scholarly journals Molecular Detection of Tick-Borne Agents in Cats from Southeastern and Northern Brazil

Pathogens ◽  
2022 ◽  
Vol 11 (1) ◽  
pp. 106
Author(s):  
Marcos Rogério André ◽  
Ana Cláudia Calchi ◽  
Maria Eduarda Chiaradia Furquim ◽  
Isabela de Andrade ◽  
Paulo Vitor Cadina Arantes ◽  
...  
Keyword(s):  
18S Rrna ◽  
Clinical Signs ◽  
18S Rrna Gene ◽  
Molecular Techniques ◽  
Rrna Gene ◽  
Molecular Identity ◽  
Northern Brazil ◽  
Rrna Gene Sequencing ◽  
First Time ◽  
Theileria Spp

Even though the epidemiology of tick-borne agents (TBA) in dogs has been extensively investigated around the world, the occurrence, vectors involved, and molecular identity of these agents in cats remains elusive in many regions. Among TBA, Ehrlichia, Anaplasma, Babesia, Cytauxzoon, and Hepatozoon are responsible for diseases with non-specific clinical signs in cats, making essential the use of molecular techniques for accurate diagnosis and proper treatment. The present work aimed to investigate the occurrence and molecular identity of tick-borne agents (Ehrlichia, Anaplasma, Babesia/Theileria, Cytauxzoon, and Hepatozoon) in cats from southeastern (states of São Paulo (SP) and Minas Gerais (MG)) and northern (state of Rondônia (RO)) Brazil. For this purpose, 390 blood samples were collected from domiciled cats in MG (n = 155), SP (n = 151), and RO(n = 84) states, submitted to DNA extraction and PCR assays for Ehrlichia spp. (dsb gene), Anaplasma spp. (rrs gene), piroplasmids (18S rRNA gene), and Hepatozoon spp. (18S rRNA gene), sequencing, and phylogenetic inferences. The overall positivity for Anaplasma spp., Ehrlichia spp., Babesia/Theileria spp., Cytauxzoon spp., and Hepatozoon spp. were 7.4% (12.3% (MG) and 6.6% (SP)), 2% (4.5% (MG) and 0.6% (SP)), 0.7% (0.6% (MG), 0.6% (SP) and 1.2% (RO)), 27.2% (41.9% (MG), 24.5% (SP) and 4.8% (RO), and 0%, respectively. The phylogenetic analysis grouped the obtained sequences with ‘Candidatus Anaplasma amazonensis’, A. platys, B. vogeli, and Cytauxzoon sp. previously detected in wild felids from Brazil. qPCR specific for E. canis based on the dsb gene confirmed the molecular identity of the detected ehrlichial agent. The present study expanded the list and geographical distribution of hemoparasites in cats. ‘Candidatus Anaplasma amazonensis’, recently detected in sloths from northern Brazil, was described for the first time in cats. This is the first report of piroplasmids infecting cats in northern Brazil. Coinfection by Cytauxzoon and other TBA (Ehrlichia, Anaplasma, and B. vogeli) reported in the present study raises the need for veterinary practitioners’ awareness of cats parasitized by multiple TBA.

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

2019 ◽  
Vol 131 ◽  
pp. 01043 ◽  
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.

Association between diet and rumen microbiota in wild roe deer

2019 ◽  
Vol 366 (6) ◽  
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.

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

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.

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

2011 ◽  
Vol 86 (2) ◽  
pp. 215-221 ◽  
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).

18S rRNA gene sequencing identifies a novel species of Henneguya parasitizing the gills of the channel catfish (Ictaluridae)

2014 ◽  
Vol 113 (12) ◽  
pp. 4651-4658 ◽  
Author(s):  
Thomas G. Rosser ◽  
Matt J. Griffin ◽  
Sylvie M. A. Quiniou ◽  
Lester H. Khoo ◽  
Linda M. Pote
Keyword(s):  
Channel Catfish ◽  
18S Rrna ◽  
Novel Species ◽  
Gene Sequencing ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Rrna Gene Sequencing

scholarly journals Babesia microti in Rodents from Different Habitats of Lithuania

Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1707
Author(s):  
Dalytė Mardosaitė-Busaitienė ◽  
Jana Radzijevskaja ◽  
Linas Balčiauskas ◽  
Algimantas Paulauskas
Keyword(s):  
18S Rrna ◽  
18S Rrna Gene ◽  
Reservoir Host ◽  
Clethrionomys Glareolus ◽  
Babesia Microti ◽  
Rrna Gene ◽  
Apodemus Flavicollis ◽  
Study Sites ◽  
First Time ◽  
Partial 18S

Babesia microti (Aconoidasida: Piroplasmida) (Franca, 1910) is an emerging tick-borne parasite with rodents serving as the considered reservoir host. However, the distribution of B. microti in Europe is insufficiently characterized. Based on the sample of 1180 rodents from 19 study sites in Lithuania, the objectives of this study were: (1) to investigate the presence of Babesia parasites in eight species of rodents, (2) to determine the prevalence of Babesia parasites in rodents from different habitats, and (3) to characterize the detected Babesia strains using partial sequencing of the 18S rRNR gene. Babesia DNA was detected in 2.8% rodents. The highest prevalence of Babesia was found in Microtus oeconomus (14.5%) and Microtus agrestis (7.1%) followed by Clethrionomys glareolus (2.3%), Apodemus flavicollis (2.2%) and Micromys minutus (1.3%). In M. minutus, Babesia was identified for the first time. The prevalence of Babesia-infected rodents was higher in the meadow (5.67%) than in the ecotone (1.69%) and forest (0.31%) habitats. The sequence analysis of the partial 18S rRNA gene reveals that Babesia isolates derived from rodents were 99–100% identical to human pathogenic B. microti ‘Jena/Germany’ strain.

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