Skip to main content
SpringerLink
Log in

Phylogenetic analysis and haplotype networking of Hepatozoon felis infecting wild animals in Gir National Park, Gujarat, India

  • Research
  • Published:
Parasitology Research Aims and scope Submit manuscript

Abstract

The present study was aimed to determine the phylogenetic relationship, haplotype network, and demographic dynamics of H. felis infecting the endangered Asiatic lions in Gir National Park, Gujarat, India, on the basis of partial 18S rRNA gene. The phylogenetic analysis based on the partial 18S rRNA gene sequences of H. felis exhibited the presence of two distinct genotypes of H. felis (HfG1 and HfG2) infecting the Indian wild felids, viz., the Asiatic lion, Royal Bengal tiger, and Indian leopard. The HfG1 and HfG2 genotypes exhibited 97.6–100% and 99.7–100%, and 96.9–98.7% nucleotide identity within and between themselves, respectively. The HfG1 genotype exhibited a higher genetic diversity as compared to HfG2. A total of 22 molecular signatures were identified in the 18S rRNA gene between these genotypes. Further, analysis of a total 67 sequences of H. felis (13 different host species from 13 countries of Africa, South America, Asia, and Europe) that were downloaded from GenBankTM, generated 30 haplotypes. Among all the haplotypes, Hap_17 (h=12) was the most frequent followed by Hap_12 (h=09) and Hap_4 (h=05). Out of 13 location-wise populations, India (h=12) shared the highest number of haplotypes followed by Japan (h=08), and the least number of haplotypes were found in Hungary (h=02). Population dynamics study involving neutrality tests and mismatch distribution, and genetic differentiation indices, revealed the presence of phylogeographic population structure and a constant population size indicating a uniform gene flow among the populations worldwide. In conclusion, a high genetic diversity along with the presence of two distinct genotypes of H. felis were observed on the basis of 18S rRNA gene sequence analysis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Data availability

All the nucleotide sequences generated and used in the present study are available in the GenBankTM. The data supporting the findings of this study are available within the article.

References

  • Attipa C, Neofytou K, Yiapanis C, Martínez-Orellana P, Baneth G, Nachum-Biala Y, Brooks-Brownlie H, Solano-Gallego L, Tasker S (2017) Follow-up monitoring in a cat with leishmaniosis and coinfections with Hepatozoon felis and ‘Candidatus Mycoplasma haemominutum’. J Feline Med Surg 3(2):2055116917740454

    Google Scholar 

  • Baneth G, Sheiner A, Eyal O, Hahn S, Beaufils JP, Anug Y, Talmi-Frank D (2013) Redescription of Hepatozoon felis (Apicomplexa: Hepatozoidae) based on phylogenetic analysis, tissue and blood form morphology, and possible transplacental transmission. Parasit Vectors 6(1):1–10

    Article  Google Scholar 

  • Bhusri B, Sariya L, Mongkolphan C, Suksai P, Kaewchot S, Changbunjong T (2017) Molecular characterization of Hepatozoon felis in Rhipicephalus sanguineus ticks infested on captive lions (Panthera leo). J Parasit Dis 41(3):903–907

    Article  PubMed  PubMed Central  Google Scholar 

  • Chauhan RP, Kumari A, Nehra AK, Ram H, Garg R, Banerjee PS, Karikalan M, Sharma AK (2020) Genetic characterization and phylogenetic analysis of Sarcocystis suihominis infecting domestic pigs (Sus scrofa) in India. Parasitol Res 119:3347–3357

    Article  PubMed  Google Scholar 

  • Criado-Fornelio A, Ruas JL, Casado N, Farias NAR, Soares MP, Müller G, Brum JGW, Berne MEA, Buling-Saraña A, Barba-Carretero JC (2006) New molecular data on mammalian Hepatozoon species (Apicomplexa: Adeleorina) from Brazil and Spain. J Parasitol 92(1):93–99

    Article  CAS  PubMed  Google Scholar 

  • de Bortoli CP, André MR, Braga MDSC, Machado RZ (2011) Molecular characterization of Hepatozoon sp. in cats from São Luís Island, Maranhão, northeastern Brazil. Parasitol Res 109(4):1189–1192

    Article  PubMed  Google Scholar 

  • Giannelli A, Latrofa MS, Nachum-Biala Y, Hodžić A, Greco G, Attanasi A, Annoscia G, Otranto D, Baneth G (2017) Three different Hepatozoon species in domestic cats from southern Italy. Ticks Tick Borne Dis 8(5):721–724

    Article  PubMed  Google Scholar 

  • Grillini M, Simonato G, Tessarin C, Dotto G, Traversa D, Cassini R, Marchiori E, Frangipane di Regalbono A (2021) Cytauxzoon sp. and Hepatozoon spp. in domestic cats: A preliminary study in North-Eastern Italy. Pathogens 10(9):1214

