Molecular characterization of Cryptosporidium xiaoi in goat kids in Bangladesh by nested PCR amplification of 18S rRNA gene

Background. Rodents could act as reservoir forCryptosporidiumspp. speciallyC. parvum, a zoonotic agent responsible for human infections. Since there is no information aboutCryptosporidiuminfection in rodents of Ahvaz city, southwest of Iran, hence, this survey was performed to determine the prevalence and molecular characterization ofCryptosporidiumspp. in this region.Materials and Methods. One hundred rodents were trapped from different regions of Ahvaz city. Intestine contents and fecal specimens of rodents were studied using both microscopy examination to identify oocyst and nested-polymerase chain reaction (PCR) technique for 18s rRNA gene detection. Eventually restriction fragment length polymorphism (RFLP) method usingSspIandVspIrestriction enzymes was carried out to genotype the species and then obtained results were sequenced.Results. Three out of 100 samples were diagnosed as positive and overall prevalence ofCryptosporidiumspp. was 3% using both modified Ziehl-Neelsen staining under light microscope and nested-PCR (830 bp) methods. Afterwards, PCR-RFLP was performed on positive samples andC. parvumpattern was identified. Finally PCR-RFLP findings were sequenced and presence ofC. parvumwas confirmed again.Conclusions. Our study showed rodents could be potential reservoir forC. parvum. So an integrated program for control and combat with them should be adopted and continued.

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Molecular Characterization of Cryptosporidium Isolates from Humans and Animals in Iran

Applied and Environmental Microbiology ◽

10.1128/aem.00964-06 ◽

2006 ◽

Vol 73 (3) ◽

pp. 1033-1035 ◽

Cited By ~ 32

Author(s):

Ahmad Reza Meamar ◽

Karine Guyot ◽

Gabriela Certad ◽

Eduardo Dei-Cas ◽

Mino Mohraz ◽

...

Keyword(s):

Molecular Characterization ◽

18S Rrna ◽

18S Rrna Gene ◽

Rrna Gene ◽

Predominant Species ◽

Animal Hosts

ABSTRACT Isolates of Cryptosporidium spp. from human and animal hosts in Iran were characterized on the basis of both the 18S rRNA gene and the Laxer locus. Three Cryptosporidium species, C. hominis, C. parvum, and C. meleagridis, were recognized, and zoonotically transmitted C. parvum was the predominant species found in humans.

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Molecular Characterization of Cryptosporidium spp. in Wild Rats of Tehran, Iran Using 18s rRNA Gene and PCR_RFLP Method

Jundishapur Journal of Microbiology ◽

10.5812/jjm.3580 ◽

2012 ◽

Vol 5 (3) ◽

pp. 486-490 ◽

Cited By ~ 8

Author(s):

Fares Bahrami ◽

Javid Sadraei ◽

Mehdi Frozandeh

Keyword(s):

Molecular Characterization ◽

18S Rrna ◽

18S Rrna Gene ◽

Rrna Gene ◽

Cryptosporidium Spp ◽

Wild Rats

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PP-185 Infection determination and molecular characterization of Cryptosporidium types in wild rats of Tehran for 18s rRNA gene with PCR-RFLP method

International Journal of Infectious Diseases ◽

10.1016/s1201-9712(11)60335-4 ◽

2011 ◽

Vol 15 ◽

pp. S96

Author(s):

B. Fares ◽

J. Sadraei

Keyword(s):

Molecular Characterization ◽

18S Rrna ◽

18S Rrna Gene ◽

Rrna Gene ◽

Pcr Rflp ◽

Wild Rats

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Molecular characterization of Cryptosporidium isolates from human and bovine using 18s rRNA gene in Shahriar county of Tehran, Iran

Parasitology Research ◽

10.1007/s00436-008-1008-2 ◽

2008 ◽

Vol 103 (2) ◽

pp. 467-472 ◽

Cited By ~ 21

Author(s):

Majid Pirestani ◽

Javid Sadraei ◽

Abdolhossein Dalimi asl ◽

Mahdi Zavvar ◽

Hossein Vaeznia

Keyword(s):

Molecular Characterization ◽

18S Rrna ◽

18S Rrna Gene ◽

Rrna Gene

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Molecular Characterization of Ciliate Diversity in Stream Biofilms

Applied and Environmental Microbiology ◽

10.1128/aem.01438-07 ◽

2008 ◽

Vol 74 (6) ◽

pp. 1740-1747 ◽

Cited By ~ 52

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.

