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Molecular detection of Theileria annulata infection in cattle by conventional PCR and quantitative real time PCR in India

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Abstract

Theileria annulata (T. annulata) is a tick-borne apicomplexan parasite that affects bovine. It is endemic in many tropical and subtropics areas, including Odisha, India. The objective of this study is to identify T. annulata infection in the peripheral blood of cattle as a biological sample by conventional PCR (cPCR) and quantitative PCR (qPCR). The phylogenetic analysis was done using the T. annulata merozoite surface antigen (Tams 1) gene. Out of 552 samples of examined blood smears by microscopy, 454 (82.24%) animals were positive for Theileria species. Out of 454 samples, 96 samples were further examined by both cPCR and qPCR, 52 samples (54.16%) were found positive for T. annulata in both PCR methodologies. Phylogenetic analysis revealed that T. annulata Odisha isolate was closely related to T. annulata Uttarakhand, India isolate (KM061799) and Hyderabad, India isolate (MK034702) with Nucleotide sequence identity 95.36%, 95.25%, respectively. This is the first study to detect T. annulata by qPCR in Odisha and supported that both PCR techniques were equally effective for the detection of Tams 1 gene of T. annulata in cattle’s blood.

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References

  • Acharya AP, Panda SK, Prusty BK (2017) Diagnosis and confirmation of Theileria annulata infection in cattle in Odisha, India. J Entomol Zool Stud 5(4):1543–1546

    Google Scholar 

  • Ananda KJ, D’Souza P, Puttalakshmamma GC (2009) Prevalence of Haemoprotozoan diseases in crossbred cattle in Bangalore north. Vet World 2:5–16

    Article  Google Scholar 

  • Azimi S, Mahravani H, Jeirani F, Shoshtari A (2011) Appling real time RT-PCR for bluetongue virus detection in Iran. Archi Razi 66:75–80

    Google Scholar 

  • Bilgic HB, Karagenc T, Shiels B, Tait A, Eren H, Weir W (2010) Evaluation of cytochrome b as a sensitive target for PCR based detection of T. annulata carrier animals. Vet Parasitol 174:341–347

    Article  CAS  Google Scholar 

  • Bogema DR, Deutscher AT, Fell S, Collins D, Eamens GJ, Jenkins C (2015) Development and validation of a quantitative PCR assay using multiplexed hydrolysis probes for detection and quantification of Theileria orientalis isolates and differentiation of clinically relevant subtypes. J Clin Microbiol 53:941–950

    Article  CAS  Google Scholar 

  • Branco S, Orvalho J, Leitao A, Pereira I, Malta M, Mariano I, Carvalho T, Baptista R, Shiels BR, Peleteiro MC (2010) Fatal cases of Theileria annulata infection in calves in Portugal associated with neoplastic-like lymphoid cell proliferation. J Vet Sci 11:27–34

    Article  Google Scholar 

  • d’Oliveira C, Van Der Weide M, Habela P, Jacquiet P, Jongejan F (1995) Detection of Theileria annulata in blood samples of carrier cattle by PCR. J Clin Microbiol 13:2665–2669

    Article  Google Scholar 

  • Dandasena D, Bhandari V, Sreenivasamurthy GS, Murthy S, Roy S, Bhanot V, Singh S, Sharma P (2018) A Real-Time PCR based assay for determining parasite to host ratio and parasitaemia in the clinical samples of Bovine Theileriosis. Sci Rep 8(1):15441

    Article  Google Scholar 

  • Demessie Y, Derso S (2015) Tick Borne Hemoparasitic Diseases of Ruminants: A Review. Adv Bio Res 9:210–224

    CAS  Google Scholar 

  • Gebrekidan H, Gasser RB, Stevenson MA, Jabbar A (2018) Multiplexed tandem PCR (MT-PCR) assay using the major piroplasm surface protein gene for the diagnosis of Theileria orientalis infection in cattle. J Clin Microbiol 56:1661–1671

    Google Scholar 

  • Ghaemi P, Hoghooghi-Rad N, Shayan P, Eckert B (2012) Detection of Theileria orientalis in Iran by semi-nested PCR. Parasitol Res 110:527–531

    Article  Google Scholar 

  • Godara R, Sharma RL, Sharma CS (2009) Bovine Tropical Theileriosis in a neonate calf. Trop Anim Health Prod 42:551–553

    Article  Google Scholar 

  • Jain K (1993) Essentials of Veterinary Hematology. Blackwell Publishing, U.S.A

    Google Scholar 

  • Kearns A, Guiver M, James V, King J (2001) Development and evaluation of a real-time quantitative PCR for the detection of human cytomegalovirus. J Virol Methods 95:121–131

    Article  CAS  Google Scholar 

  • Kundave VR, Patel AK, Patel PV, Hasnani JJ, Joshi CG (2014) Qualitative and quantitative assessment of Theileria annulata in cattle and buffaloes by Polymerase Chain Reaction. Trop Biomed 31(4):728–735

    CAS  PubMed  Google Scholar 

  • Mohammad Al-Saeed AT, Omer LT, Abdo J, Habibi G, Salih DA, Seitzer U, Ahmed J (2010) Epidemiological studies on tropical theileriosis (Theileria annulata infection of cattle) in Kurdistan Region, Iraq. Parasitol Res 106(2):403–407

    Article  Google Scholar 

  • Nazifi S, Razavi SM, Esmailnejad Z, Gheisari H (2009) Study on acute phase proteins (haptoglobin, serum amyloid A, fibrinogen, and ceruloplasmin) changes and their diagnostic values in bovine tropical theileriosis. Parasitol Res 105:41–46

