Correlation of Nitrite and Ammonia with Prevalence of Enterocytozoon hepatopenaei (EHP) in Shrimp (Litopenaeus vannamei) on Several Super-Intensive Ponds in East Java, Indonesia

Anord Charles Nkuba; Gunanti Mahasri; Nunuk Dyah Retno Lastuti; Adamu Ayubu Mwendolwa

doi:10.20473/jipk.v13i1.24430

Issue

Vol. 13 No. 1 (2021): JURNAL ILMIAH PERIKANAN DAN KELAUTAN

Issue Published : August 10, 2023

Date Log

Submitted

January 8, 2021

Accepted

March 23, 2021

Published

March 31, 2021

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.

1. Copyright of the article is transferred to the journal, by the knowledge of the author, whilst the moral right of the publication belongs to the author.

2. The legal formal aspect of journal publication accessibility refers to Creative Commons Atribusi-Non Commercial-Share alike (CC BY-NC-SA), (https://creativecommons.org/licenses/by-nc-sa/4.0/)

3. The articles published in the journal are open access and can be used for non-commercial purposes. Other than the aims mentioned above, the editorial board is not responsible for copyright violation

The manuscript authentic and copyright statement submission can be downloaded ON THIS FORM.

Fish Diseases Microplastics, Toxic, Pollution, and Heavy Metal

Correlation of Nitrite and Ammonia with Prevalence of Enterocytozoon hepatopenaei (EHP) in Shrimp (Litopenaeus vannamei) on Several Super-Intensive Ponds in East Java, Indonesia

https://doi.org/10.20473/jipk.v13i1.24430

Anord Charles Nkuba

Aqua-farms organization, Sinza C, Dar es Salaam, 22564. Tanzania

Gunanti Mahasri

Department of Fish Health Management and Aquaculture, Universitas Airlangga, Surabaya, East Java, 60115. Indonesia

Nunuk Dyah Retno Lastuti

Department of Parasitology, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, East Java, 60115. Indonesia

Adamu Ayubu Mwendolwa

Department of Biological Science, Mkwawa University College of Education, MUCE, 2513. Tanzania

Corresponding Author(s) : Anord Charles Nkuba

anordnkuba5@gmail.com

Jurnal Ilmiah Perikanan dan Kelautan, Vol. 13 No. 1 (2021): JURNAL ILMIAH PERIKANAN DAN KELAUTAN
Article Published : March 31, 2021

Abstract

Abstract

Enterocytozoon hepatopenaei (EHP), a microsporidia parasite known to hinder shrimp growth by infecting its hepatopancreas, is recently an emerging infection for Litopenaeus vannamei farms in Indonesia. The present study was conducted to investigate the relationship between nitrite and ammonia concentration with EHP infection in the super-intensive ponds. Shrimps and water samples were collected from six ponds in aquaculture super-intensive ponds with a stocking density of 5,000 shrimps in each pond which is located in Pasuruan, Lamongan, and Tuban Regency, Indonesia. The water sample was taken to the laboratory for the measurement of ammonia and nitrite. The samples of ammonia and nitrite were detected by spectrophotometer, and the PCR was used to detect 18S rRNA of EHP. The PCR analysis that showed two positive samples of L. vannamei, from the ponds with a high concentration of ammonia and nitrite, were infected by EHP. The statistical analysis showed a significant correlation between ammonia and nitrite with the prevalence of EHP infections, where the Pearson correlation (r) was 0.980 and 0.943, respectively. There was a high prevalence of EHP infection with the increase in nitrite concentration and ammonia in pond four and pond six. The concentration of over 1mg/l of ammonia and nitrite could influence EHP infection prevalence in the shrimp farms.

Highlight Research

Ammonia and nitrite concentration influence the prevalence of Enterocytozoon hepatopenaei infection in Super intensive ponds.
The estimated prevalence of Enterocytozoon hepatopenaei was 16.7%.
PCR amplification analysis was used to detect the 18S ss-rRNA of Enterocytozoon hepatopenaeiand 510-bp was obtained.

