Molecular Detection and Phylogeny of Hepatozoon Ophisauri and Toxoplasma Gondii in Wild Lizards from Khyber Pakhtunkhwa, Pakistan

Overview

Journal Folia Microbiol (Praha)

Publisher Springer

Specialty Microbiology

Date 2025 Feb 22

PMID 39985744

Authors

Sidra Ghafar

Asmat Ullah Khan

Mustansar Khalid

Sami Ul Haq

Saeed Ullah

Umaima Basit

Hira Muqaddas

Ibrahim Abbas Mohammed

Ayat Ammar Mahdi

Adil Khan

Mourad Ben Said

Furhan Iqbal

Affiliations

Soon will be listed here.

Abstract

Despite Pakistan's rich reptilian diversity, wild lizards have largely been unexplored for vector-borne parasites. This study reports the prevalence and phylogenetic assessment of Hepatozoon ophisauri and Toxoplasma gondii DNA in the blood samples of 101 wild lizards from Khyber Pakhtunkhwa, Pakistan, captured at altitudes of 1200 to 2250 m above sea level between March 2022 and June 2023. PCR-based molecular analysis identified H. ophisauri DNA in three (3%) lizards of all Laudakia tuberculata. Additionally, 45 (44.5%) lizards tested positive for T. gondii DNA, including L. agrorensis, L. pakistanica, L. tuberculata, and Abblepharus pannonicus. Sequence analysis and BLAST confirmed the presence of H. ophisauri and T. gondii. Phylogenetic analysis showed genetic diversity among the H. ophisauri and T. gondii nucleotide sequences, clustering with reference sequences from reptiles, birds, and ticks. Toxoplasma gondii prevalence varied among species, with the highest rates in L. agrorensis and A. pannonicus (67%), followed by L. pakistanica (45%) and L. tuberculata (43%). This study is the first from Pakistan to report high T. gondii prevalence and low H. ophisauri prevalence in wild lizards. Similar and larger-scale studies in unexplored regions of Pakistan are needed to enhance understanding of these pathogens' genetic diversity and host-parasite interactions.

References

Abdel-Baki A, Abdel-Haleem H, Abdel-Tawab H, Al-Quraishy S, Mansour L . Reassignment of Haemogregarina annularis from the Blood of Tarentola annularis to the Genus Hepatozoon Based on the Parasite Morphology and 18S rDNA Sequence Analysis. Acta Parasitol. 2021; 67(1):411-417. DOI: 10.1007/s11686-021-00477-2. View

Ahmad G, Masud A, Naeem M, Ghafar A, Muqaddas H, Qamar M . Molecular prevalence, phylogeny and hematological impact of Toxoplasma gondii and Plasmodium spp. in common quails from Punjab, Pakistan. PLoS One. 2024; 19(5):e0304179. PMC: 11142681. DOI: 10.1371/journal.pone.0304179. View

Aktas M, Ozubek S, Sayin Ipek D . Molecular investigations of Hepatozoon species in dogs and developmental stages of Rhipicephalus sanguineus. Parasitol Res. 2013; 112(6):2381-5. DOI: 10.1007/s00436-013-3403-6. View

Aktas M, Ozubek S, Altay K, Balkaya I, Utuk A, Kirbas A . A molecular and parasitological survey of Hepatozoon canis in domestic dogs in Turkey. Vet Parasitol. 2015; 209(3-4):264-7. DOI: 10.1016/j.vetpar.2015.02.015. View

Borges-Nojosa D, Borges-Leite M, Maia J, Zanchi-Silva D, da Rocha Braga R, Harris D . A new species of Hepatozoon Miller, 1908 (Apicomplexa: Adelerina) from the snake Philodryas nattereri Steindachner (Squamata: Dipsadidae) in northeastern Brazil. Syst Parasitol. 2017; 94(1):65-72. DOI: 10.1007/s11230-016-9676-2. View

de Camps S, Dubey J, Saville W . Seroepidemiology of Toxoplasma gondii in zoo animals in selected zoos in the midwestern United States. J Parasitol. 2008; 94(3):648-53. DOI: 10.1645/GE-1453.1. View

Er-Rguibi O, Harris D, Aglagane A, Laghzaoui E, Kimdil L, Abbad A . Molecular Survey and Microscopic Examination of Haemoparasites Infecting Lizards from Morocco. Acta Parasitol. 2023; 68(3):593-603. DOI: 10.1007/s11686-023-00688-9. View

