scholarly journals Occurrence of different strains of Babesia canis in dogs in eastern Poland

2016 ◽  
Vol 60 (4) ◽  
pp. 423-427 ◽  
Author(s):  
Paweł Łyp ◽  
Michał Bartnicki ◽  
Marta Staniec ◽  
Stanisław Winiarczyk ◽  
Łukasz Adaszek
Keyword(s):  
Genetic Structure ◽  
Genetic Analysis ◽  
Geographical Distribution ◽  
18S Rrna ◽  
Clinical Course ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Babesia Canis ◽  
Pcr Products ◽  
Different Strains

Abstract Introduction: The aim of this study was to carry out a genetic analysis of Babesia canis isolates detected in dogs in eastern Poland and to study the correlation of the protozoa variant with a specific geographical region. Material and Methods: PCR was used to identify strains of B. canis from naturally infected animals (240 dogs from four provinces: Mazowieckie, Lublin, Podlasie, and Podkarpacie) by amplifying and sequencing a fragment of the 18S rRNA gene. Results: Sequencing the PCR products led to the identification of four variants of B. canis. Two previously described protozoa variants (18S rRNA-A and 18S rRNA-B) were observed in all provinces. Additionally, in the Mazowieckie and Lublin provinces a B. canis variant which contributed to the development of acute or atypical babesiosis was observed. The fourth variant of B. canis was detected only in dogs from the Lublin province, and the course of the disease was subclinical in all dogs infected with this variant. Conclusion: These results indicate the appearance of a new fourth B. canis genotype in Poland and confirm that it is still necessary to study the relationships between the genetic structure of protozoa, geographical distribution of the parasites, and clinical course of the disease.

Identification of new 18S rRNA strains of Babesia canis isolated from dogs with subclinical babesiosis

2015 ◽  
Vol 18 (3) ◽  
pp. 573-577 ◽  
Author(s):  
P. Łyp ◽  
Ł. Adaszek ◽  
B. Furmaga ◽  
S. Winiarczyk
Keyword(s):  
Ribosomal Rna ◽  
18S Rrna ◽  
Venous Blood ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Babesia Canis ◽  
Gene Fragment ◽  
Nucleotide Substitutions ◽  
New Variant ◽  
Pcr Products

Abstract In this study, we used PCR to detect and characterize B. canis from naturally infected dogs in Poland with subclinical babesiosis by amplifying and sequencing a portion of the 18S ribosomal RNA (rRNA) gene. Venous blood samples were collected from ten dogs with subclinical babesiosis. A 559-bp fragment of the B. canis 18S rRNA gene was amplified by PCR. Sequencing of the PCR products led to the identification of a new variant of Babesia canis, differing from the previously detected protozoa genotypes (18S rRNA-A and 18S rRNA-B) with nucleotide substitutions in positions 150 and 151 of the tested gene fragment. The results indicate the emergence within the Polish territory of a new, previously unencountered Babesia canis genotype responsible for the development of subclinical babesiosis.

scholarly journals Emergence of Babesia Conradae Infection in Coyote-hunting Greyhounds in Oklahoma, USA

2021 ◽  
Author(s):  
Erin M Stayton ◽  
Megan Lineberry ◽  
Jennifer Thomas ◽  
Tina Bass ◽  
Kelly Allen ◽  
...  
Keyword(s):  
18S Rrna ◽  
Clinical Signs ◽  
18S Rrna Gene ◽  
Post Treatment ◽  
Rrna Gene ◽  
Babesia Gibsoni ◽  
Wide Range ◽  
Pcr Products ◽  
Life Threatening ◽  
Tick Vectors

Abstract Background: Babesia species are intraerythrocytic Apicomplexan parasites that infect a wide range of vertebrate hosts. These pathogens are typically transmitted either by tick vectors or by direct blood-to-blood contact, and may cause life-threatening clinical disease such as thrombocytopenia, hemolytic anemia, and acute renal failure in canine hosts. While Babesia vogeli and Babesia gibsoni infections have both been reported in Oklahoma, reports of B. conradae infections have been limited to California. Methods: Whole blood samples were collected in EDTA tubes from all dogs in four separate kennels in Oklahoma. DNA was extracted from each blood sample and a nested PCR was performed using general Apicomplexan primers for the partial 18S rRNA gene. PCR products were electrophoresed in agarose matrix and appropriately sized amplicons were sequenced. Sequences were compared to reference 18S rRNA sequences available in GenBank, and samples with >98% homology to B. conradae (GenBank MK256976) were considered positive. B. conradae positive dogs were then treated with atovaquone (13.5 mg/kg TID) and azithromycin (10 mg/kg SID) for 10 days and retested at 30 and 60 days post treatment by PCR. Results: Fifteen of 40 dogs tested positive for B. conradae with 98–100% sequence homology to B. conradae from California. All positive cases were coyote-hunting Greyhounds. Treatment of clinically ill dogs with atovaquone and azithromycin resulted in complete clinical recovery in clinically ill dogs and all treated dogs had negative follow-up PCR at 30 and 60 days post treatment. Conclusions: Collectively, this study (i) documents the occurrence of B. conradae in Oklahoma, (ii) highlights this pathogen as a differential to be considered when clinical signs are present, and (iii) supports the use of atovaquone and azithromycin as effective treatment in these cases.