Keywords

HepatopancreasDNAPCRWater physicochemical parameters

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

Download Citation

References

Alfiansah, Y. R., Peters, S., Harder, J., Hassenrück, C., & Gärdes, A. (2020). Structure and co-occurrence patterns of bacterial communities associated with white feces disease outbreaks in Pacific white-leg shrimp Penaeus vannamei aquaculture. Scientific Reports, 10(1):1-14.
Anjaini, J., Agustin, I., & Bayu, I. (2018). Histopathological in gills, hepatopancreas and gut of white shrimp (Litopenaeus vannamei) infected white feces disease (WFD). Research Journal of Life Science, 5(3):183-194.
Aranguren, L. F., Han, J. E., & Tang, K. F. J. (2017). Enterocytozoon hepatopenaei (EHP) is a risk factor for acute hepatopancreatic necrosis disease (AHPND) and septic hepatopancreatic necrosis (SHPN) in the Pacific white shrimp Penaeus vannamei. Aquaculture, 471:37-42.
Ariadi, H., Fadjar, M., Mahmudi, M., & Supriatna. (2019). The relationships between water quality parameters and the growth rate of white shrimp (Litopenaeus vannamei) in intensive ponds. AACL Bioflux, 12(6): 2103-2116.
Aydin, A., Ercan, Ö., & TaşcioÇ§lu, S. (2005). A novel method for the spectrophotometric determination of nitrite in water. Talanta, 66(5): 1181-1186.
Biju, N., Sathiyaraj, G., Raj, M., Shanmugam, V., Baskaran, B., Govindan, U., Kumaresan, G., Kasthuriraju, K. K., & Chellamma, T. S. R. Y. (2016). High prevalence of enterocytozoon hepatopenaei in shrimps Penaeus monodon and Litopenaeus vannamei sampled from slow growth ponds in India. Diseases of Aquatic Organisms, 120(3):225-230.
BÅ“uf, G., & Payan, P. (2001). How should salinity influence fish growth? Comparative Biochemistry and Physiology - C Toxicology and Pharmacology, 130(4):411-423.
Caro, L.F. A., 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.
Duan, Y., Liu, Q., Wang, Y., Zhang, J., & Xiong, D. (2018). Impairment of the intestine barrier function in Litopenaeus vannamei exposed to ammonia and nitrite stress. Fish and Shellfish Immunology, 78: 279-288.
Fayer, R. (2004). Infectivity of microsporidia spores stored in seawater at environmental temperatures. The Journal of Parasitology, 90(3):654-657.
FAO. (1986). Shrimp culture and pond management.
Flegel, T. W. (1998, November). Advances in Shrimp Biotechnology. In Proceedings to the Special Session on Shrimp Biotechnology (pp. 147-149) 5th Asian Fisheries Forum. Chiengmai, Thailand.
Furtado, P. S., Fugimura, M. M. S., Monserrat, J. M., Souza, D. M., Garcia, L. de O., & Wasielesky, W. (2015). Acute effects of extreme pH and its influences on the survival and biochemical biomarkers of juvenile White Shrimp, Litopenaeus vannamei. Marine and Freshwater Behaviour and Physiology, 48(6):417-429.
Furtado, P. S., Campos, B. R., Serra, F. P., Klosterhoff, M., Romano, L. A., & Wasielesky, W. (2015). Effects of nitrate toxicity in the Pacific white shrimp, Litopenaeus vannamei, reared with biofloc technology (BFT). Aquaculture International, 23(1): 315-327.
Gomes, R. S., de Lima, J. P. V., Cavalli, R. O., & Correia, E. de S. (2016). Acute Toxicity of Ammonia and Nitrite to Painted River Prawn, Macrobrachium carcinus, Larvae. Journal of the World Aquaculture Society, 47(2): 239-247.
Gutiérrez-Salazar, G. J., Molina-Garza, Z. J., Hernández-Acosta, M., García-Salas, J. A., Mercado-Hernández, R., & Galaviz-Silva, L. (2011). Pathogens in Pacific white shrimp (Litopenaeus vannamei Boone, 1931) and their relationship with physicochemical parameters in three different culture systems in Tamaulipas, Mexico. Aquaculture, 321(1-2): 34-40.
