Presumptive Detection of EHP-Like Structures by Hepatopancreatic Wet Mount in Intensive Pacific White Shrimp Culture
DOI:
https://doi.org/10.30736/grouper.v17i2.497Keywords:
Enterocytozoon hepatopenaei, hepatopancreas, Litopenaeus vannamei, screening, wet mountAbstract
The productivity of Pacific white shrimp (Litopenaeus vannamei) can be constrained by hepatopancreatic microsporidiosis caused by Enterocytozoon hepatopenaei (EHP), a chronic infection associated with growth retardation. This descriptive field case report documented the operational use of unstained hepatopancreatic wet mounts for presumptive microscopic detection of EHP-like structures in two intensive shrimp ponds in Probolinggo Regency, East Java, Indonesia, during October-November 2023. For each pond, 10 shrimp were collected during routine feed-tray inspection for body weight and length assessment, and six individuals were selected for hepatopancreatic wet mount examination at magnifications from 10× to 1,000×. Observations included tubule morphology, lipid droplets, and spore-like structures. The earliest retained observation of EHP-like structures occurred at DOC 19, when the sampled shrimp weighed 0.97 g and measured 5.1 cm. Mean lipid-droplet values were 48.3% in VIP A1 and 29.2% in VVIP A2. EHP-like structures were observed in 3/6 samples (50.0%) from A1 and 5/6 samples (83.3%) from A2. These observations illustrate the potential operational use of wet mounts to select samples requiring confirmatory testing; diagnostic performance remains to be validated against PCR/qPCR or other specific methods.
Downloads
References
Anam, M. K., Ristyanadi, B., & Ubaidillah, M. (2026). CONSUMER PERCEPTION AND SATISFACTION OF BONELESS MILKFISH IN LAMONGAN BASED ON IPA–CSI APPROACH. JP, 16(2), 1018-1031.
Aranguren Caro, L. F., Alghamdi, F., De Belder, K., Lin, J., Mai, H. N., Millabas, J., Alrehaili, Y., Alazwari, A., Algetham, S., & Dhar, A. K. (2021). The effect of salinity on Enterocytozoon hepatopenaei infection in Penaeus vannamei under experimental conditions. BMC Veterinary Research, 17, 65. https://doi.org/10.1186/s12917-021-02778-0
Aranguren Caro, L. F., Mai, H. N., Pichardo, O., Cruz-Flores, R., Hanggono, B., & Dhar, A. K. (2020). Evidences supporting Enterocytozoon hepatopenaei association with white feces syndrome in farmed Penaeus vannamei in Venezuela and Indonesia. Diseases of Aquatic Organisms, 141, 71–78. https://doi.org/10.3354/dao03522
Aras, A. K., Fikriyah, A., Pratiwi, G. A. I., & Nurlita, W. (2022). Prevalensi infeksi EHP (Enterocytozoon hepatopenaei) pada udang vaname (Litopenaeus vannamei) berdasarkan data surveillance di Bali, Indonesia. Media Akuakultur, 17(2), 59–65. https://doi.org/10.15578/ma.17.2.2022.59-65
Cao, Z., Chen, C., Wang, C., Li, T., Chang, L., Si, L., & Yan, D. (2023). Enterocytozoon hepatopenaei (EHP) infection alters the metabolic processes and induces oxidative stress in Penaeus vannamei. Animals, 13(23), 3661. https://doi.org/10.3390/ani13233661
Chaijarasphong, T., Munkongwongsiri, N., Stentiford, G. D., Aldama-Cano, D. J., Thansa, K., Flegel, T. W., Sritunyalucksana, K., & Itsathitphaisarn, O. (2021). The shrimp microsporidian Enterocytozoon hepatopenaei (EHP): Biology, pathology, diagnostics and control. Journal of Invertebrate Pathology, 186, 107458. https://doi.org/10.1016/j.jip.2020.107458
Chen, W.-F., Fu, Y.-W., Zeng, Z.-Y., Guo, S.-Q., Yan, Y.-L., Tu, Y.-F., Gou, T.-G., & Zhang, Q.-Z. (2022). Establishment and application of a TaqMan probe-based qPCR for the detection of Enterocytozoon hepatopenaei in shrimp Litopenaeus vannamei. Parasitology Research, 121(8), 2263–2274. https://doi.org/10.1007/s00436-022-07559-8
