First molecular evidence of Hepatocystis infection in non-human primates from Indonesia using fecal DNA: Implications for wildlife surveillance and One Health.
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| Title: | First molecular evidence of Hepatocystis infection in non-human primates from Indonesia using fecal DNA: Implications for wildlife surveillance and One Health. |
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| Authors: | Hasan, Firmanul1 hasanfirmanul@gmail.com, Siregar, Josephine Elizabeth2 jose001@brin.go.id, Pravitasari, Normalita Eka3 normalitapravitasari@yahoo.com, Rizki, Andita Fitri Mutiara2 anditafitri.mutiara@gmail.com, Azmi, Wihda Aisarul2 wihda.aiss@gmail.com, Artika, I. Made1 imart@apps.ipb.ac.id, Kuswanda, Wanda4 wand002@brin.go.id |
| Source: | Veterinary World. Nov2025, Vol. 18 Issue 11, p3651-3669. 19p. |
| Document Type: | Article |
| Subjects: | Primates, Molecular diagnosis, Zoonoses, Wildlife monitoring, Blood parasites, Indonesians, Environmental health |
| Geographic Terms: | Indonesia |
| Author-Supplied Keywords: | fecal DNA Hepatocystis non-human primates One Health Plasmodium small subunit ribosomal RNA |
| Abstract: | Background and Aim: Hepatocystis, a malaria-like hemoparasite closely related to Plasmodium, infects non-human primates (NHPs), bats, and other mammals, yet remains understudied in Indonesia. Although Plasmodium detection in primates has been extensively reported, molecular confirmation of Hepatocystis in Indonesian wildlife is lacking. This study aimed to screen NHP fecal samples for Plasmodium infection and to identify any malaria-like parasites using molecular methods. Materials and Methods: A total of 227 fecal samples from captive and rescued NHPs, representing multiple Macaca species and other primates, were collected from Tasikoki Wildlife Rescue Center, Manado, Indonesia, in 2019 and 2021. Genomic DNA was extracted using a QIAamp Fast DNA Stool Mini Kit (Qiagen, Germany) and screened for Plasmodium using polymerase chain reaction (PCR) targeting the mitochondrial small subunit ribosomal RNA gene. Positive amplicons were purified, sequenced, and analyzed using the basic local alignment search tool and phylogenetic reconstruction with MEGA X. Results: Eight (3.5%) of 227 samples yielded positive PCR bands of approximately 600 bp, differing from the expected 467 bp for Plasmodium. Sequencing of four representative samples (MNig-01, MNig-17, MNig-18, and HM-160) revealed >99.7% identity with Hepatocystis spp. (GenBank: KY653782.1). Multiple sequence alignment confirmed complete nucleotide conservation among the four isolates, and phylogenetic analysis clustered them within the Hepatocystis clade, closely related to Hepatocystis spp. ex Pteropus hypomelanus from Malaysia. All positive detections were from 2019 samples, suggesting temporal variation in infection or vector activity. Conclusion: This study provides the first molecular evidence of Hepatocystis infection in Indonesian NHPs using fecal DNA, expanding current knowledge of parasite distribution and host range. The successful detection of Hepatocystis through non-invasive sampling underscores the potential of fecal-based PCR for wildlife disease surveillance. These findings highlight the importance of integrating molecular diagnostics into conservation and One Health frameworks to monitor zoonotic parasites and understand host–vector–pathogen interactions in natural ecosystems. [ABSTRACT FROM AUTHOR] |
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| Author Affiliations: | 1Department of Biochemistry, Faculty of Mathematics and Natural Sciences, IPB University, Dramaga Campus, Bogor 16680, Indonesia. 2Eijkman Research Center for Molecular Biology, National Research and Innovation Agency, Cibinong, Bogor 16911, Indonesia. 3School of Tropical Medicine and Global Health, Nagasaki University, Nagasaki City, Japan. 4Research Center for Biota Systems, National Research and Innovation Agency, Cibinong, Bogor 16911, Indonesia. |
| ISSN: | 0972-8988 |
| DOI: | 10.14202/vetworld.2025.3651-3669 |
| Accession Number: | 189867715 |
| Database: | Veterinary Source |
| Abstract: | Background and Aim: Hepatocystis, a malaria-like hemoparasite closely related to Plasmodium, infects non-human primates (NHPs), bats, and other mammals, yet remains understudied in Indonesia. Although Plasmodium detection in primates has been extensively reported, molecular confirmation of Hepatocystis in Indonesian wildlife is lacking. This study aimed to screen NHP fecal samples for Plasmodium infection and to identify any malaria-like parasites using molecular methods. Materials and Methods: A total of 227 fecal samples from captive and rescued NHPs, representing multiple Macaca species and other primates, were collected from Tasikoki Wildlife Rescue Center, Manado, Indonesia, in 2019 and 2021. Genomic DNA was extracted using a QIAamp Fast DNA Stool Mini Kit (Qiagen, Germany) and screened for Plasmodium using polymerase chain reaction (PCR) targeting the mitochondrial small subunit ribosomal RNA gene. Positive amplicons were purified, sequenced, and analyzed using the basic local alignment search tool and phylogenetic reconstruction with MEGA X. Results: Eight (3.5%) of 227 samples yielded positive PCR bands of approximately 600 bp, differing from the expected 467 bp for Plasmodium. Sequencing of four representative samples (MNig-01, MNig-17, MNig-18, and HM-160) revealed >99.7% identity with Hepatocystis spp. (GenBank: KY653782.1). Multiple sequence alignment confirmed complete nucleotide conservation among the four isolates, and phylogenetic analysis clustered them within the Hepatocystis clade, closely related to Hepatocystis spp. ex Pteropus hypomelanus from Malaysia. All positive detections were from 2019 samples, suggesting temporal variation in infection or vector activity. Conclusion: This study provides the first molecular evidence of Hepatocystis infection in Indonesian NHPs using fecal DNA, expanding current knowledge of parasite distribution and host range. The successful detection of Hepatocystis through non-invasive sampling underscores the potential of fecal-based PCR for wildlife disease surveillance. These findings highlight the importance of integrating molecular diagnostics into conservation and One Health frameworks to monitor zoonotic parasites and understand host–vector–pathogen interactions in natural ecosystems. [ABSTRACT FROM AUTHOR] |
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| ISSN: | 09728988 |
| DOI: | 10.14202/vetworld.2025.3651-3669 |