Han, S., Wang, B., Wang, M., Liu, Q., Zhao, W., & Wang, L. (2017). Effects of ammonia and nitrite accumulation on the survival and growth performance of white shrimp Litopenaeus vannamei. Invertebrate Survival Journal, 14(1):221-232.
Hanggono, B., Lestari,Y. N., Fatmawati, Waluya, J. & Yuli, T.E. (2019). Deteksi cepat Enterocytozoon hepatopenaei (EHP) pada udang vaname (Penaeus vannamei). Jurnal Perekayasaan Budidaya Air Payau dan Laut, 14:35-40.
Jaroenlak, P., Sanguanrut, P., Williams, B. A. P., Stentiford, G. D., Flegel, T. W., Sritunyalucksana, K., & Itsathitphaisarn, O. (2016). A nested PCR assay to avoid false positive detection of the microsporidian Enterocytozoon hepatopenaei (EHP) in environmental samples in shrimp farms. PLOS One, 11(11): e0166320.
Karthikeyan, K., & Sudhakaran, R. (2020). Exploring the potentiality of Artemia salina to act as a reservoir for microsporidian Enterocytozoon hepatopenaei of penaeid shrimp. Biocatalysis and Agricultural Biotechnology, 25, 101607.
Karthikeyan, Kesavan, & Sudhakaran, R. (2019). Experimental horizontal transmission of Enterocytozoon hepatopenaei in post-larvae of whiteleg shrimp, Litopenaeus vannamei. Journal of Fish Diseases, 42(3):397-404.
Liao, I. C., & Chien, Y. (2011). The Pacific white shrimp, Litopenaeus vannamei, in Asia: The World's Most Widely Cultured Alien Crustacean. (pp. 489-519). In In the Wrong Place - Alien Marine Crustaceans: Distribution, Biology and Impacts. New York: Spinger.
Liang, Z., Liu, R., Zhao, D., Wang, L., Sun, M., Wang, M., & Song, L. (2016). Ammonia exposure induces oxidative stress, endoplasmic reticulum stress and apoptosis in hepatopancreas of pacific white shrimp (Litopenaeus vannamei). Fish and Shellfish Immunology, 54: 523–528.
Liu, C. H., & Chen, J. C. (2004). Effect of ammonia on the immune response of white shrimp Litopenaeus vannamei and its susceptibility to Vibrio alginolyticus. Fish and Shellfish Immunology, 16(3):321-334.
Liu, F., Li, S., Yu, Y., Sun, M., Xiang, J., & Li, F. (2020). Effects of ammonia stress on the hemocytes of the Pacific white shrimp Litopenaeus vannamei. Chemosphere, 239:124759.
Luna, G. (2017). Effects of pH, Alkalinity, and hardness on growth & Survivals of Vannamei shrimp in Aquaculture. Elsevier.
Mahasri, G., Hidayat, T., & Sudarno. (2019). Prevalence and intensity of ectoparasites in Pacific white shrimp (Litopenaeus vannamei) seeds from a pond and hatchery. IOP Conference Series: Earth and Environmental Science, 236(1).
Morris, A. W., & Riley, J. P. (1963). The determination of nitrate in sea water. Analytica Chimica Acta, 29(C), 272–279.
Nguyen, T. A. T., Nguyen, K. A. T., & Jolly, C. (2019). Is super-intensification the solution to shrimp production and export sustainability?. Sustainability, 11(19), 1-22.
Pamenang, G. D., Sulmartiwi, L., Mahasri, G., Rahayu, N. D., & Angwarmas, B. (2020). Inventory of ectoparasites in pacific white shrimp (Litopenaeus vannamei) that cultivated with high density. IOP Conference Series: Earth and Environmental Science, 441(1).
Rajendran, K. V., Shivam, S., Praveena, E. P., Rajan, J. S., Kumar, T.S., Avunje, S., Jagadeesan, V., Prasad Babu, S. V. A. N. V., Pande, A., Krishnan, A.N., Alavandi, S. V., & Vijayan, K. K. (2016). Emergence of Enterocytozoon hepatopenaei (EHP) in farmed Penaeus (Litopenaeus) vannamei in India. Aquaculture, 454(2016), 272-280.
Rubel, H., Woods, W., Pérez, D., Unnikrishnan, S., Felde, A. M. Z., Zielcke, S., Lidy, C. & Lanfer, C. (2019). A Strategic Approach to Sustainable Shrimp Production in Thailand: The Case for Improved Economics and Sustainability. Boston: Boston Consulting Group.