Desrina, Prayitno, S. B., Haditomo, A. H. C., Latritiani, R., & Sarjito. (2020). Detection of Enterocytozoon hepatopenaei (EHP) DNA in the polychaetes from shrimp ponds suffering white feces syndrome outbreaks. Biodiversitas, 21(1), 369–374. https://doi.org/10.13057/biodiv/d210144
Duan, Y., Chen, H., Wang, J., Zeng, S., Wang, Y., Mo, Z., Dan, X., & Li, Y. (2021). Response signatures of Litopenaeus vannamei to natural Enterocytozoon hepatopenaei infection revealed by the integration of the microbiome and transcriptome. Aquaculture, 542, 736885. https://doi.org/10.1016/j.aquaculture.2021.736885
Food and Agriculture Organization of the United Nations. (2025, August 29). Shifts in the global shrimp trade and production. GLOBEFISH. https://www.fao.org/in-action/globefish/news-events/news/news-detail/shifts-in-the-global-shrimp-trade-and-production/en
Govindasamy, T., Bhassu, S., & Raju, C. S. (2024). Enterocytozoon hepatopenaei infection in shrimp: Diagnosis, interventions, and food safety guidelines. Microorganisms, 12(1), 21. https://doi.org/10.3390/microorganisms12010021
Kementerian Kelautan dan Perikanan Republik Indonesia. (2026, February 18). Ekspor perikanan tembus USD 6,27 miliar di 2025, KKP: Komoditas Indonesia makin bernilai di pasar global. https://kkp.go.id/news/news-detail/ekspor-perikanan-tembus-usd-627-miliar-di-2025-kkp-komoditas-indonesia-makin-bernilai-di-pasar-global-krB5.html
Khairul, M., Prihatini, E. S., & Mas’ud, F. (2021). Analysis of The Role of Youth Workers in Fishery Sector: Case Study in Lamongan, East Java. Agrikan: Jurnal Agribisnis Perikanan, 14(2), 264-272.
Kumar, T. S., Praveena, P. E., Sivaramakrishnan, T., Rajan, J. J. S., Makesh, M., & Jithendran, K. P. (2022). Effect of Enterocytozoon hepatopenaei (EHP) infection on physiology, metabolism, immunity, and growth of Penaeus vannamei. Aquaculture, 553, 738105. https://doi.org/10.1016/j.aquaculture.2022.738105
Kumar, V., Das, B. K., Dhar, S., Bisai, K., Pande, G. S. J., Zheng, X., Parida, S. N., Adhikari, A., & Jana, A. K. (2024). Ecytonucleospora hepatopenaei (EHP) disease prevalence and mortality in Litopenaeus vannamei: A comparative study from Eastern India shrimp farms. BMC Microbiology, 24, 523. https://doi.org/10.1186/s12866-024-03681-y
Lu, Y.-M., Lu, J.-Q., Zhao, Q., Chen, J., & Xiong, J.-B. (2025). Pathogenic mechanisms of Enterocytozoon hepatopenaei through the parasite–gut microbiome–shrimp (Litopenaeus vannamei) physiology axis. Zoological Research, 46(2), 401–413. https://doi.org/10.24272/j.issn.2095-8137.2024.411
López-Carvallo, J. A., Cruz-Flores, R., & Dhar, A. K. (2022). The emerging pathogen Enterocytozoon hepatopenaei drives a degenerative cyclic pattern in the hepatopancreas microbiome of the shrimp (Penaeus vannamei). Scientific Reports, 12, 14766. https://doi.org/10.1038/s41598-022-19127-2
Ma, C., Fan, S., Wang, Y., Yang, H., Qiao, Y., Jiang, G., Lyu, M., Dong, J., Shen, H., & Gao, S. (2021). Rapid detection of Enterocytozoon hepatopenaei infection in shrimp with a real-time isothermal recombinase polymerase amplification assay. Frontiers in Cellular and Infection Microbiology, 11, 631960. https://doi.org/10.3389/fcimb.2021.631960
Nkuba, A. C., Mahasri, G., Lastuti, N. D. R., & Mwendolwa, A. A. (2021). Correlation of nitrite and ammonia with prevalence of Enterocytozoon hepatopenaei (EHP) in shrimp (Litopenaeus vannamei) on several super-intensive ponds in East Java, Indonesia. Jurnal Ilmiah Perikanan dan Kelautan, 13(1), 58–67. https://doi.org/10.20473/jipk.v13i1.24430