scholarly journals Phylogenetic information of freshwater copepod (Diaptomus sicilis) with special reference to 18S rRNA

2016 ◽  
Vol 4 (1) ◽  
pp. 25 ◽  
Author(s):  
Gomathi Jeyam Mookkaiah ◽  
Ramanibai Ravichandran
Keyword(s):  
Tamil Nadu ◽  
18S Rrna ◽  
Molecular Data ◽  
Small Subunit ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Calanoid Copepods ◽  
Universal Primer ◽  
Pcr Products ◽  
The Individual

<p>In the present investigation to isolate freshwater calanoid copepods (<em>Diaptomus sicilis</em>) was characterized and identify the organisms by 18S rRNA sequencing. Plankton samples containing <em>D. sicilis</em> were collected during January 2014 (Post-monsoon) from Madippakkam Lake (12°57'41"N80°11'27"E) Chennai, Tamil Nadu. Immediately after sampling, specimens for genetic analyses were fixed in 95% ethyl alcohol. The total DNA was extracted from the individual copepod <em>D. sicilis</em> using Qiagen Blood tissue kit. The nuclear small subunit 18S rRNA gene was amplified using the Universal primer LCO —1490 (5’-GGTCAACAAATCATAAAGATATTGG-3’) and HCO-2198 (5’-TAAACTTCAGGGTGACCAAAAAATCA-3’). PCR products were loaded onto a 1% TAE agarose gel. Sequences were carried out an automated sequencer. The nucleotide sequence of 1282 base pair region of 18S rRNA was determined for D. sicilis. The similarity of sequences of <em>D. sicilis</em> was retrieved by BLASTn pro­gram and maximum identity and E-value was 76% and 0.00, respectively. The PCR products of <em>D. sicilis</em> individuals showed 80% similarity with the partial nuclear small subunit 18S rRNA gene region of other calanoid copepods. Based on molecular data the freshwater Calanoid copepods showed different algorithms and similar types of topologies useful for designing molecular analyses using phylogeny tree construction.Present molecular stud­ies on the relationship of D. sicilis with other freshwater calanoid copepods indicate that this species is close to <em>D. castor</em> followed by <em>D. keniraensis</em><em>.</em></p>

scholarly journals Design of primer pairs for species-specific diagnosis of Leishmania (Leishmania) infantum chagasi using PCR

2012 ◽  
Vol 21 (3) ◽  
pp. 304-307 ◽  
Author(s):  
Osvaldo José da Silveira Neto ◽  
Sabrina Castilho Duarte ◽  
Hérika Xavier da Costa ◽  
Guido Fontgalland Coelho Linhares
Keyword(s):  
18S Rrna ◽  
18S Rrna Gene ◽  
Reference Sequence ◽  
Rrna Gene ◽  
Analytical Sensitivity ◽  
Specific Detection ◽  
Pcr Products ◽  
Specific Amplification ◽  
Pcr Assays ◽  
Species Specific

The objective of this study was to design and evaluate new primers for species-specific detection of L. infantum chagasi using PCR. Two combinations of primer pairs were established with the aim of obtaining specific amplification products from the L. infantum chagasi 18S rRNA gene. The combinations of the primer pairs and the respective sizes of the PCR products, based on the U422465 GenBank reference sequence of L. infantum chagasi, were: LCS1/LCS3 (259 bp) and LCS2/LCS3 (820 bp). It was concluded that the new PCR assays optimized using the primer pairs LCS1/LCS3 and LCS2/LCS3 were effective for specific detection of L. infantum chagasi, with analytical sensitivity to detect 1 pg/µL of DNA.

scholarly journals The diversity, distribution and host-parasite associations of trypanosomes in Western Australian wildlife

Parasitology ◽  
2009 ◽  
Vol 136 (11) ◽  
pp. 1269-1279 ◽  
Author(s):  
S. AVERIS ◽  
R. C. A. THOMPSON ◽  
A. J. LYMBERY ◽  
A. F. WAYNE ◽  
K. D. MORRIS ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Geographical Distribution ◽  
Host Species ◽  
18S Rrna ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Parasite Diversity ◽  
Host Parasite ◽  
Western Australian ◽  
Geographical Locations

