Safety, Adherence, Enzymatic Activities, and Application Effects of Oral Probiotic Candidates for Shortfin Eel (Anguilla bicolor bicolor)

Andita Ratih Dewanti; Anggi Octari Putri; Indah Istiqomah Istiqomah; Alim Isnansetyo

doi:10.20473/jipk.v14i2.34315

Issue

Vol. 14 No. 2 (2022): JURNAL ILMIAH PERIKANAN DAN KELAUTAN

Issue Published : August 30, 2022

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Submitted

March 16, 2022

Accepted

April 20, 2022

Published

August 30, 2022

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Fish Health Management

Safety, Adherence, Enzymatic Activities, and Application Effects of Oral Probiotic Candidates for Shortfin Eel (Anguilla bicolor bicolor)

https://doi.org/10.20473/jipk.v14i2.34315

Andita Ratih Dewanti

Department of Fisheries, Faculty of Agriculture, Universitas Gadjah Mada, Yogyakarta, 55281. Indonesia

Anggi Octari Putri

Department of Fisheries, Faculty of Agriculture, Universitas Gadjah Mada, Yogyakarta, 55281. Indonesia

Indah Istiqomah Istiqomah

Department of Fisheries, Faculty of Agriculture, Universitas Gadjah Mada, Yogyakarta, 55281. Indonesia

https://orcid.org/0000-0002-1053-9048

Alim Isnansetyo

Department of Fisheries, Faculty of Agriculture, Universitas Gadjah Mada, Yogyakarta, 55281. Indonesia

https://orcid.org/0000-0002-3219-1185

Corresponding Author(s) : Indah Istiqomah Istiqomah

indah_ist@ugm.ac.id

Jurnal Ilmiah Perikanan dan Kelautan, Vol. 14 No. 2 (2022): JURNAL ILMIAH PERIKANAN DAN KELAUTAN
Article Published : August 30, 2022

Abstract

Highlight Research

The shortfin eel elver bicolor bicolor was tested for the safety of Enterobacter sp. JC05, Lactococcus sp. JAL37, and Bacillus sp. PCP1

The ability of bacterial strains to adhere to epithelial cells of shortfin eel epithelial cells was demonstrated

The bacterial strains' proteolytic, cellulolytic, and lipolytic activities were detected

Oral administration of the bacterial cocktail lowered overall viable bacterial count but did not affect the shortfin eel's intestinal histological characteristics

Abstract

Aquaculture of the shortfin eel (Anguilla bicolor bicolor) has been plagued by low survival and growth due to the low tolerance to water quality and feed. The microbiota and shape of the fish intestinal tract influence the immune and digestive systems. The use of bacterial probiotics is fascinating to enhance the digestion system. This study aimed to characterize bacterial probiotic candidates' safety and potential probiotic features for shortfin eel (A. bicolor bicolor) aquaculture. The safety, adherence, and enzymatic activity of three bacterial strains (Bacillus sp. PCP1, Lactococcus sp. JAL 37, and Enterobacter sp. JC05) were investigated. An oral application test was performed on shortfin eel (n=880, 15 g) every four days with 0, 3x10³, 3x10⁵, and 3x10⁷ CFU/g diet dosages in quadruplicates for two months. At the end of the experiment, total cultivable bacteria and intestinal morphology were assessed. Based on the hemolytic test and intraperitoneal injection, the bacterial strains were considered harmless. In an in vitro investigation, the bacteria attached to shortfin eel intestinal epithelial cells. An agar diffusion method validated the proteolytic, lipolytic, and cellulolytic activities. In vivo feeding tests with the bacterial cocktail lowered the total viable bacteria in the gut while preserving the gut histology. Bacterial strains of the present study are potential probiotic candidates for shortfin (A. bicolor bicolor) aquaculture.

Keywords

Intestine Epithelial Cells Probiotic Eel Viable Bacteria

Dewanti, A. R., Putri, A. O., Istiqomah, I. I., & Isnansetyo, A. (2022). Safety, Adherence, Enzymatic Activities, and Application Effects of Oral Probiotic Candidates for Shortfin Eel (Anguilla bicolor bicolor). Jurnal Ilmiah Perikanan Dan Kelautan, 14(2), 203–213. https://doi.org/10.20473/jipk.v14i2.34315

