Molecular detection and genetic diversity of Anaplasma marginale based on the major surface protein genes in Thailand

Junsiri, Witchuta; Watthanadirek, Amaya; Poolsawat, Napassorn; Kaewmongkol, Sarawan; Jittapalapong, Sathaporn; Chawengkirttikul, Runglawan; Anuracpreeda, Panat

doi:10.1016/j.actatropica.2020.105338

Cited by 24 publications

(15 citation statements)

References 32 publications

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“…DNA samples were re-confirmed by Anaplasmataceae -specific primers EHR16SR (5′-TAG-CAC-TCA-TCG-TTT-ACA-GC-3′) and EHR16SD (5′-GGT-ACC-YAC-AGA-AGA-AGT-CC-3′) [ 19 ]. Other species-specific primers targeting the major surface protein 2 gene of A. marginale , MSP2-F (5′-CAC-CAT-GAG-TGC-TGT-AAG-TAA-TAG-GAA-GC-3′) and MSP2-R (5′-CTA-GAA-GGC-AAA-CCT-AAC-ACC-CAA-CTC-3′) were used to identify the A. marginale in rickettsial organism-positive samples in this study [ 39 ]. All A. platys -like species detected in buffalo were also verified using the species-specific primers PLATYS-F (5′-AAG-TCG-AAC-GGA-TTT-TTG-TC-3′) and PLATYS-R (5′-CTT-TAA-CTT-ACC-GAA-CC-3′) [ 40 ].…”

Section: Methodsmentioning

confidence: 99%

Molecular detection and genetic characterization of Anaplasma marginale and Anaplasma platys-like (Rickettsiales: Anaplasmataceae) in water buffalo from eight provinces of Thailand

2020

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Background Anaplasmosis, an animal disease caused by rickettsial bacteria in the genus Anaplasma, is of considerable economic importance in livestock animals in many countries worldwide. The objectives of this study were to determine the identity, prevalence, and geographic distribution of Ehrlichia and Anaplasma in naturally infected water buffalo in Thailand using PCR amplification and sequencing of the 16S ribosomal RNA and heat shock protein groEL genes. A total of 456 buffalo blood samples from Thailand were investigated. Species identification and genetic differentiation of intra-population and inter-population with the global isolates were conducted based on nucleotide sequences. Interplay between the infection and host factors was also assessed. Results Overall, 41% of water buffalo were found to be infected with rickettsial organisms in the family Anaplasmataceae, but Ehrlichia spp., Neorickettsia spp., and Wolbachia spp. were not found in any of the sequenced samples in this study. Female buffalo were more frequently infected with bacteria in the family Anaplasmataceae than males [71 out of 176 females (40.3%) versus 11 out of 47 males (23.4%)]. The Odds Ratio value indicated that the risk of infection for female buffalo was 2.2-fold higher than that for males (p < 0.05). We detected three haplotypes of A. marginale 16S rRNA gene and they were placed in a clade that was closely related to the A. marginale in buffalo in China; and cattle in Thailand, Uganda, and China. Homology searching of groEL sequences against the GenBank™ database using the BLASTn algorithm revealed that the obtained sequences had a high percentage similarity (98.36–99.62%) to A. platys sequences. The groEL sequences of three A. platys-like isolates were clustered in the same clade as the A. platys from the tick Rhipicephalus microplus in China. Conclusions Our data showed that the apparently healthy buffalo were naturally infected by bacteria in the family Anaplasmataceae at a relatively high prevalence. We also report the finding of A. platys-like infections in water buffalo in Thailand for the first time. Water buffalo serving as the reservoir host of anaplasmosis is of concern for managing the disease control and prevention in ruminants.

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Section: Methodsmentioning

confidence: 99%

Molecular detection and genetic characterization of Anaplasma marginale and Anaplasma platys-like (Rickettsiales: Anaplasmataceae) in water buffalo from eight provinces of Thailand

2020

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“…Genomic DNA of canine TBPs (H. canis, A. platys and E. canis) was extracted from 250 μL of dogs' blood samples using a Tissue DNA Extraction Kit (OMEGA, bio-tex, USA) following the method of Watthanadirek et al (2019) and Junsiri et al (2020) with some modifications. Briefly, 250 μL of each blood sample was transferred to sterile microcentrifuge tubes.…”

Section: Dna Extractionmentioning

confidence: 99%

Molecular discrimination and genetic diversity of three common tick-borne pathogens in dogs in Thailand

