scholarly journals Cryptosporidium spp. in wild murids (Rodentia) from Corsica, France

2021 ◽  
Author(s):  
Katherine García-Livia ◽  
Ángela Fernández-Álvarez ◽  
Carlos Feliu ◽  
Jordi Miquel ◽  
Yann Quilichini ◽  
...  
Keyword(s):  
Rattus Rattus ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Mus Musculus Domesticus ◽  
Protozoan Parasites ◽  
Wild Rodents ◽  
Genotype Iii ◽  
Mediterranean Island ◽  
Murine Rodents ◽  
Vertebrate Hosts

AbstractCryptosporidium spp. are worldwide protozoan parasites that can affect to a broad range of vertebrate hosts, including rodents. In the island of Corsica (France), there are no previous data about these protozoa infecting wild rodents. To estimate the distribution and occurrence, a total of 117 wild murine rodents of the species Rattus rattus (84), Mus musculus domesticus (21), Apodemus sylvaticus (11), and Rattus norvegicus (1) were captured in 24 different biotopes. Fecal samples were screened for Cryptosporidium spp. by nested PCR to amplify an 830 bp fragment of the 18S rRNA gene. As general occurrence, 15.4% of the rodents analyzed were positive for Cryptosporidium spp., being detected widely distributed along the island in R. rattus (17.6%) and M. m. domesticus (14.3%). Cryptosporidium viatorum, Cryptosporidium sp. rat genotype II, and Cryptosporidium sp. rat genotype III were successfully identified in R. rattus. The results herein reported provide the first data on Cryptosporidium spp. in wild murine species from a Mediterranean island and constitute the first report of the zoonotic species C. viatorum in R. rattus. Although a low occurrence of Cryptosporidium spp. in murids was obtained and only in one animal the zoonotic species C. viatorum was identified, our results highlight that wild murine rodents from Corsica could mediate in the maintenance and transmission of this protozoan to the environment and other hosts including humans and animals. Further studies are required to better understand the epidemiology of Cryptosporidium spp. in wild rodents from Corsica and their possible public health repercussions.

scholarly journals Apicomplexa primers amplify Proteromonas (Stramenopiles, Slopalinida, Proteromonadidae) in tissue and blood samples from lizards

2012 ◽  
Vol 57 (4) ◽  
Author(s):  
João Maia ◽  
Elena Gómez-Díaz ◽  
D. Harris
Keyword(s):  
Cross Validation ◽  
Molecular Data ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Molecular Screening ◽  
Protozoan Parasites ◽  
Apicomplexan Parasites ◽  
Wide Range ◽  
Prevalence Estimates ◽  
Vertebrate Hosts

AbstractMicroscopy has traditionally been the most common method in parasitological studies, but in recent years molecular screening has become increasingly frequent to detect protozoan parasites in a wide range of vertebrate hosts and vectors. During routine molecular screening of apicomplexan parasites in reptiles using the 18S rRNA gene, we have amplified and sequenced Proteromonas parasites from three lizard hosts (less than 1% prevalence). We conducted phylogenetic analysis to confirm the taxonomic position and infer their relationships with other stramenopiles. Although our phylogeny is limited due to scarcity of molecular data on these protists, our results confirm they are closely related to Proteromonas lacertae. Our findings show that unexpected parasites can be amplified from host samples (blood and tissue) using general procedures to detect hemoparasites, and stress that positive PCR amplifications alone should not be considered as definitive proof of infection by particular parasites. Further validation by sequence confirmation and thorough phylogenetic assessment will not only avoid false positives and biased prevalence estimates but also provide valuable information on the biodiversity and phylogenetic relationships of other parasitic organisms. More generally, our results illustrate the perils of general diagnosis protocols in parasitological studies and the need of cross-validation procedures.

scholarly journals Molecular Characterization ofCryptosporidiumspp. in Wild Rodents of Southwestern Iran Using 18s rRNA Gene Nested-PCR-RFLP and Sequencing Techniques

2016 ◽  
Vol 2016 ◽  
pp. 1-6 ◽  
Author(s):  
Jasem Saki ◽  
Masoud Foroutan-Rad ◽  
Reza Asadpouri
Keyword(s):  
Molecular Characterization ◽  
Nested Pcr ◽  
18S Rrna ◽  
Restriction Enzymes ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Wild Rodents ◽  
Nested Polymerase Chain Reaction ◽  
Pcr Rflp ◽  
Southwest Of Iran

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.