Sivakumar, S., Vimal, S., Abdul Majeed, S., Santhosh Kumar, S., Taju, G., Madan, N., Rajkumar, T., Thamizhvanan, S., Shamsudheen, K. V., Scaria, V., Sivasubbu, S., & Sahul Hameed, A. S. (2018). A new strain of white spot syndrome virus affecting Litopenaeus vannamei in Indian shrimp farms. Journal of Fish Diseases, 41(7):1129-1146.
SNI. (2000). Batas maksimum cemaran mikroba dan batas maksimum residu dalam bahan makanan asal hewan. Standar Nasional Indonesia No: 01-6366-2000. 16 p.
Sumini, S., & Kusdarwati, R. (2020). The discovery of Vibrio harveyi on Litopenaeus vannamei infected white feces disease in Situbondo, East Java. Jurnal Perikanan Universitas Gadjah Mada, 22(1), 9-18.
Tang, K. F. J., Aranguren, L. F., Piamsomboon, P., Han, J. E., Maskaykina, I. Y., & Schmidt, M. M. (2017). Detection of the microsporidian Enterocytozoon hepatopenaei (EHP) and Taura syndrome virus in Penaeus vannamei cultured in Venezuela. Aquaculture, 480: 17-21.
Tang, K. F. J., Han, J. E., Aranguren, L. F., White-Noble, B., Schmidt, M. M., Piamsomboon, P., Risdiana, E., & Hanggono, B. (2016). Dense populations of the microsporidian Enterocytozoon hepatopenaei (EHP) in feces of Penaeus vannamei exhibiting white feces syndrome and pathways of their transmission to healthy shrimp. Journal of Invertebrate Pathology, 140: 1-7.
Tang, K., Pantoja, C., Redman, R.M., Han. J.E., Tran L., & Lightner, D.V. (2015). Development of in situ hybridization and PCR assays for the detection of Enterocytozoon hepatopenaei (EHP), a microsporidian parasite infecting penaeid shrimp. Journal of Invertebrate Pathology, 130:37-41.
Tangprasittipap, A., Srisala, J., Chouwdee, S., SombooTang, K. F. J., Pantoja, C. R., Redman, R. M., Han, J. E., Tran, L. H., & Lightner, D. V. (2015). Development of in situ hybridization and PCR assays for the detection of Enterocytozoon hepatopenaei (EHP), a microsporidian parasite infecting penaeid shrimp. Journal of Invertebrate Pathology, 130:37-41.
Tangprasittipap, A., Srisala, J., Chouwdee, S., Somboon, M., Chuchird, N., Limsuwan, L., Srisuvan T., Flegel, T.W., Sritunyalucksana, K. (2013). The microsporidian Enterocytozoon hepatopenaei is not the cause of white feces syndrome in whiteleg shrimp Penaeus (Litopenaeus) vannamei. BMC Veterinary Research, 9:139.
Thitamadee, S., Prachumwat, A., Srisala, J., Jaroenlak, P., Salachan, P. V., Sritunyalucksana, K., Flegel, T. W., & Itsathitphaisarn, O. (2016). Review of current disease threats for cultivated penaeid shrimp in Asia. Aquaculture, 452: 69-87.
Tourtip, S., Wongtripop, S., Stentiford, G. D., Bateman, K. S., Sriurairatana, S., Chavadej, J., Sritunyalucksana, K., & Withyachumnarnkul, B. (2009). Enterocytozoon hepatopenaei sp. nov. (Microsporida: Enterocytozoonidae), a parasite of the black tiger shrimp Penaeus monodon (Decapoda: Penaeidae): Fine structure and phylogenetic relationships. Journal of Invertebrate Pathology, 102(1):21-29.
Waikhom, S. (2017). Effect of ammonia and nitrite toxicity on Pacific White Shrimp Litopenaeus vannamei in culture systems. MasterThesis, 15055, 87.
Xiao, J., Li, Q. Y., Tu, J. P., Chen, X. L., Chen, X. H., Liu, Q. Y., Liu, H., Zhou, X. Y., Zhao, Y. Z., & Wang, H. L. (2019). Stress response and tolerance mechanisms of ammonia exposure based on transcriptomics and metabolomics in Litopenaeus vannamei. Ecotoxicology and Environmental Safety, 180:491-500.
Zhao, M., Yao, D., Li, S., Zhang, Y., & Aweya, J. J. (2020). Effects of ammonia on shrimp physiology and immunity: a review. Reviews in Aquaculture, 12(4): 2194-2211.
Zitomer, F., & Lambert, J. L. (1962). Spectrophotometric determination of ammonia as trichloramine. Analytical Chemistry, 34(13):1738-1740.

