Molecular Identification and Genotyping of Babesia canis in Dogs from Mesh-kin Shahr County, Northwestern Iran

Background: Canine babesiosis is one of the mainly worldwide-distributed tick-borne haemoprotozoan parasitic dis-eases in dogs. Methods: A total of 43 blood samples were randomly collected from naturally infected dogs in seven villages from different geographical areas of Meshkin Shahr, Ardabil Province, Iran. The presence of Babesia species detected with standard methods including parasitological and gene sequencing techniques targeting the 18S rRNA gene. Results: Our results revealed that four dogs 9.3% (4/43) including one female and three male dogs were infected with Babesia. All four Babesia-infected dogs were confirmed B. canis by the molecular-based method. Sequence alignments comparison of the B. canis genotypes A and B, it was revealed that all B. canis isolates belonged to genotype B. Conclusion: This study provides essential data for subsequently define the critical importance of the molecular studies in management and prevention of the canine babesiosis in Iran.

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Analytical validation of a real-time hydrolysis probe PCR assay for quantifying Plasmodium falciparum parasites in experimentally infected human adults

Malaria Journal ◽

10.1186/s12936-021-03717-y ◽

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.

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Eukaryotic Community in UASB Reactor Treating Domestic Sewage Based on 18S rRNA Gene Sequencing

Lecture Notes in Civil Engineering - Frontiers in Wastewater Treatment and Modelling ◽

10.1007/978-3-319-58421-8_34 ◽

2017 ◽

pp. 218-224 ◽

Cited By ~ 2

Author(s):

Y. Hirakata ◽

M. Hatamoto ◽

M. Oshiki ◽

N. Araki ◽

T. Yamaguchi

Keyword(s):

18S Rrna ◽

Gene Sequencing ◽

18S Rrna Gene ◽

Domestic Sewage ◽

Uasb Reactor ◽

Rrna Gene ◽

Eukaryotic Community ◽

Rrna Gene Sequencing

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Ecological assessment of phytoplankton community via microscopic method and 18S rRNA gene sequencing in Pearl River Estuary

E3S Web of Conferences ◽

10.1051/e3sconf/201913101043 ◽

2019 ◽

Vol 131 ◽

pp. 01043 ◽

Cited By ~ 1

Author(s):

Bo Peng ◽

Yuannan Wang ◽

Hengjun Zhang ◽

Chen Chen ◽

Hailin Luo ◽

...

Keyword(s):

18S Rrna ◽

Phytoplankton Community ◽

Diversity Index ◽

River Estuary ◽

Gene Sequencing ◽

Pearl River Estuary ◽

Ecological Assessment ◽

18S Rrna Gene ◽

Rrna Gene ◽

Rrna Gene Sequencing

Monitoring phytoplankton community underpins our understanding of water quality and ecological functions. In this study, we approached phytoplankton abundance, community composition, and diversity by both microscopy and 18S rRNA gene sequencing. Environmental variances influencing the phytoplankton were evaluated as well. There were 6 phyla and 62 species identified by microscopy, and the diversity index Shannon-Wiener and evenness index Pielou index indicated phytoplankton community had high diversity; however, the high density of dominance genus suggested that our research region had potential red tide effects. The canonical correspondence analysis illustrated that suspended solids, phosphate and temperature were three major factors that affected the distribution and components of phytoplankton community. The DNA barcoding sequencing of 18S rRNA gene supported the main results via microscopic methods while providing more identified community components, which implied that 18S rRNA gene sequencing can be used as a supplemental method for fast ecological assessment of phytoplankton community.

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Association between diet and rumen microbiota in wild roe deer

FEMS Microbiology Letters ◽

10.1093/femsle/fnz060 ◽

2019 ◽

Vol 366 (6) ◽

Cited By ~ 1

Author(s):

Robert Wilson ◽

Kjartan Østbye ◽

Inga Leena Angell ◽

Knut Rudi

Keyword(s):

18S Rrna ◽

Roe Deer ◽

Gene Sequencing ◽

16S Rrna Gene Sequencing ◽

18S Rrna Gene ◽

Rrna Gene ◽

Rumen Microbiota ◽

Dietary Components ◽

Rrna Gene Sequencing ◽

Insight Into

ABSTRACT The association between diet and the rumen microbiota for wild animals remains largely unexplored. Here, we explored this association using a combination of 16S rRNA gene sequencing to determine the prokaryote microbiota and 18S rRNA gene sequencing to determine the dietary components for wild roe deer. These analyses revealed a wide diversity of dietary components, with over-representation of Bacteroidetes for the diet-correlating bacteria. Ruminococcus, on the other hand, dominated the stable diet-independent part of the microbiota. Taken together, the combination of 16S and 18S rRNA gene analyses provide novel insight into rumen microbiota ecology.