Molecular Survey of Babesia microti (Aconoidasida: Piroplasmida) in Wild Rodents in Turkey

2019 ◽  
Vol 56 (6) ◽  
pp. 1605-1609 ◽  
Author(s):  
Selma Usluca ◽  
Bekir Celebi ◽  
Djursun Karasartova ◽  
A Semra Gureser ◽  
Ferhat Matur ◽  
...  
Keyword(s):  
Sequence Data ◽  
18S Rrna Gene ◽  
Babesia Microti ◽  
Myodes Glareolus ◽  
Rrna Gene ◽  
Wild Rodents ◽  
Sarcocystis Spp ◽  
Zoonotic Parasite ◽  
Reservoir Hosts ◽  
Babesia Spp

Abstract Babesia microti (Aconoidasida: Piroplasmida) (Franca, 1910) is an important tick-borne zoonotic parasite with rodents serving as reservoir hosts. In the present study, 536 rodents were captured from Burdur, Bartin, Giresun, and Yozgat provinces of Turkey between the years 2010 and 2012, and blood samples were examined for the presence of Babesia spp. using conventional PCR which targeted the 18S rRNA gene. The sequence analysis of PCR amplicons was tested for B. microti as well as for Hepatozoon spp., and Sarcocystis spp. Overall, 5.8% of the rodents were positive for B. microti: 41% in Myodes glareolus, 7.7% in Chionomys roberti, and 2% in Apodemus spp., whereas no Babesia DNA was detected in Mus macedonicus and Microtus spp. Six rodents were positive for Hepatozoon spp. and one rodent was positive for Sarcocystis spp. Overall, 14.9 and 4.5% of rodents captured from Bartin and Giresun provinces, respectively, were PCR positive for B. microti, whereas none of rodents captured in Burdur and Yozgat were positive for Babesia spp. The sequence data of B. microti from rodents revealed that all sequences belonged to the zoonotic genotype. Sequences of B. microti obtained from rodents of the Bartin province were genotypically closer to European isolates, whereas those obtained from rodents of the Giresun province were closer to Russian and Mongolian isolates.

scholarly journals Molecular prevalence of Theileria infections in cattle in Yanbian, north-eastern China

Parasite ◽  
2020 ◽  
Vol 27 ◽  
pp. 19
Author(s):  
Lijun Jia ◽  
Shaowei Zhao ◽  
Suzhu Xie ◽  
Hang Li ◽  
Hao Wang ◽  
...  
Keyword(s):  
Eastern China ◽  
Epidemiological Data ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Protozoan Parasites ◽  
Total Prevalence ◽  
North Eastern ◽  
Bovine Theileriosis ◽  
And Control ◽  
Bovine Erythrocytes

Bovine Theileria are tick-borne protozoan parasites that invade bovine erythrocytes and lymphocytes. Three main bovine Theileria species have been identified in China: T. orientalis, T. sinensis, and T. annulata. To examine the prevalence of bovine theileriosis in Yanbian, a total of 584 bovine blood samples were collected from five localities from 2017 to 2019 and analyzed by PCR. Six pairs of oligonucleotide primers directed against the 18S rRNA gene of Theileria spp., Tams-1 gene of T. annulata, MPSP gene of T. orientalis, and T. sinensis, were used to detect these parasites. A sequence analysis of the amplified genes confirmed that the Theileria species were T. orientalis and T. sinensis, without T. annulata. The overall prevalence of Theileria in cattle was 42.81% (250/584). Out of the 584 samples, 159 (27.23%) and 157 (26.88%) were positive for T. sinensis and T. orientalis, respectively, and the mixed infection rate was 11.30% (66/584). The total prevalence of bovine Theileria species in Helong, Hunchun, Longjing, Yanji, and Dunhua was 66.28%, 49.68%, 23.81%, 28.15%, and 0%, respectively. These results provide epidemiological data for the prevention and control of bovine Theileria species in Yanbian, China.