References

Alfiansah, Y. R., Peters, S., Harder, J., Hassenrück, C., & Gärdes, A. (2020). Structure and co-occurrence patterns of bacterial communities associated with white feces disease outbreaks in Pacific white-leg shrimp Penaeus vannamei aquaculture. Scientific Reports, 10(1):1-14.

Anjaini, J., Agustin, I., & Bayu, I. (2018). Histopathological in gills, hepatopancreas and gut of white shrimp (Litopenaeus vannamei) infected white feces disease (WFD). Research Journal of Life Science, 5(3):183-194.

Aranguren, L. F., Han, J. E., & Tang, K. F. J. (2017). Enterocytozoon hepatopenaei (EHP) is a risk factor for acute hepatopancreatic necrosis disease (AHPND) and septic hepatopancreatic necrosis (SHPN) in the Pacific white shrimp Penaeus vannamei. Aquaculture, 471:37-42.

Ariadi, H., Fadjar, M., Mahmudi, M., & Supriatna. (2019). The relationships between water quality parameters and the growth rate of white shrimp (Litopenaeus vannamei) in intensive ponds. AACL Bioflux, 12(6): 2103-2116.

Aydin, A., Ercan, Ö., & TaşcioÇ§lu, S. (2005). A novel method for the spectrophotometric determination of nitrite in water. Talanta, 66(5): 1181-1186.

Biju, N., Sathiyaraj, G., Raj, M., Shanmugam, V., Baskaran, B., Govindan, U., Kumaresan, G., Kasthuriraju, K. K., & Chellamma, T. S. R. Y. (2016). High prevalence of enterocytozoon hepatopenaei in shrimps Penaeus monodon and Litopenaeus vannamei sampled from slow growth ponds in India. Diseases of Aquatic Organisms, 120(3):225-230.

BÅ“uf, G., & Payan, P. (2001). How should salinity influence fish growth? Comparative Biochemistry and Physiology - C Toxicology and Pharmacology, 130(4):411-423.

Caro, L.F. A., 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.

Duan, Y., Liu, Q., Wang, Y., Zhang, J., & Xiong, D. (2018). Impairment of the intestine barrier function in Litopenaeus vannamei exposed to ammonia and nitrite stress. Fish and Shellfish Immunology, 78: 279-288.

Fayer, R. (2004). Infectivity of microsporidia spores stored in seawater at environmental temperatures. The Journal of Parasitology, 90(3):654-657.

FAO. (1986). Shrimp culture and pond management.

Flegel, T. W. (1998, November). Advances in Shrimp Biotechnology. In Proceedings to the Special Session on Shrimp Biotechnology (pp. 147-149) 5th Asian Fisheries Forum. Chiengmai, Thailand.

Furtado, P. S., Fugimura, M. M. S., Monserrat, J. M., Souza, D. M., Garcia, L. de O., & Wasielesky, W. (2015). Acute effects of extreme pH and its influences on the survival and biochemical biomarkers of juvenile White Shrimp, Litopenaeus vannamei. Marine and Freshwater Behaviour and Physiology, 48(6):417-429.

Furtado, P. S., Campos, B. R., Serra, F. P., Klosterhoff, M., Romano, L. A., & Wasielesky, W. (2015). Effects of nitrate toxicity in the Pacific white shrimp, Litopenaeus vannamei, reared with biofloc technology (BFT). Aquaculture International, 23(1): 315-327.

Gomes, R. S., de Lima, J. P. V., Cavalli, R. O., & Correia, E. de S. (2016). Acute Toxicity of Ammonia and Nitrite to Painted River Prawn, Macrobrachium carcinus, Larvae. Journal of the World Aquaculture Society, 47(2): 239-247.

Gutiérrez-Salazar, G. J., Molina-Garza, Z. J., Hernández-Acosta, M., García-Salas, J. A., Mercado-Hernández, R., & Galaviz-Silva, L. (2011). Pathogens in Pacific white shrimp (Litopenaeus vannamei Boone, 1931) and their relationship with physicochemical parameters in three different culture systems in Tamaulipas, Mexico. Aquaculture, 321(1-2): 34-40.

Han, S., Wang, B., Wang, M., Liu, Q., Zhao, W., & Wang, L. (2017). Effects of ammonia and nitrite accumulation on the survival and growth performance of white shrimp Litopenaeus vannamei. Invertebrate Survival Journal, 14(1):221-232.

