Molecular detection and identification of Plasmodium spp. isolated from captive-bred cynomolgus monkeys in Bogor, Indonesia

Vet World Vol.17 February-2024 Article-11

Research Article

Veterinary World, 17(2): 337-343

https://doi.org/10.14202/vetworld.2024.337-343

Uus Saepuloh¹, Lis Rosmanah¹, Risqa Novita^2,3, Ellis Dwi Ayuningsih¹, Susi Soviana⁴, Upik Kesumawati Hadi⁴, and Huda Shalahudin Darusman^1,4

1. Primate Research Center, Bogor Agricultural University, Jl. Lodaya II/5, Bogor, 16151, Indonesia.
2. Research Center for Pharmaceutical Ingredients and Traditional Medicine, National Research and Innovation Agency (BRIN), Genomic Building, Cibinong Science Center, Jl. Raya Bogor No. 490, Cibinong, 16915 Indonesia.
3. Primatology Study Program, Graduate School of IPB University, Jl. Lodaya II/5, Bogor, 16151, Indonesia.
4. Department of Animal Infectious Diseases and Veterinary Public Health, Faculty of Veterinary Medicine, Bogor Agricultural University, Jl. Agatis, Dramaga, Bogor, 16680, Indonesia.

Background and Aim: Asian macaques are natural hosts of several Plasmodium species. Some monkey malaria parasites may infect humans and cause zoonotic infections. This study was conducted to estimate the prevalence of monkey malaria parasites in Bogor, Indonesia, based on molecular detection and identification, particularly in cynomolgus monkeys, which have a wide geographic distribution and share extensive habitats with humans. These data are needed to evaluate the status of simian malaria among macaques in Bogor and to study the potential risks to human health. These updated data will provide sufficient information for implementing malaria control strategies in the future and for developing a potential malaria vaccine using monkeys as an animal model.

Materials and Methods: Blood samples of 274 cynomolgus monkeys (Macaca fascicularis) were collected and identified using microscopy. DNA was extracted from positive blood samples and analyzed using polymerase chain reaction (PCR) to amplify the small subunit ribosomal RNA (SSU rRNA) target gene using consensus primers for Plasmodium species. The PCR-positive samples were then nucleotide-sequenced using commercial sequencing services, analyzed using the BioEdit program, and aligned using Basic Local Alignment Search Tool from the National Center for Biotechnology Information. Phylogenetic trees were constructed using MEGA 11.0 and the neighbor-joining (NJ) method to determine the kinship of Plasmodium. Bootstrapping was performed using 500 replicates to assess the robustness of tree topologies.

Results: Thirty-eight of the 274 microscopically positive samples for Plasmodium spp. were also positive using PCR, resulting in a 1640 bp amplicon. Further, analysis using nucleotide sequencing confirmed that these positive samples were Plasmodium inui with more than 99% nucleotide identity compared to GenBank sequences. Phylogenetic tree analysis of the SSU rRNA partial gene showed that all our isolates clustered and were closely related to a P. inui strain isolated from cynomolgus macaques in South China in 2011.

Conclusion: P. inui is the predominant malaria parasite in cynomolgus monkeys from Bogor. Keywords: malaria, phylogenetic tree, Plasmodium inui, small subunit ribosomal RNA.

Keywords: malaria, phylogenetic tree, Plasmodium inui, small subunit ribosomal RNA.

How to cite this article: Saepuloh U, Rosmanah L, Novita R, Ayuningsih ED, Soviana S, Hadi UK, and Darusman HS (2024) Molecular detection and identification of Plasmodium spp. isolated from captive-bred cynomolgus monkeys in Bogor, Indonesia, Veterinary World, 17(2): 337-343.

Received: 03-09-2023 Accepted: 15-01-2024 Published online: 08-02-2024

Corresponding author: Huda Shalahudin Darusman E-mail: hudada@apps.ipb.ac.id

DOI: 10.14202/vetworld.2024.337-343

Copyright: Saepuloh, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.