SUMMARYLittle is known regarding the diversity, distribution or host-parasite associations of Trypanosoma spp. in Australian wildlife. Here we report on an investigation based on divergence of the 18S rRNA gene of trypanosomes isolated from a range of hosts and varied geographical locations. A total of 371 individuals representing 19 species of native animals from 14 different locations were screened. In total, 32 individuals from 9 different species tested positive for the parasite. Phylogenetic analysis revealed considerable parasite diversity with no clear geographical distribution and no evidence of host specificity. In general, it appears that Australian Trypanosoma spp. are widespread, with several genotypes appearing in multiple host species and in varied locations including both mainland areas and offshore islands. Some host species were found to be susceptible to multiple genotypes, but no individuals were infected with more than a single isolate.

Molecular Characterization and Assessment of Hematobiochemical and Oxidative Indices in Dog Naturally Infected with Babesia canis vogeli

2021 ◽  
Author(s):  
Prabhakar Shil ◽  
Jayesh B. Solanki ◽  
Niranjan Kumar ◽  
Dharmesh C. Patel ◽  
Nabanita Thakuria
Keyword(s):  
Plasma Cortisol ◽  
18S Rrna ◽  
Gene Sequence ◽  
Molecular Detection ◽  
18S Rrna Gene ◽  
Mean Value ◽  
Serum Glucose ◽  
Rrna Gene ◽  
Babesia Canis ◽  
The Mean

Background: The study was aimed at molecular detection and assessment of important biomarkers in the natural cases of canine babesiosis. Methods: Blood samples of 239 dogs were examined in PCR by targeting 18S rRNA gene. Hematobiochemical, oxidant-antioxidant and plasma cortisol parameters were estimated in the dogs on the day of presentation. Result: The 18S rRNA gene sequence showed 100% homology with Babesia canis vogeli and phylogram formed a tight cluster of B. canis vogeli originated from India/other countries. Higher prevalence rate (P less than 0.05) was noted in the PCR (7.95%) than the cytological technique (3.76%). Hemogram of infected dogs showed decrease (P less than 0.05) in the mean value of hemoglobin, RBC, WBC, HCT, whereas an increase in MCHC, lymphocytes, eosinophils, monocytes and thrombocytes. The ALT (49.29±1.53 U/L), AST (48.33±2.93 U/L), total protein (10.56±0.60 g/dL), creatinine (1.41±0.10 mg/dL) and urea (19.32±0.97 mg/dL) showed significant (P less than 0.005) increase, whereas decrease in the levels of serum glucose (82.76±2.78 mg/dL) in the infected dogs. Activity of MDA and SOD was significantly (P less than 0.01) increased (7.50±7.08 nmole/µL blood) and decreased (0.015±3.91 nmole/µL blood) in the diseased dogs, respectively. Plasma cortisol concentration was 11.10±7.84 nmol/L and 2.77±5.78 nmol/L (P less than 0.01) in the infected and uninfected dogs, respectively.

scholarly journals Phylogenetic Analysis of the Hypervariable Region of the 18S rRNA Gene of Cryptosporidium Oocysts in Feces of Canada Geese (Branta canadensis): Evidence for Five Novel Genotypes

2004 ◽  
Vol 70 (1) ◽  
pp. 452-458 ◽  
Author(s):  
Kristen L. Jellison ◽  
Daniel L. Distel ◽  
Harold F. Hemond ◽  
David B. Schauer
Keyword(s):  
Genetic Diversity ◽  
Phylogenetic Analysis ◽  
18S Rrna ◽  
Hypervariable Region ◽  
Pcr Amplification ◽  
The United States ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Canada Geese ◽  
Pcr Products

ABSTRACT To assess genetic diversity in Cryptosporidium oocysts from Canada geese, 161 fecal samples from Canada geese in the United States were analyzed. Eleven (6.8%) were positive for Cryptosporidium spp. following nested PCR amplification of the hypervariable region of the 18S rRNA gene. Nine PCR products from geese were cloned and sequenced, and all nine diverged from previously reported Cryptosporidium 18S rRNA gene sequences. Five sequences were very similar or identical to each other but genetically distinct from that of Cryptosporidium baileyi; two were most closely related to, but genetically distinct from, the first five; and two were distinct from any other sequence analyzed. One additional sequence in the hypervariable region of the 18S rRNA gene isolated from a cormorant was identical to that of C. baileyi. Phylogenetic analysis provided evidence for new genotypes of Cryptosporidium species in Canada geese. Results of this study suggest that the taxonomy of Cryptosporidium species in geese is complex and that a more complete understanding of genetic diversity among these parasites will facilitate our understanding of oocyst sources and species in the environment.

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.

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