Hanggono, B., Lestari,Y. N., Fatmawati, Waluya, J. & Yuli, T.E. (2019). Deteksi cepat Enterocytozoon hepatopenaei (EHP) pada udang vaname (Penaeus vannamei). Jurnal Perekayasaan Budidaya Air Payau dan Laut, 14:35-40.

Jaroenlak, P., Sanguanrut, P., Williams, B. A. P., Stentiford, G. D., Flegel, T. W., Sritunyalucksana, K., & Itsathitphaisarn, O. (2016). A nested PCR assay to avoid false positive detection of the microsporidian Enterocytozoon hepatopenaei (EHP) in environmental samples in shrimp farms. PLOS One, 11(11): e0166320.

Karthikeyan, K., & Sudhakaran, R. (2020). Exploring the potentiality of Artemia salina to act as a reservoir for microsporidian Enterocytozoon hepatopenaei of penaeid shrimp. Biocatalysis and Agricultural Biotechnology, 25, 101607.

Karthikeyan, Kesavan, & Sudhakaran, R. (2019). Experimental horizontal transmission of Enterocytozoon hepatopenaei in post-larvae of whiteleg shrimp, Litopenaeus vannamei. Journal of Fish Diseases, 42(3):397-404.

Liao, I. C., & Chien, Y. (2011). The Pacific white shrimp, Litopenaeus vannamei, in Asia: The World's Most Widely Cultured Alien Crustacean. (pp. 489-519). In In the Wrong Place - Alien Marine Crustaceans: Distribution, Biology and Impacts. New York: Spinger.

Liang, Z., Liu, R., Zhao, D., Wang, L., Sun, M., Wang, M., & Song, L. (2016). Ammonia exposure induces oxidative stress, endoplasmic reticulum stress and apoptosis in hepatopancreas of pacific white shrimp (Litopenaeus vannamei). Fish and Shellfish Immunology, 54: 523–528.

Liu, C. H., & Chen, J. C. (2004). Effect of ammonia on the immune response of white shrimp Litopenaeus vannamei and its susceptibility to Vibrio alginolyticus. Fish and Shellfish Immunology, 16(3):321-334.

Liu, F., Li, S., Yu, Y., Sun, M., Xiang, J., & Li, F. (2020). Effects of ammonia stress on the hemocytes of the Pacific white shrimp Litopenaeus vannamei. Chemosphere, 239:124759.

Luna, G. (2017). Effects of pH, Alkalinity, and hardness on growth & Survivals of Vannamei shrimp in Aquaculture. Elsevier.

Mahasri, G., Hidayat, T., & Sudarno. (2019). Prevalence and intensity of ectoparasites in Pacific white shrimp (Litopenaeus vannamei) seeds from a pond and hatchery. IOP Conference Series: Earth and Environmental Science, 236(1).

Morris, A. W., & Riley, J. P. (1963). The determination of nitrate in sea water. Analytica Chimica Acta, 29(C), 272–279.

Nguyen, T. A. T., Nguyen, K. A. T., & Jolly, C. (2019). Is super-intensification the solution to shrimp production and export sustainability?. Sustainability, 11(19), 1-22.

Pamenang, G. D., Sulmartiwi, L., Mahasri, G., Rahayu, N. D., & Angwarmas, B. (2020). Inventory of ectoparasites in pacific white shrimp (Litopenaeus vannamei) that cultivated with high density. IOP Conference Series: Earth and Environmental Science, 441(1).

Rajendran, K. V., Shivam, S., Praveena, E. P., Rajan, J. S., Kumar, T.S., Avunje, S., Jagadeesan, V., Prasad Babu, S. V. A. N. V., Pande, A., Krishnan, A.N., Alavandi, S. V., & Vijayan, K. K. (2016). Emergence of Enterocytozoon hepatopenaei (EHP) in farmed Penaeus (Litopenaeus) vannamei in India. Aquaculture, 454(2016), 272-280.

Rubel, H., Woods, W., Pérez, D., Unnikrishnan, S., Felde, A. M. Z., Zielcke, S., Lidy, C. & Lanfer, C. (2019). A Strategic Approach to Sustainable Shrimp Production in Thailand: The Case for Improved Economics and Sustainability. Boston: Boston Consulting Group.

Sivakumar, S., Vimal, S., Abdul Majeed, S., Santhosh Kumar, S., Taju, G., Madan, N., Rajkumar, T., Thamizhvanan, S., Shamsudheen, K. V., Scaria, V., Sivasubbu, S., & Sahul Hameed, A. S. (2018). A new strain of white spot syndrome virus affecting Litopenaeus vannamei in Indian shrimp farms. Journal of Fish Diseases, 41(7):1129-1146.