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Crenosoma striatum in lungs of European hedgehogs (Erinaceus europeus) from Portugal

Helminthologia ◽

10.2478/helm-2020-0020 ◽

2020 ◽

Vol 57 (2) ◽

pp. 179-184

Author(s):

P. F. Barradas ◽

A. R. Flores ◽

T. L. Mateus ◽

F. Carvalho ◽

F. Gärtner ◽

...

Keyword(s):

Molecular Characterization ◽

18S Rrna ◽

Population Sample ◽

Genetic Data ◽

Rrna Genes ◽

12S Rrna ◽

Rrna Gene ◽

Mitochondrial 12S Rrna ◽

18S Rrna Genes

SummaryCrenosoma striatum is a host-specifi c metastrongiloid nematode causing respiratory tract disease in hedgehogs (Erinaceus europaeus). Since few studies have reported C. striatum in hedgehogs and little genetic data is available concerning this lungworm, this study aimed to determine the occurrence of C. striatum in a population sample of hedgehogs from Portugal, additionally providing morphological, histological and molecular data. From 2017 to 2018 a survey of infection was carried out in 11 necropsied hedgehogs. Worms were extracted from fresh lung tissues and microscopically evaluated. Molecular characterization of partial mitochondrial (12S rRNA) and nuclear (18S rRNA) genes was performed. The presence of lungworms in pulmonary tissues of five hedgehogs (45.5%) was detected. Morphological and histopathological analyses evidenced adult forms of nematodes consistent with C. striatum. Molecular characterization of 18S rRNA genes confirmed the classifi cation as C. striatum. Also, novel genetic data characterizing the mitochondrial (12S rRNA) gene of C. striatum is presented.This is the first report of C. striatum infection in hedgehogs of Portugal. The findings here reported provide new insights regarding the geographic distribution and the molecular identification of this lungworm species.

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

Applied and Environmental Microbiology ◽

10.1128/aem.66.5.2220-2223.2000 ◽

2000 ◽

Vol 66 (5) ◽

pp. 2220-2223 ◽

Cited By ~ 38

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.

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Genomic Profiling for Piroplasms in Feeding Ixodid Ticks in the Eastern Cape, South Africa

Pathogens ◽

10.3390/pathogens9121061 ◽

2020 ◽

Vol 9 (12) ◽

pp. 1061

Author(s):

Olusesan Adeyemi Adelabu ◽

Benson Chuks Iweriebor ◽

Anthony Ifeanyi Okoh ◽

Larry Chikwelu Obi

Keyword(s):

South Africa ◽

Nested Pcr ◽

18S Rrna ◽

18S Rrna Gene ◽

Rhipicephalus Appendiculatus ◽

Ixodid Ticks ◽

Farm Animals ◽

Homology Search ◽

Rrna Gene ◽

Eastern Cape

Importation of tick-infected animals and the uncontrollable migration of birds and wild animals across borders can lead to geographical expansion and redistribution of ticks and pathogen vectors, thus leading to the emergence and re-emergence of tick-borne diseases in humans and animals. Comparatively, little is known about the occurrence of piroplasms in ixodid ticks in the Eastern Cape, South Africa, thus necessitating this study, which is aimed at detecting piroplasms (Theileria and Babesia) from feeding tick samples collected from cattle, sheep, and goats in selected sites in the Eastern Cape, South Africa. A total of 1200 feeding ixodid ticks collected from farm animals at selected homesteads were first subjected to molecular identification using mitochondrial 12S ribosomal RNA (rRNA) gene by PCR and were further tested for the presence of piroplasms through amplification of the 18S rRNA gene via nested-PCR followed by sequencing of the PCR products. The results indicated that 853 (71.1%) corresponded to the genus Rhipicephalus, 335 (27.9%) corresponded to genus Amblyomma, and 12 (1%) corresponded to genus Haemaphysalis. Amblyomma hebraeum and Rhipicephalus appendiculatus were the most common identified ticks from this study. The 18S rRNA nested-PCR revealed that 44 (3.7%) samples were confirmed positive for Theileria. A homology search for the generated sequences revealed a high percentage identity of 98–98.9% similarity to T. buffeli, T. orientalis, and T. sergenti in the GenBank. Based on the results obtained herein, we conclude that there is a big diversity of Theileria species; therefore, we suggest that this research should cover more geographical areas in order to reveal the true prevalence of this pathogen in the studied area because this will be a great step in the possible prevention of an outbreak that could have devastating effects on livestock production and human health in both the studied areas and South Africa at large.

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