First Report Regarding the Simultaneous Molecular Detection of Anaplasma marginale and Theileria annulata in Equine Blood Samples Collected from Southern Punjab in Pakistan.
Abstract: The present study was designed to check the molecular detection of Anaplasma marginale and Theileria annulata in blood samples of horses and donkeys collected from Dera Ghazi Khan District in Punjab and to document their phylogenetic origin and their association with studied epidemiological factors (sex and age) and complete blood count parameters, if any. Results: A total of 195 blood samples were collected from apparently healthy horses (N = 141) and donkeys (N = 54). A. marginale DNA was detected by PCR in 4.9% (7/141) horse and in 9.2% (5/54) of donkey blood samples. Prevalence of T. annulata was 5.6% (8/141) and 11.1% (6/54) in horse and donkey samples, respectively. While 1.4% (N = 2) horses and 3.7% (N = 2) donkeys were found co-infected with both parasites. Representative amplicon for both parasites was confirmed by DNA sequenced and partial DNA sequence of the major surface protein-1b encoding gene of A. marginale and cytochrome b gene from T. annulata were submitted to the GenBank database under the accession number MK792344-MK792348. Epidemiological data analysis revealed that female horses were more prone to A. marginale (P = 0.02) while female donkeys were more susceptible to A. marginale (P < 0.001) and T. annulata (P < 0.001) infection. It was observed that horse and donkey infected either with Anaplasma marginale or Theileria annulata had significantly disturbed red and white blood cell counts and their associated parameters. Conclusions: This is a first ever study regarding molecular detection of A. marginale and T. annulata in equine blood samples from Pakistan. We recommend that this multiplex PCR protocol should be used for the detection of Anaplasma marginale and Theileria annulata in livestock for their proper diagnosis and treatment.
Publication Date: 2019-11-13 PubMed ID: 31721059DOI: 10.2478/s11686-019-00141-wGoogle Scholar: Lookup
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- Journal Article
Summary
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The research article reports the first molecular detection of Anaplasma marginale and Theileria annulata infections in horses and donkeys in Southern Punjab, Pakistan. The study also explores the relationship between the infections and specific epidemiological factors and blood count parameters.
Objective and Methodology of the Study
- The primary objective of this research was to conduct molecular detection of two parasites, Anaplasma marginale and Theileria annulata, in the blood of horses and donkeys from the Dera Ghazi Khan District in Punjab.
- The researchers aimed to understand the phylogenetic origin of these parasites and their correlation with certain epidemiological factors such as sex and age. They also examined whether these infections influenced complete blood count parameters in the host.
- In total, 195 blood samples were collected from seemingly healthy horses and donkeys (141 horses and 54 donkeys).
- The presence of the parasites was determined using PCR (Polymerase Chain Reaction), a common method to detect DNA in samples.
Results and Findings
- DNA evidence of A. marginale was found in 4.9% of horse samples (7 out of 141) and 9.2% of donkey samples (5 out of 54).
- T. annulata was present in 5.6% of horse samples (8 out of 141) and 11.1% of donkey samples (6 out of 54).
- Co-infection, where both parasites were present, was identified in 1.4% of horses (2 out of 141) and 3.7% of donkeys (2 out of 54).
- The extracted DNA from both parasites was sequenced and submitted to the GenBank database for further studies.
- The study highlighted a gender disparity in infection susceptibility, with female horses and donkeys found to be more prone to infection by these parasites. No specific significance was given as to why this gender bias occurred.
- Evidence suggested that horses and donkeys infected with either parasite showed significant anomalies with their red and white blood cell counts and related parameters.
Conclusion and Recommendation
- This research is the first to report molecular detection of A. marginale and T. annulata in equine blood samples from Pakistan.
- The authors recommend the use of this multiplex PCR protocol for accurate diagnosis and appropriate treatment of A. marginale and T. annulata infections in livestock.
Cite This Article
APA
Asif M, Parveen A, Ashraf S, Hussain M, Aktas M, Ozubek S, Shaikh RS, Iqbal F.
(2019).
First Report Regarding the Simultaneous Molecular Detection of Anaplasma marginale and Theileria annulata in Equine Blood Samples Collected from Southern Punjab in Pakistan.
Acta Parasitol, 65(1), 259-263.
https://doi.org/10.2478/s11686-019-00141-w Publication
Researcher Affiliations
- Institute of Molecular Biology and Biotechnology, Bahauddin Zakariya University, Multan, 60800, Pakistan.
- Zoology Division, Institute of Pure and Applied Biology, Bahauddin Zakariya University, Multan, 60800, Pakistan.
- Zoology Division, Institute of Pure and Applied Biology, Bahauddin Zakariya University, Multan, 60800, Pakistan.
- Institute of Molecular Biology and Biotechnology, Bahauddin Zakariya University, Multan, 60800, Pakistan.
- Department of Parasitology, Veterinary Faculty, Fırat University, 23119, Elazig, Turkey.
- Department of Parasitology, Veterinary Faculty, Fırat University, 23119, Elazig, Turkey.
- Institute of Molecular Biology and Biotechnology, Bahauddin Zakariya University, Multan, 60800, Pakistan. rehansadiq80@bzu.edu.pk.
- Zoology Division, Institute of Pure and Applied Biology, Bahauddin Zakariya University, Multan, 60800, Pakistan. furhan.iqbal@bzu.edu.pk.
MeSH Terms
- Anaplasma marginale / isolation & purification
- Anaplasmosis / blood
- Anaplasmosis / diagnosis
- Anaplasmosis / epidemiology
- Animals
- DNA, Bacterial / genetics
- DNA, Protozoan / genetics
- Equidae / parasitology
- Female
- Horse Diseases / blood
- Horse Diseases / diagnosis
- Horse Diseases / epidemiology
- Horses / parasitology
- Male
- Multiplex Polymerase Chain Reaction
- Pakistan / epidemiology
- Phylogeny
- Prevalence
- Protozoan Proteins / genetics
- RNA, Ribosomal, 18S / genetics
- Theileria annulata / isolation & purification
- Theileriasis / blood
- Theileriasis / diagnosis
- Theileriasis / epidemiology
Grant Funding
- Ph D Fellowship / Higher Education Commision, Pakistan
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Citations
This article has been cited 4 times.- de la Fournière S, Guillemi EC, Paoletta MS, Pérez A, Obregón D, Cabezas-Cruz A, Sarmiento NF, Farber MD. Transovarial Transmission of Anaplasma marginale in Rhipicephalus (Boophilus) microplus Ticks Results in a Bottleneck for Strain Diversity.. Pathogens 2023 Aug 2;12(8).
- Atif FA, Abbas RZ, Mehnaz S, Qamar MF, Hussain K, Nazir MU, Zaman MA, Khan AU, Said MB. First report on molecular surveillance based on duplex detection of Anaplasma marginale and Theileria annulata in dairy cattle from Punjab, Pakistan.. Trop Anim Health Prod 2022 Apr 1;54(2):155.
- Parveen A, Alkhaibari AM, Asif M, Almohammed HI, Naqvi Z, Khan A, Aktas M, Ozubek S, Farooq M, Iqbal F. Molecular Epidemiology of Theileria annulata in Cattle from Two Districts in Punjab (Pakistan).. Animals (Basel) 2021 Dec 2;11(12).
- Parveen A, Ashraf S, Aktas M, Ozubek S, Iqbal F. Molecular epidemiology of Theileria annulata infection of cattle in Layyah District, Pakistan.. Exp Appl Acarol 2021 Mar;83(3):461-473.
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