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Harris DJ, Santos J, Rampedi KM, Halajian A, Xavier R (2019) Genetic diversity of Hepatozoon (Apicomplexa) from domestic cats in South Africa, with a global reassessment of Hepatozoon felis diversity. J S Afr Vet Assoc 90(1):1–6

    Google Scholar 

  • Hodžić A, Alić A, Prašović S, Otranto D, Baneth G, Duscher GG (2017) Hepatozoon silvestris sp. nov.: morphological and molecular characterization of a new species of Hepatozoon (Adeleorina: Hepatozoidae) from the European wild cat (Felis silvestris silvestris). Parasitology 144(5):650–661

    Article  PubMed  Google Scholar 

  • Hornok S, Boldogh SA, Takács N, Kontschán J, Szekeres S, Sós E, Sándor AD, Wang Y, Tuska-Szalay B (2022) Molecular epidemiological study on ticks and tick-borne protozoan parasites (Apicomplexa: Cytauxzoon and Hepatozoon spp.) from wild cats (Felis silvestris), Mustelidae and red squirrels (Sciurus vulgaris) in central Europe. Hungary. Parasit Vectors 15(1):1–11

    Google Scholar 

  • Hudson RR, Slatkin M, Maddison WP (1992) Estimation of levels of gene flow from DNA sequence data. Genetics 132(2):583–589

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ikadai H, Tanaka H, Shibahara N, Matsuu A, Uechi M, Itoh N, Oshiro S, Kudo N, Igarashi I, Oyamada T (2004) Molecular evidence of infections with Babesia gibsoni parasites in Japan and evaluation of the diagnostic potential of a loop-mediated isothermal amplification method. J Clin Microbiol 42(6):2465–2469

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kubo M, Jeong A, Kim SI, Kim YJ, Lee H, Kimura J, Agatsuma T, Sakai H, Yanai T (2010) The first report of Hepatozoon species infection in leopard cats (Prionailurus bengalensis) in Korea. J Parasitol 96(2):437–439

    Article  PubMed  Google Scholar 

  • Kundave VR, Ram H, Shahzad M, Garg R, Banerjee PS, Nehra AK, Rafiqi SI, Ravikumar G, Tiwari AK (2019) Genetic characterization of Theileria species infecting bovines in India. Infect Genet Evol 75:103962

    Article  CAS  PubMed  Google Scholar 

  • Leigh JW, Bryant D (2015) POPART: full-feature software for haplotype network construction. Methods Ecol Evol 6(9):1110–1116

    Article  Google Scholar 

  • Lloret A, Addie DD, Boucraut-Baralon C, Egberink H, Frymus T, Gruffydd-Jones T, Hartmann K, Horzinek MC, Hosie MJ, Lutz H, Marsilio F (2015) Hepatozoonosis in cats: ABCD guidelines on prevention and management. J Feline Med Surg 17(7):642–644

    Article  PubMed  Google Scholar 

  • Lynch M, Crease TJ (1990) The analysis of population survey data on DNA sequence variation. Mol Biol Evol 7:377–394

    CAS  PubMed  Google Scholar 

  • Mandal M, Banerjee PS, Garg R, Ram H, Kundu K, Kumar S, Kumar GR (2014) Genetic characterization and phylogenetic relationships based on 18S rRNA and ITS1 region of small form of canine Babesia spp. from India. Infect Genet Evol 27:325–331

    Article  CAS  PubMed  Google Scholar 

  • Morelli S, Diakou A, Traversa D, Di Gennaro E, Simonato G, Colombo M, Dimzas D, Grillini M, di Regalbono AF, Beugnet F, Halos L (2021) First record of Hepatozoon spp. in domestic cats in Greece. Ticks Tick Borne Dis 12(1):101580

    Article  PubMed  Google Scholar 

  • Nehra AK, Kumari A, Moudgil AD, Vohra S (2021) Ticks Tick Borne Dis. 12:101776. https://doi.org/10.1016/j.ttbdis.2021.101776

    Article  PubMed  Google Scholar 

  • Nehra AK, Kumari A, Kundave VR, Vohra S, Ram H (2022) Molecular insights into the population structure and haplotype network of Theileria annulata based on the small-subunit ribosomal RNA (18S rRNA) gene. Infect Genet Evol 99:105252

    Article  CAS  PubMed  Google Scholar 

  • Nehra AK, Kumari A, Moudgil AD, Vohra S (2022) An insight into misidentification of the small-subunit ribosomal RNA (18S rRNA) gene sequences of Theileria spp. as Theileria annulata. BMC Vet Res 18(1):1–9

    Article  Google Scholar 

  • Orkun Ö (2022) Description of a novel Babesia sp. genotype from a naturally infected Eurasian lynx (Lynx lynx) in Anatolia, Turkey, with remarks on its morphology and phylogenetic relation to other piroplasmid species. Ticks Tick Borne Dis 13(6):102026

    Article  PubMed  Google Scholar 

  • Ortuño A, Castellà J, Criado-Fornelio A, Buling A, Barba-Carretero JC (2008) Molecular detection of a Hepatozoon species in stray cats from a feline colony in North-eastern Spain. Vet J 177(1):134–135

