Plasticized Polyvinyl Chloride

2000 ◽  
Vol 66 (8) ◽  
pp. 3194-3200 ◽  
Author(s):  
Jeremy S. Webb ◽  
Marianne Nixon ◽  
Ian M. Eastwood ◽  
Malcolm Greenhalgh ◽  
Geoffrey D. Robson ◽  
...  
Keyword(s):  
Bacterial Colonization ◽  
Morphological Characteristics ◽  
Sole Source ◽  
Dioctyl Phthalate ◽  
Microbial Colonization ◽  
Fungal Colonization ◽  
In Vitro Tests ◽  
Rrna Gene ◽  
Microbial Succession

ABSTRACT Significant substratum damage can occur when plasticized PVC (pPVC) is colonized by microorganisms. We investigated microbial colonization of pPVC in an in situ, longitudinal study. Pieces of pPVC containing the plasticizers dioctyl phthalate and dioctyl adipate (DOA) were exposed to the atmosphere for up to 2 years. Fungal and bacterial populations were quantified, and colonizing fungi were identified by rRNA gene sequencing and morphological characteristics.Aureobasidium pullulans was the principal colonizing fungus, establishing itself on the pPVC between 25 and 40 weeks of exposure. A group of yeasts and yeast-like fungi, includingRhodotorula aurantiaca and Kluyveromyces spp., established themselves on the pPVC much later (after 80 weeks of exposure). Numerically, these organisms dominated A. pullulans after 95 weeks, with a mean viable count ± standard error of 1,000 ± 200 yeast CFU cm−2, compared to 390 ± 50 A. pullulans CFU cm−2. No bacterial colonization was observed. We also used in vitro tests to characterize the deteriogenic properties of fungi isolated from the pPVC. All strains of A. pullulans tested could grow with the intact pPVC formulation as the sole source of carbon, degrade the plasticizer DOA, produce extracellular esterase, and cause weight loss of the substratum during growth in vitro. In contrast, several yeast isolates could not grow on pPVC or degrade DOA. These results suggest that microbial succession may occur during the colonization of pPVC and that A. pullulans is critical to the establishment of a microbial community on pPVC.

scholarly journals Genetic characterization of Strongyloides spp. from captive, semi-captive and wild Bornean orangutans (Pongo pygmaeus) in Central and East Kalimantan, Borneo, Indonesia

Parasitology ◽  
2011 ◽  
Vol 138 (11) ◽  
pp. 1417-1422 ◽  
Author(s):  
E. M. LABES ◽  
N. WIJAYANTI ◽  
P. DEPLAZES ◽  
A. MATHIS
Keyword(s):  
18S Rrna ◽  
Genetic Characterization ◽  
18S Rrna Gene ◽  
Rdna Locus ◽  
Rrna Gene ◽  
Fecal Material ◽  
East Kalimantan ◽  
The One ◽  
Bornean Orangutans

SUMMARYOrangutans (Pongo spp.), Asia's only great apes, are threatened in their survival due to habitat loss, hunting and infections. Nematodes of the genus Strongyloides may represent a severe cause of death in wild and captive individuals. In order to better understand which Strongyloides species/subspecies infect orangutans under different conditions, larvae were isolated from fecal material collected in Indonesia from 9 captive, 2 semi-captive and 9 wild individuals, 18 captive groups of Bornean orangutans and from 1 human working with wild orangutans. Genotyping was done at the genomic rDNA locus (part of the 18S rRNA gene and internal transcribed spacer 1, ITS1) by sequencing amplicons. Thirty isolates, including the one from the human, could be identified as S. fuelleborni fuelleborni with 18S rRNA gene identities of 98·5–100%, with a corresponding published sequence. The ITS1 sequences could be determined for 17 of these isolates revealing a huge variability and 2 main clusters without obvious pattern with regard to attributes of the hosts. The ITS1 amplicons of 2 isolates were cloned and sequenced, revealing considerable variability indicative of mixed infections. One isolate from a captive individual was identified as S. stercoralis (18S rRNA) and showed 99% identity (ITS1) with S. stercoralis sequences from geographically distinct locations and host species. The findings are significant with regard to the zoonotic nature of these parasites and might contribute to the conservation of remaining orangutan populations.

Illustration of Key Morphological Characteristics of Phytophthora cajani-Pathogen of Phytophthora Blight of Pigeonpea

2020 ◽  
Author(s):  
G. Jadesha ◽  
Mamta Sharma ◽  
P. Narayan Reddy
Keyword(s):  
Phase Contrast ◽  
Morphological Characteristics ◽  
Phytophthora Blight ◽  
Phytophthora Spp ◽  
Cropping Season ◽  
Contrast Microscope ◽  
Culture Characteristics ◽  
Stimulation Of

Background: Phytophthora cajani causing the Phytophthora blight (PB) disease of pigeonpea. The disease will rampant during excessive rainfall coupled with hot and humid weather during the cropping season. The present study on micro and macro morphological characteristics can contribute to the identification and specification of biology of Phytophthora spp. There are no detailed studies concerning the characterization of the P. cajani are available with this backdrop the present investigation was taken. Methods: Phytophthora cajani was isolated on V-8 PARP medium, whereas stimulation of zoospores and sporangia was done using the diluted tomato juice broth. Micro and macro morphological characteristics of P. cajani were studied using micrometry and Olympus CX41 phase-contrast microscope. Result: The pathogen was homothallic with amphigynous antheridium and oogonium and able to produce oospore in vitro. Sporangium was nonpapillate, noncaducous, oviod-obpyriform shape. Further, the macro morphological characteristics like mycelial radial growth and colony type were studied. The colony characteristics were dull white, flat and rosette pattern. Other culture characteristics like optimum temperature and RH were mostly consistent with those reported former.

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