Surveillance of Giardia and Cryptosporidium in sewage from an urban area in Brazil

Abstract Cryptosporidium and Giardia are protozoan parasites that cause diarrhea in humans and animals. Molecular characterization of these pathogens in sewage may provide insight on their occurrence and prevalence in Brazil. This study aimed to investigate the presence of Giardia and Cryptosporidium in raw and treated sewage from Londrina, Paraná, Brazil. Samples were collected every two weeks during a year. Samples were concentrated, then DNA was extracted and subjected to a nested PCR targeting the Giardia 18S rRNA gene and the Cryptosporidium 18S rRNA gene. Species of Cryptosporidium were characterized by restriction fragment length polymorphism (RFLP). All raw sewage and 76% of the treated sewage were positive for Giardia; 84% of raw sewage samples and 8% of treated sewage were positive for Cryptosporidium. C. muris, C. hominis, C. baileyi, C. parvum and C. suis were detected in 100%, 19%, 9%, 9% and 4% of raw sewage, respectively. C. muris was the only species found in treated sewage. Multiple species of Cryptosporidium were present in 19.04% of the raw sewage. Treated sewage water can pose a threat to human health. The speciation of Cryptosporidium revealed the presence of non-common zoonotic species as C. suis and C. muris.

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

Journal of Insect Science ◽

10.1093/jisesa/iez080 ◽

2019 ◽

Vol 19 (4) ◽

Cited By ~ 1

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.

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

Parasitology ◽

10.1017/s0031182011001284 ◽

2011 ◽

Vol 138 (11) ◽

pp. 1417-1422 ◽

Cited By ~ 17

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.

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Characterization of Hydrocortisone Biometabolites and 18S rRNA Gene in Chlamydomonas reinhardtii Cultures

Molecules ◽

10.3390/molecules13102416 ◽

2008 ◽

Vol 13 (10) ◽

pp. 2416-2425 ◽

Cited By ~ 12

Author(s):

Younes Ghasemi ◽

Sara Rasoul-Amini ◽

Mohammad Morowvat ◽

Mohammad Raee ◽

Mohammad Ghoshoon ◽

...

Keyword(s):

Chlamydomonas Reinhardtii ◽

18S Rrna ◽

18S Rrna Gene ◽

Rrna Gene

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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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Rapid Detection of Equine Piroplasms Using Multiplex PCR and First Genetic Characterization of Theileria haneyi in Egypt

Pathogens ◽

10.3390/pathogens10111414 ◽

2021 ◽

Vol 10 (11) ◽

pp. 1414

Author(s):

Bassma S. M. Elsawy ◽

Ahmed M. Nassar ◽

Heba F. Alzan ◽

Raksha V. Bhoora ◽

Sezayi Ozubek ◽

...

Keyword(s):

Multiplex Pcr ◽

Genetic Characterization ◽

Simultaneous Detection ◽

18S Rrna Gene ◽

Rrna Gene ◽

Theileria Equi ◽

Babesia Caballi ◽

Pcr Targeting ◽

First Time

Equine Piroplasmosis (EP) is an infectious disease caused by the hemoprotozoan parasites Theileria equi, Babesia caballi, and the recently identified species T. haneyi. Hereby, we used a multiplex PCR (mPCR) targeting the 18S rRNA gene of T. equi and B. caballi for the simultaneous detection of EP in Egyptian equids and examined the presence of T. haneyi infections in Egypt. Blood samples from 155 equids (79 horses and 76 donkeys) collected from different governorates of Egypt were examined by mPCR and PCR targeting T. hayeni. The mPCR method revealed a prevalence of T. equi of 20.3% in horses and of 13.1% in donkeys and a prevalence of B. caballi of 1.2% in horses. B. caballi was not detected in donkeys in the current study. The mPCR method also detected coinfections with both species (2.5% and 1.3% in horses and donkeys, respectively). Additionally, we report the presence of T. haneyi in Egypt for the first time in 53.1% of the horse and 38.1% of the donkey tested samples. Coinfection with T. haneyi and T. equi was found in 13.5% of the samples, while infection with the three EP species was found in 1.9% of the samples.

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