    Article  PubMed  Google Scholar 

  • Patton WS (1908) The haemogregarines of mammals and reptiles. Parasitol 1(4):318–321

    Article  Google Scholar 

  • Pawar RM, Poornachandar A, Srinivas P, Rao KR, Lakshmikantan U, Shivaji S (2012) Molecular characterization of Hepatozoon spp infection in endangered Indian wild felids and canids. Vet Parasitol 186(3–4):475–9

    Article  CAS  PubMed  Google Scholar 

  • Rafiqi SI, Kumar S, Reena KK, Garg R, Ram H, Karikalan M, Mahendran K, Pawde AM, Sharma AK, Banerjee PS (2018) Molecular characterization of Hepatozoon sp. and Babesia sp. isolated from endangered Asiatic lion (Panthera leo persica). Indian J Anim Sci 88:662–666

    Article  CAS  Google Scholar 

  • Rozas J, Ferrer-Mata A, Sánchez-DelBarrio JC, Guirao-Rico S, Librado P, Ramos-Onsins SE, Sánchez-Gracia A (2017) DnaSP 6: DNA sequence polymorphism analysis of large data sets. Mol Biol Evol 34(12):3299–3302

    Article  CAS  PubMed  Google Scholar 

  • Tamura K (1992) Estimation of the number of nucleotide substitutions when there are strong transition-transversion and G+ C-content biases. Mol Biol Evol 9(4):678–687

    CAS  PubMed  Google Scholar 

  • Tamura K, Stecher G, Kumar S (2021) MEGA 11: Molecular Evolutionary Genetics Analysis Version 11. Mol Biol Evol. https://doi.org/10.1093/molbev/msab120

    Article  PubMed  PubMed Central  Google Scholar 

  • Vásquez-Aguilar AA, Barbachano-Guerrero A, Angulo DF, Jarquín-Díaz VH (2021) Phylogeography and population differentiation in Hepatozoon canis (Apicomplexa: Hepatozoidae) reveal expansion and gene flow in world populations. Parasit Vectors 14(1):1–14

    Article  Google Scholar 

  • Venkatesan T, Panda R, Kumari A, Nehra AK, Ram H, Pateer DP, Karikalan M, Garg R, Singh MK, Shukla U, Pawde AM (2022) Genetic and population diversity of Toxocara cati (Schrank, 1788) Brumpt, 1927, on the basis of the internal transcribed spacer (ITS) region. Parasitol Res 121(12):3477–3493

    Article  PubMed  Google Scholar 

Download references

Acknowledgements

The authors are thankful to the Director, ICAR-IVRI, Izatnagar, Uttar Pradesh (India), for providing the necessary facilities to carry out this research work.

Funding

This work was funded by ICAR- Indian Veterinary Research Institute, Izatnagar (U.P.), India.

Author information

Authors and Affiliations

  1. Division of Parasitology, ICAR- Indian Veterinary Research Institute, Izatnagar (U.P.), India

    Rasmita Panda, Anil Kumar Nehra, Hira Ram & Rajat Garg

  2. Department of Veterinary Parasitology, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar (Haryana), India

    Anil Kumar Nehra

  3. Centre for Wildlife Conservation, Management and Disease Surveillance, ICAR- Indian Veterinary Research Institute, Izatnagar (U.P.), India

    Mathesh Karikalan & A. M. Pawde

  4. Sardar Patel Zoological Park, Kevadia, Gujarat, India

    Ram Ratan Nala

Authors
  1. Rasmita Panda
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Anil Kumar Nehra
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hira Ram
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mathesh Karikalan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Rajat Garg
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ram Ratan Nala
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. A. M. Pawde
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Rasmita Panda: Original draft preparation, Investigation, Methodology.

Anil Kumar Nehra: Conceptualization, Methodology, Software, Formal analysis, Writing - review and editing.

Hira Ram: Writing - review and editing; Supervision.

Mathesh Karikalan: Investigation.

Ram Ratan Nala: Investigation.

Rajat Garg, A.M. Pawde: Writing - review and editing.

Corresponding author

Correspondence to Hira Ram.

Ethics declarations

Competing interests

The authors declare no competing interests.

Ethical approval

Not applicable.

Consent to participate

Not applicable.

Consent for publication

Not applicable.

Competing interests

The authors declare no competing interests.

Additional information

Section Editor: Otranto Domenico

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rasmita Panda and Anil Kumar Nehra are equal first authors.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 5.76 MB)

Supplementary file2 (DOCX 19 KB)

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Panda, R., Nehra, A.K., Ram, H. et al. Phylogenetic analysis and haplotype networking of Hepatozoon felis infecting wild animals in Gir National Park, Gujarat, India. Parasitol Res 123, 92 (2024). https://doi.org/10.1007/s00436-023-08109-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s00436-023-08109-6

Keywords

Search

Navigation