Godfrey S, Nelson N, Bull C . Ecology and dynamics of the blood parasite, Hepatozoon tuatarae (Apicomplexa), in tuatara (Sphenodon punctatus) on Stephens Island, New Zealand. J Wildl Dis. 2011; 47(1):126-39. DOI: 10.7589/0090-3558-47.1.126. View

Goncalves I, Uzeda R, Lacerda G, Moreira R, Araujo F, Oliveira R . Molecular frequency and isolation of cyst-forming coccidia from free ranging chickens in Bahia State, Brazil. Vet Parasitol. 2012; 190(1-2):74-9. DOI: 10.1016/j.vetpar.2012.05.007. View

10.

Gondim L, Abe-Sandes K, Uzeda R, Silva M, Santos S, Mota R . Toxoplasma gondii and Neospora caninum in sparrows (Passer domesticus) in the Northeast of Brazil. Vet Parasitol. 2009; 168(1-2):121-4. DOI: 10.1016/j.vetpar.2009.09.055. View

11.

Harris D, Maia J, Perera A . Molecular survey of Apicomplexa in Podarcis wall lizards detects Hepatozoon, Sarcocystis, and Eimeria species. J Parasitol. 2012; 98(3):592-7. DOI: 10.1645/JP-GE-2908.1. View

12.

Magnino S, Colin P, Dei-Cas E, Madsen M, McLauchlin J, Nockler K . Biological risks associated with consumption of reptile products. Int J Food Microbiol. 2009; 134(3):163-75. DOI: 10.1016/j.ijfoodmicro.2009.07.001. View

13.

Maia J, Perera A, Harris D . Molecular survey and microscopic examination of Hepatozoon Miller, 1908 (Apicomplexa: Adeleorina) in lacertid lizards from the western Mediterranean. Folia Parasitol (Praha). 2013; 59(4):241-8. DOI: 10.14411/fp.2012.033. View

14.

Nasiri V, Teymurzadeh S, Karimi G, Nasiri M . Molecular detection of Toxoplasma gondii in snakes. Exp Parasitol. 2016; 169:102-6. DOI: 10.1016/j.exppara.2016.08.002. View

15.

ODwyer L, Moco T, Paduan K, Spenassatto C, Silva R, Ribolla P . Description of three new species of Hepatozoon (Apicomplexa, Hepatozoidae) from Rattlesnakes (Crotalus durissus terrificus) based on molecular, morphometric and morphologic characters. Exp Parasitol. 2013; 135(2):200-7. DOI: 10.1016/j.exppara.2013.06.019. View

16.

Swei A, Ostfeld R, Lane R, Briggs C . Impact of the experimental removal of lizards on Lyme disease risk. Proc Biol Sci. 2011; 278(1720):2970-8. PMC: 3151702. DOI: 10.1098/rspb.2010.2402. View

17.

Tippery N, Les D . Phylogenetic analysis of the internal transcribed spacer (ITS) region in Menyanthaceae using predicted secondary structure. Mol Phylogenet Evol. 2008; 49(2):526-37. DOI: 10.1016/j.ympev.2008.07.019. View

18.

Tome B, Maia J, Harris D . Molecular assessment of apicomplexan parasites in the snake Psammophis from North Africa: do multiple parasite lineages reflect the final vertebrate host diet?. J Parasitol. 2013; 99(5):883-7. DOI: 10.1645/12-95.1. View

19.

Wozniak E, Telford Jr S . The fate of Hepatozoon species naturally infecting Florida black racers and watersnakes in potential mosquito and soft tick vectors, and histological evidence of pathogenicity in unnatural host species. Int J Parasitol. 1991; 21(5):511-6. DOI: 10.1016/0020-7519(91)90055-c. View

20.

Zechmeisterova K, Pribyl M, Nguyen H, Noskova E, Siroky P . Haemogregarines of the Genera Haemogregarina, Hemolivia, and Hepatozoon Infecting Vietnamese Freshwater Turtles, with Additional Notes on Primer Specificity and Primer-template Mismatches Affecting Diagnostic Success. Protist. 2022; 173(4):125884. DOI: 10.1016/j.protis.2022.125884. View