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Investigations on First Confirmed Outbreak of Ovine Theileriosis (Theileria luwenshuni) from Maharashtra State, India

Indian Journal of Animal Research ◽

10.18805/ijar.b-4199 ◽

2020 ◽

Author(s):

V.S. Dhaygude ◽

K. Kundu ◽

B.P. Kamdi ◽

U.R. Bagal ◽

S.B. Bhosale ◽

...

Keyword(s):

18S Rrna ◽

Clinical Signs ◽

Specific Treatment ◽

Small Ruminants ◽

18S Rrna Gene ◽

Pcr Detection ◽

Nucleotide Sequencing ◽

Rrna Gene ◽

Blood Samples ◽

Theileria Luwenshuni

Background: Clinical theileriosis of small ruminants is tick-borne disease caused by Theileria lestoquardi, Theileria uilenbergi and Theileria luwenshuni. Theileria annulata, the causative agent of bovine tropical theileriosis in cattle, can also infect sheep but does not cause any significant illness. It is one of the economically important diseases. There are no reports of ovine clinical theileriosis from Maharashtra state and there is paucity of information on its epidemiology. This paper reports first confirmed outbreak of ovine theileriosis based on clinical signs, microscopic examination, PCR and sequencing in the Maharashtra State of India. Methods: Whole blood samples from 22 ailing sheep were collected and subjected to hematological examination. Blood smears stained with Leishman’s stain were examined under 100X objective of the microscope. The blood samples from sheep found positive by microscopic method were subjected to PCR detection of 18S rRNA gene of hemoprotozoa and then for nucleotide sequencing and sequence analysis.Conclusion: Samples from 14 out of 22 sheep were found positive for piroplasms of Theileria spp by light microscopy. All positive samples were further confirmed by PCR detection of 18S rRNA gene of hemoprotozoa. PCR amplification yielded expected product of 1750 bp for all samples. BLAST and phylogenetic analysis of one sample revealed high sequence homology with T. luwenshuni reported from India and other countries. Characteristic clinical signs like fever, progressive anaemia, laboured breathing, lymphadenopathy, debility and non-responsiveness to antibiotic therapy were recorded. The animals responded to specific treatment against theileriosis. It is the first ever confirmed report of ovine theileriosis in Maharashtra state of India and hence reported.

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Diversity and distribution of nematodes associated with terrestrial slugs in the Western Cape Province of South Africa

Journal of Helminthology ◽

10.1017/s0022149x11000277 ◽

2011 ◽

Vol 86 (2) ◽

pp. 215-221 ◽

Cited By ~ 16

Author(s):

J.L. Ross ◽

E.S. Ivanova ◽

W.F. Sirgel ◽

A.P. Malan ◽

M.J. Wilson

Keyword(s):

South Africa ◽

18S Rrna ◽

Parasitic Nematode ◽

Gene Sequencing ◽

18S Rrna Gene ◽

Rrna Gene ◽

Western Cape ◽

Western Cape Province ◽

Rrna Gene Sequencing ◽

Three New Species

AbstractA survey of nematodes associated with native and introduced species of terrestrial slugs was conducted in the Western Cape Province of South Africa, in order to gather new data regarding diversity and distribution. A total of 521 terrestrial slugs were collected from 35 localities throughout the Western Cape. All slugs were dissected and examined for the presence of internal nematodes. Extracted nematodes were identified using a combination of molecular (18S rRNA gene sequencing) and morphological techniques. Nematodes were found parasitizing slugs at 14 of the 35 sites examined, amounting to 40% of sample sites. Of all slugs, 6% were infected with nematodes. A total of seven species of nematode were identified in the province, includingAgfa flexilis,Angiostomasp.,Phasmarhabditissp. SA1,Phasmarhabditissp. SA2,Caenorhabditis elegans,Panagrolaimussp. andRhabditissp. Of these species, four were thought to be parasitic to slugs (A. flexilis, Angiostomasp.,Phasmarhabditissp. SA1 andPhasmarhabditissp. SA2), as opposed to forming necromenic or phoretic associations. Three new species of slug-parasitic nematode were identified during this study (Angiostomasp.,Phasmarhabditissp. SA1 andPhasmarhabditissp. SA2).

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