Patil, P. K., Geetha, R., Ravisankar, T., Avunje, S., Solanki, H. G., Abraham, T. J., Vinoth, S. P., Jithendran, K. P., Alavandi, S. V., & Vijayan, K. K. (2021). Economic loss due to diseases in Indian shrimp farming with special reference to Enterocytozoon hepatopenaei (EHP) and white spot syndrome virus (WSSV). Aquaculture, 533, 736231. https://doi.org/10.1016/j.aquaculture.2020.736231
Piamsomboon, P., & Han, J. E. (2022). White feces syndrome, a multifactorial syndrome of cultured shrimp: A mini review. Fishes, 7(6), 339. https://doi.org/10.3390/fishes7060339
Raj, M., Sathiyaraj, G., Narayanan, B., Babu, B., Varkey, M., Karthickkannan, K., Ganeshamurthy, R., Mandal, A., & Kandan, S. (2023). Characterization of Enterocytozoon hepatopenaei causing hepatopancreatic microsporidiosis in L. vannamei and a new molecular method for its detection in shrimps, and other environmental samples. Journal of Invertebrate Pathology, 199, 107951. https://doi.org/10.1016/j.jip.2023.107951
Villarreal, H., & Juarez, L. M. (2022). Super-intensive shrimp culture: Analysis and future challenges. Journal of the World Aquaculture Society, 53(5), 928–932. https://doi.org/10.1111/jwas.12929
Virapat, C. (2023). The current challenges of sustainable aquaculture in the Asia-Pacific Region and the measures recommended. Asia-Pacific Journal of Rural Development, 32(2), 207–216. https://doi.org/10.1177/10185291231193516
Wan Sajiri, W. M. H., Kua, B. C., & Borkhanuddin, M. H. (2023). Detection of Enterocytozoon hepatopenaei (EHP) (microsporidia) in several species of potential macrofauna-carriers from shrimp (Penaeus vannamei) ponds in Malaysia. Journal of Invertebrate Pathology, 198, 107910. https://doi.org/10.1016/j.jip.2023.107910
Wang, L., Lv, Q., He, Y., Gu, R., Zhou, B., Chen, J., Fan, X., Pan, G., Long, M., & Zhou, Z. (2020). Integrated qPCR and staining methods for detection and quantification of Enterocytozoon hepatopenaei in shrimp Litopenaeus vannamei. Microorganisms, 8(9), 1366. https://doi.org/10.3390/microorganisms8091366
Wang, Y., Chen, J., Na, Y., Li, X.-C., Zhou, J.-F., Fang, W.-H., & Tan, H.-X. (2023). Ecytonucleospora hepatopenaei n. gen. et comb. (Microsporidia: Enterocytozoonidae): A redescription of Enterocytozoon hepatopenaei (Tourtip et al., 2009), a microsporidian infecting the widely cultivated shrimp Penaeus vannamei. Journal of Invertebrate Pathology, 201, 107988.
Wu, Y., Chen, J., Liao, G., Hu, M., Zhang, Q., Meng, X., Li, T., Long, M., Fan, X., Yu, Q., Zhang, L., Pan, G., & Zhou, Z. (2022). Down-regulation of lipid metabolism in the hepatopancreas of shrimp Litopenaeus vannamei upon light and heavy infection of Enterocytozoon hepatopenaei: A comparative proteomic study. International Journal of Molecular Sciences, 23(19), 11574. https://doi.org/10.3390/ijms231911574
Yuanlae, S., Thaiue, D., Saedan, S., Kittiwongpukdee, K., Vanichviriyakit, R., Chuchird, N., & Itsathitphaisarn, O. (2025). Ecytonucleospora hepatopenaei causes lipid droplet depletion and imbalanced lipid metabolism in Penaeus vannamei. Scientific Reports, 15, 37198. https://doi.org/10.1038/s41598-025-21037-y
Zhao, R.-H., Gao, W., Qiu, L., Chen, X., Dong, X., Li, C., & Huang, J. (2020). A staining method for detection of Enterocytozoon hepatopenaei (EHP) spores with calcofluor white. Journal of Invertebrate Pathology, 172, 107347. https://doi.org/10.1016/j.jip.2020.107347
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2026 Hanif Nadhifa Esrarf, Laksmi Sulmartiwi, Abdul Manan
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
.jpg){kind=logo}
1.png){kind=logo}