Poolsawat

Tazawa²,

Junsiri

et al. 2021

Parasitology

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There was little information regarding the occurrence of canine vector-borne disease (CVBDs) in shelter dogs in Thailand. This work is the first report regarding a molecular method used to determine the occurrence and genetic diversity of three canine tick-borne pathogens (TBPs) (Hepatozoon canis, Anaplasma platys and Ehrlichia canis) in blood samples from 275 shelter dogs in the north and central areas of Thailand. The PCR results based on the 18S rRNA and 16S rRNA genes showed that 71 (25.82%) dogs were positive for at least a TBP. The overall occurrence rates of H. canis, A. platys and E. canis infections were 1.81, 16.36 and 7.64%, respectively. For the phylogenetic analysis, A. platys 16S rRNA gene was genetically diverse, while H. canis 18S rRNA and E. canis 16S rRNA genes were conserved. The haplotype diversity exhibited 12 and 2 haplotypes as well as 78 and 178 polymorphic sites of A. platys and E. canis 16S rRNA genes, respectively. Our findings could be used to improve the understanding of phylogeny and genetic diversity of TBP rRNA genes and used to ameliorate the diagnosis and control programmes for the diseases in Thailand.

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“…Genomic DNA of L. sabrazesi was extracted from all collected blood samples of chickens using Tissue DNA Extraction Kit (OMEGA, bio-tek, USA) following the protocol of Watthanadirek et al 32 and Junsiri et al 33 with some modifications. Briefly, 20 µl of blood samples were mixed with 25 µl of OB Protease solution and 250 µl of BL buffer and incubated at 70 °C for 1 min.…”

Section: Methodsmentioning

confidence: 99%

“…The positive clones were selected and grown in LB medium containing ampicillin for overnight. Finally, the recombinant plasmids (pET100- cytb ) were extracted from the competent cells using AXYPREP Plasmid Miniprep Kit (AXYGEN BIOSCIENCE, USA) following the manufacturer’s instructions and analyzed for correctly sized inserts by agarose gel electrophoresis 32 , 33 .…”

Section: Methodsmentioning

confidence: 99%

Molecular detection and genetic diversity of Leucocytozoon sabrazesi in chickens in Thailand

Chawengkirttikul

Junsiri

Watthanadirek

et al. 2021

Sci Rep

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Leucocytozoon sabrazesi is the intracellular protozoa of leucocytozoonosis, which is transmitted by the insect vectors and affects chickens in most subtropical and tropical regions of the globe, except South America, and causing enormous economic losses due to decreasing meat yield and egg production. In this study, L. sabrazesi gametocytes have been observed in the blood smears, and molecular methods have been used to analyse the occurrence and genetic diversity of L. sabrazesi in blood samples from 313 chickens raised in northern, western and southern parts of Thailand. The nested polymerase chain reaction (nested PCR) assay based on the cytb gene revealed that 80.51% (252/313) chickens were positive of L. sabrazesi. The phylogenetic analysis indicated that L. sabrazesi cytb gene is conserved in Thailand, showed 2 clades and 2 subclades with similarity ranged from 89.5 to 100%. The diversity analysis showed 13 and 18 haplotypes of the sequences from Thailand and from other countries, respectively. The entropy analyses of nucleic acid sequences showed 26 high entropy peaks with values ranging from 0.24493 to 1.21056, while those of amino acid sequences exhibited 5 high entropy peaks with values ranging from 0.39267 to 0.97012. The results; therefore, indicate a high molecular occurrence of L. sabrazesi in chicken blood samples with the associated factors that is statistically significant (p < 0.05). Hence, our results could be used to improve the immunodiagnostic methods and to find appropriate preventive control strategies or vaccination programs against leucocytozoonosis in order to mitigate or eliminate the harmful impact of this infection on chicken industry.

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Molecular detection and genetic diversity of Anaplasma marginale based on the major surface protein genes in Thailand

Cited by 24 publications

References 32 publications

Molecular detection and genetic characterization of Anaplasma marginale and Anaplasma platys-like (Rickettsiales: Anaplasmataceae) in water buffalo from eight provinces of Thailand

Molecular detection and genetic characterization of Anaplasma marginale and Anaplasma platys-like (Rickettsiales: Anaplasmataceae) in water buffalo from eight provinces of Thailand

Molecular discrimination and genetic diversity of three common tick-borne pathogens in dogs in Thailand

Molecular detection and genetic diversity of Leucocytozoon sabrazesi in chickens in Thailand

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