Recombinant expression and characterization of major surface protein 4 from Anaplasma marginale

Watthanadirek, Amaya; Chawengkirttikul, Runglawan; Poolsawat, Napassorn; Junsiri, Witchuta; Boonmekam, Dusit; Reamtong, Onrapak; Anuracpreeda, Panat

doi:10.1016/j.actatropica.2019.105047

Cited by 17 publications

(8 citation statements)

References 35 publications

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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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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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“…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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“…The genomic DNA of L. sabrazesi in blood samples was extracted by using an E.Z.N.A. ® Tissue DNA Kit (OMEGA Bio-Tek, Norcross, GA, USA) following the protocol of Watthanadirek et al [ 42 , 43 ] and Junsiri [ 22 ] with some modifications. Briefly, 250 μL of blood samples were mixed thoroughly with 25 μL of proteinase K solution, and incubated at 70 °C for 10 min.…”

Section: Methodsmentioning

confidence: 99%

Molecular genetic diversity and bioinformatic analysis of Leucocytozoon sabrazesi based on the mitochondrial genes cytb, coxI and coxIII and co-infection of Plasmodium spp.

et al. 2022

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Leucocytozoon sabrazesi is an intracellular haemoprotozoan parasite responsible for leucocytozoonosis, which is transmitted by insect vectors and affects chickens in tropical and subtropical areas in many countries. It causes huge economic losses due to decreased meat and egg production. In the present study, we used nested PCR to determine the genetic diversity of L. sabrazesi based on the cytb, coxI, coxIII and concatenated genes in chickens in Thailand. In addition, we found co-infections between L. sabrazesi and Plasmodium spp. (P. gallinaceum or P. juxtanucleare) in chickens that were not identified by microscopic examination of blood smears. The phylogenetic analysis indicated that L. sabrazesi cytb and coxIII genes were conserved with similarity ranging from 99.9 to 100% and 98 to 100%, respectively whereas the coxI gene was diverse, with similarities ranging from 97 to 100%. These findings ascertained the nucleotide analysis of the cytb, coxI, coxIII and concatenated sequences in which 4, 8, 10 and 9 haplotypes were found, respectively. In addition, it was found that the large number of synonymous substitutions and conservative amino acid replacements in these mitochondrial genes occurred by non-synonymous substitution. The evolutionary analysis of the Ka/Ks ratio supported purifying selection and the negative values of both Fu’s Fs and Tajima’s D indicate selective sweep especially for the coxI gene. The entropy and Simplot analysis showed that the genetic variation in populations of Plasmodium spp. was higher than in Leucocytozoon. Hence, the nucleotide sequences of three mitochondrial genes could reflect the evolutionary analysis and geographic distribution of this protozoan population that switches hosts during its life cycle.

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Recombinant expression and characterization of major surface protein 4 from Anaplasma marginale

Cited by 17 publications

References 35 publications

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

Molecular genetic diversity and bioinformatic analysis of Leucocytozoon sabrazesi based on the mitochondrial genes cytb, coxI and coxIII and co-infection of Plasmodium spp.

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