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

2019 ◽  
Vol 28 (2) ◽  
pp. 291-297 ◽  
Author(s):  
Felippe Danyel Cardoso Martins ◽  
Winni Alves Ladeia ◽  
Roberta dos Santos Toledo ◽  
João Luis Garcia ◽  
Italmar Teodorico Navarro ◽  
...  
Keyword(s):  
18S Rrna ◽  
Fragment Length Polymorphism ◽  
Sewage Water ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Protozoan Parasites ◽  
Treated Sewage ◽  
Multiple Species ◽  
Pcr Targeting

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.

scholarly journals Molecular Detection ofTheileriaspp. in Livestock on Five Caribbean Islands

2015 ◽  
Vol 2015 ◽  
pp. 1-8 ◽  
Author(s):  
Jilei Zhang ◽  
Patrick Kelly ◽  
Jing Li ◽  
Chuanling Xu ◽  
Chengming Wang
Keyword(s):  
Molecular Detection ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Limited Information ◽  
Protozoan Parasites ◽  
Sheep And Goats ◽  
Caribbean Islands ◽  
Related Organism ◽  
Wide Range ◽  
The Caribbean

Theileriaspp. are tick-transmitted, intracellular apicomplexan protozoan parasites infecting a wide range of animals. As there is very limited information on the prevalence ofTheileriaspp. in the Caribbean we used the recently described genus-specific pan-TheileriaFRET-qPCR to identify infected animals in the region and a standard 18S rRNA gene PCR and sequencing to determine the species involved. We foundTheileriaspp. in 9% of the convenience samples of animals (n=752) studied from five Caribbean islands. Donkeys (20.0%: 5/25) were most commonly infected, followed by sheep (17.4%, 25/144), cattle (6.8%; 22/325), goats (5.0%; 12/238), and horses (5.0%; 1/20). Six species ofTheileriawere identified:T.equi(donkeys, cattle, goats, and sheep),Theileriasp. OT3 (sheep and goats),Theileriasp. NG-2013a (cattle),Theileriasp. YW-2014 (donkeys),Theileriasp. B15a (goats), andBabesia vulpesor a closely related organism (sheep and goats). OnlyT. equihas been previously reported in the Caribbean. Our findings expand the known host ranges ofTheileriaspp. and the known distribution of the organisms around the world.

Theileria ovis (Piroplasmida: Theileriidae) Detected in Melophagus ovinus (Diptera: Hippoboscoidea) and Ornithodoros lahorensis (Ixodida: Argasidae) Removed From Sheep in Xinjiang, China

2019 ◽  
Vol 57 (2) ◽  
pp. 631-635 ◽  
Author(s):  
Li Zhao ◽  
Jinling Wang ◽  
Yulin Ding ◽  
Kairui Li ◽  
Bo He ◽  
...  
Keyword(s):  
18S Rrna ◽  
18S Rrna Gene ◽  
Economic Losses ◽  
Rrna Gene ◽  
Protozoan Parasites ◽  
Autonomous Region ◽  
Apicomplexan Parasites ◽  
Wide Range ◽  
Melophagus Ovinus ◽  
Theileria Spp

Abstract Theileria spp. are tick-transmitted, intracellular apicomplexan protozoan parasites that infect a wide range of animals and, as such, can cause significant economic losses. The aim of the present study was to detect and analyze apicomplexan parasites from two different ectoparasites that were collected from Xinjiang Uygur Autonomous Region, China. The PCR-based detection of 18S rRNA indicated that Ornithodoros lahorensis specimens from Kashgar, Xinjiang, and Aksu were positive for Theileria spp., as were Melophagus ovinus specimens from Aksu. Meanwhile, phylogenetic analysis, based on the 18S rRNA gene sequences, revealed that the four amplified Theileria sequences could be attributed to T. ovis. To the best of our knowledge, this is the first study to report the detection of T. ovis DNA in M. ovinus and the first molecular identification study to confirm the detection of T. ovis in O. lahorensis in China. Accordingly, the present study extends the known distribution of T. ovis.

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.

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