SNI. (2000). Batas maksimum cemaran mikroba dan batas maksimum residu dalam bahan makanan asal hewan. Standar Nasional Indonesia No: 01-6366-2000. 16 p.

Sumini, S., & Kusdarwati, R. (2020). The discovery of Vibrio harveyi on Litopenaeus vannamei infected white feces disease in Situbondo, East Java. Jurnal Perikanan Universitas Gadjah Mada, 22(1), 9-18.

Tang, K. F. J., Aranguren, L. F., Piamsomboon, P., Han, J. E., Maskaykina, I. Y., & Schmidt, M. M. (2017). Detection of the microsporidian Enterocytozoon hepatopenaei (EHP) and Taura syndrome virus in Penaeus vannamei cultured in Venezuela. Aquaculture, 480: 17-21.

Tang, K. F. J., Han, J. E., Aranguren, L. F., White-Noble, B., Schmidt, M. M., Piamsomboon, P., Risdiana, E., & Hanggono, B. (2016). Dense populations of the microsporidian Enterocytozoon hepatopenaei (EHP) in feces of Penaeus vannamei exhibiting white feces syndrome and pathways of their transmission to healthy shrimp. Journal of Invertebrate Pathology, 140: 1-7.

Tang, K., Pantoja, C., Redman, R.M., Han. J.E., Tran L., & Lightner, D.V. (2015). Development of in situ hybridization and PCR assays for the detection of Enterocytozoon hepatopenaei (EHP), a microsporidian parasite infecting penaeid shrimp. Journal of Invertebrate Pathology, 130:37-41.

Tangprasittipap, A., Srisala, J., Chouwdee, S., SombooTang, K. F. J., Pantoja, C. R., Redman, R. M., Han, J. E., Tran, L. H., & Lightner, D. V. (2015). Development of in situ hybridization and PCR assays for the detection of Enterocytozoon hepatopenaei (EHP), a microsporidian parasite infecting penaeid shrimp. Journal of Invertebrate Pathology, 130:37-41.

Tangprasittipap, A., Srisala, J., Chouwdee, S., Somboon, M., Chuchird, N., Limsuwan, L., Srisuvan T., Flegel, T.W., Sritunyalucksana, K. (2013). The microsporidian Enterocytozoon hepatopenaei is not the cause of white feces syndrome in whiteleg shrimp Penaeus (Litopenaeus) vannamei. BMC Veterinary Research, 9:139.

Thitamadee, S., Prachumwat, A., Srisala, J., Jaroenlak, P., Salachan, P. V., Sritunyalucksana, K., Flegel, T. W., & Itsathitphaisarn, O. (2016). Review of current disease threats for cultivated penaeid shrimp in Asia. Aquaculture, 452: 69-87.

Tourtip, S., Wongtripop, S., Stentiford, G. D., Bateman, K. S., Sriurairatana, S., Chavadej, J., Sritunyalucksana, K., & Withyachumnarnkul, B. (2009). Enterocytozoon hepatopenaei sp. nov. (Microsporida: Enterocytozoonidae), a parasite of the black tiger shrimp Penaeus monodon (Decapoda: Penaeidae): Fine structure and phylogenetic relationships. Journal of Invertebrate Pathology, 102(1):21-29.

Waikhom, S. (2017). Effect of ammonia and nitrite toxicity on Pacific White Shrimp Litopenaeus vannamei in culture systems. MasterThesis, 15055, 87.

Xiao, J., Li, Q. Y., Tu, J. P., Chen, X. L., Chen, X. H., Liu, Q. Y., Liu, H., Zhou, X. Y., Zhao, Y. Z., & Wang, H. L. (2019). Stress response and tolerance mechanisms of ammonia exposure based on transcriptomics and metabolomics in Litopenaeus vannamei. Ecotoxicology and Environmental Safety, 180:491-500.

Zhao, M., Yao, D., Li, S., Zhang, Y., & Aweya, J. J. (2020). Effects of ammonia on shrimp physiology and immunity: a review. Reviews in Aquaculture, 12(4): 2194-2211.

Zitomer, F., & Lambert, J. L. (1962). Spectrophotometric determination of ammonia as trichloramine. Analytical Chemistry, 34(13):1738-1740.