    Article  CAS  Google Scholar 

  • Niedbalski W, Ksy A, Procajo Z, Szweda W, Mikulska SE, Platt SA, Guo-Wen L, Zhi-Gang Z, Wen-Yan Y, Lian-Yu Y (2010) Rapid detection and quantification of foot-and-mouth disease virus by a real-time reverse transcription PCR. B Vet I Pulawy 54

  • Pereira MR, Rocha-Silva F, Graciele-Melo C, Lafuente CR, Magalhães T, Caligiorne RB (2014) Comparison between conventional and real-time PCR assays for diagnosis of visceral leishmaniasis. Biomed Res Int 63:93–100

    Google Scholar 

  • Perera PK, Gasser RB, Firestone SM, Smith L, Roeber F, Jabbar A (2015) A semiquantitative multiplexed tandem PCR for detection and differentiation of four Theileria orientalis genotypes in cattle. J Clin Microbiol 53:79–87

    Article  CAS  Google Scholar 

  • Ros-García A, Nicolás A, García-Pérez AL, Juste RA, Hurtado A (2012) Development and evaluation of a real-time PCR assay for the quantitative detection of Theileria annulata in cattle. Parasit Vectors 5:171

    Article  Google Scholar 

  • Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425

    CAS  PubMed  Google Scholar 

  • Santos M, Soares R, Costa P (2013) Revisiting the Tams1-encoding gene as a species-specific target for the molecular detection of Theileria annulata in bovine blood samples. Ticks Tick Borne Dis 4:72–77

    Article  Google Scholar 

  • Sengupta PP, Bansal GC, Ray D (1993) Ocular lesions in experimental theileriosis. J Vet Parasitol 7:127–129

    Google Scholar 

  • Sitotaw T, Regassa F, Zeru F, Kahsay AG (2014) Epidemiological significance of major hemoparasites of ruminants in and around Debre-Zeit, Central Ethiopia. J Parasitol Vector Bio 6:16–22

    Google Scholar 

  • Sivakumar T, Kothalawala H, Abeyratne SA, Vimalakumar SC, Meewewa AS, Hadirampela DT, Puvirajan T, Sukumar S, Kuleswarakumar K, Chandrasiri AD, Igarashi I, Yokoyama N (2012) A PCR-based survey of selected Babesia and Theileria parasites in cattle in Sri Lanka.Vet Parasitol 190(1–2):263–267

  • Sudan V, Sharma RL, Yadav R, Borah MK (2012) Turning sickness in a cross bred cow naturally infected with Theileria annulata. J Parasit Dis 36:226–229

    Article  Google Scholar 

  • Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA 6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30(12):2725–2729

    Article  CAS  Google Scholar 

  • Zaeemi M, Haddadzadeh H, Khazraiinia P, Kazemi B, Bandehpour M (2011) Identification of different Theileria species (Theileria lestoquardi, Theileria ovis, and Theileria annulata) in naturally infected sheep using nested PCR–RFLP. Parasitol Res 108:837–843

    Article  Google Scholar 

  • Zarlenga DS, Higgins J (2001) PCR as adiagnostic and quantitative technique in veterinary parasitology. Vet Parasitol 101:215–230

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Clinical Medicine, College of Veterinary Science and Animal Husbandry, Odisha University of Agriculture and Technology, Bhubaneshwar, Odisha, 751003, India

    Ahmed Magdy Selim, Manoranjan Das, Santosh Kumar Senapati & Geeta Rani Jena

  2. Internal Medicine, Infectious Diseases and Fish Diseases Department, Faculty of Veterinary Medicine, Mansoura University, 35516, Mansoura, Egypt

    Ahmed Magdy Selim

  3. Department of Animal Breeding and Genetics, College of Veterinary Science and Animal Husbandry, Odisha University of Agriculture and Technology, Bhubaneshwar, Odisha, 751003, India

    Chinmoy Mishra

  4. Department of Veterinary Surgery, College of Veterinary Science and Animal Husbandry, Odisha University of Agriculture and Technology, Bhubaneshwar, Odisha, 751003, India

    Indramani Nath

  5. Institute of Life Sciences, Bhubaneswar, Odisha, India

    Shantibhusan Senapati & Manisha Sethi

Authors
  1. Ahmed Magdy Selim
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  2. Manoranjan Das
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  3. Santosh Kumar Senapati
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  4. Geeta Rani Jena
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  5. Chinmoy Mishra
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  6. Indramani Nath
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  7. Shantibhusan Senapati
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  8. Manisha Sethi
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Contributions

SKS and IN conceived and designed the study; AS and GRG contributed to the writing and revision of the manuscript; MS performed laboratory testing; AS and CM contributed to the genome sequencing and phylogenetic analysis. SS and MD read and revised the manuscript. All authors’ read and approved the final manuscript.

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Correspondence to Ahmed Magdy Selim.

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Conflict of interest

The authors of this study stated that they have no conflicts of interest.

Ethical standards

There is no precise law for blood sample collection in India for clinical diagnosis and so no consent was obligatory. In this study 4 ml blood collected from the jugular vein under aseptic conditions from cattle after taking the permission of the farm owners.

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Selim, A.M., Das, M., Senapati, S.K. et al. Molecular detection of Theileria annulata infection in cattle by conventional PCR and quantitative real time PCR in India. J Parasit Dis 45, 72–77 (2021). https://doi.org/10.1007/s12639-020-01278-6

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  • DOI: https://doi.org/10.1007/s12639-020-01278-6

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