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References

Aisyah, I., Istiqomah, I., & Isnansetyo, A. (2020). Effect of dietary probiotics SEAL on growth performance of red tilapia (Oreochromis sp.) nilasa strain. IOP Conference Series: Earth Environmental Science, 584:012042.
Apún-Molina, J. P., Santamaría-Miranda, A., Luna-González, A., Ibarra-Gámez, J. C., Medina-Alcantar, V., & Racotta, I. (2015). Growth and metabolic responses of whiteleg shrimp Litopenaeus vannamei and Nile tilapia Oreochromis niloticus in polyculture fed with potential probiotic microorganisms on different schedules. Latin American Journal of Aquatic Research, 43(3):435-445.
Atitus, I. (2018). Isolasi dan identifikasi bakteri selulolitik dari beberapa jenis ikan laut. Thesis. Yogyakarta: Universitas Gadjah Mada.
Banerjee, G., & Ray, A. K. (2017). The advancement of probiotics research and its application in fish farming industries. Research in veterinary science, 115:66-77.
De, B. C., Meena, D. K., Behera, B. K., Das, P., Mohapatra, P. K. D., & Sharma, A. P. (2014). Probiotics in fish and shellfish culture: immunomodulatory and ecophysiological responses. Fish physiology and biochemistry, 40(3):921-971.
Elsabagh, M., Mohamed, R., Moustafa, E. M., Hamza, A., Farrag, F., Decamp, O., Dawood, M. A. O., & Eltholth, M. (2018). Assessing the impact of Bacillus strains mixture probiotic on water quality, growth performance, blood profile and intestinal morphology of Nile tilapia, Oreochromis niloticus. Aquaculture Nutrition, 24(6):1613-1622.
El-Saadony, M. T., Alagawany M., Patra, A. K., Kar, I., Tiwari, R., Dawood, M. A. O., Dhama, K., & Abdel-Latif, H. M. R. (2021). The functionality of probiotics in aquaculture: An overview. Fish & Shellfish Immunology, 117:36-52.
Faturrahman, F., Khair, W. H., & Andini, A. S. (2021). Heterotrophic bacteria from the digestive tract of rice field eel (Monopterus albus) and its potency as probiotic. Journal of Physics: Conference Series, 1779(1):1-7.
Febrianta, H., & Rawendra, R. D. S. (2019). Nutrition evaluation of Indonesian shortfin eel (Anguilla bicolor) meat for functional food development. Journal of Physics: Conference Series, 1363:012010.
Ghosh, K., Mukherjee, A., Dutta, D., Banerjee, S., Breines, E. M., Hareide, E., & Ringo, E. (2021). Endosymbiotic pathogen-inhibitory gut bacteria in three indian major carps under polyculture system: a step toward making a probiotics consortium. Aquaculture and Fisheries, 6(2):192-204.
Grumezescu, A. M., & Holban, A. M. (2018). Therapeutic, probiotic, and unconventional foods (1st ed.). Cambridge: Academic Press.
Hani, Y. M. I., Marchand, A., Turies, C., Kerambrun, E., Palluel, O., Bado-Nilles, A., Beaudouin, R., Porcher, J. M., Geffard, A., & Dedourge-Geffard, O. (2018). Digestive enzymes and gut morphometric parameters of threespine stickleback (Gasterosteus aculeatus): Influence of body size and temperature. Plos One, 13(4):1-21.
Hikmawati, F., Susilowati, A., & Setyaningsih, R. (2019). Detection of the number and pathogenicity of Vibrio spp. on green mussels (Perna viridis) in the tourist area of Yogyakarta. Prosiding Seminar Nasional Masyarakat Biodiversitas Indonesia, 5(2):334-339.
Iorizzo, M., Albanese, G., Letizia, F., Testa, B., Tremonte, P., Vergalito, F., Lombardi, S. J., Succi, M., Coppola, R., & Sorrentino, E. (2022). Probiotic potentiality from versatile Lactiplantibacillus plantarum strains as resource to enhance freshwater fish health. Microorganisms, 10(2):463.
Istiqomah, I., Isnansetyo, A., Atitus, I., & Rohman, A. F. (2019). Isolation of cellulolytic bacterium Staphylococcus sp. JC20 from the intestine of octopus (Octopus sp.) for fish probiotic candidate. Jurnal Perikanan Universitas Gadjah Mada, 21(2):93-98.
Istiqomah, I., Kawato, Y., Mahmoud, M. M., Okuda, J., Yasuike, M., Nakamura, Y., Fujiwara, A., Sahiro, K., Sadakane, M., & Nakai, T. (2015). Fimbriae expression by Edwardsiella tarda in high-salt culture conditions. Fish Pathology, 50(4):207-212.
Kavitha, M., Raja, M., & Perumal, P. (2018). Evaluation of probiotic potential of Bacillus spp. isolated from the digestive tract of freshwater fish Labeo calbasu (Hamilton, 1822). Aquaculture Reports, 11:59-69.
Lazado, C. C., & Caipang, C. M. A. (2014). Mucosal immunity and probiotics in fish. Fish & Shellfish Immunology, 39(1):78-89.
Lazado, C. C., Caipang, C. M. A., & Estante, E. G. (2015). Prospects of host-associated microorganisms in fish and penaeids as probiotics with immunomodulatory functions. Fish & Shellfish Immunology, 45(1):2-12.
Lee, J. S., Cheng, H., Damte, D., Lee, S. J., Kim, J. C., Rhee, M. H., Suh, J. W., & Park, S. C. (2013). Effects of dietary supplementation of Lactobacillus pentosus PL11 on the growth performance, immune and antioxidant systems of Japanese eel Anguilla japonica challenged with Edwardsiella tarda. Fish & Shellfish Immunology, 34(3):756-761.
Lee, S., Katya, K., Hamidoghli, A., Hong., J., Kim, D. J., & Bai, S. C. (2018). Synergistic effects of dietary supplementation of Bacillus subtilis WB60 and mannanoligosaccharide (MOS) on growth performance, immunity and disease resistance in Japanese eel, Anguilla japonica. Fish & Shellfish Immunology, 83:283-291.
Lee, S., Katya, K., Park, Y., Won, S., Seong, M., Hamidoghli, A., & Bai, S. C. (2017). Comparative evaluation of dietary probiotics Bacillus subtilis WB60 and Lactobacillus plantarum KCTC3928 on the growth performance, immunological parameters, gut morphology, and disease resistance in Japanese eel, Anguilla japonica. Fish & Shellfish Immunology, 61:201-210.
Li, X., Fields, F. R., Ho, M., Marshall-Hudson, A., Gross, R., Casser, M. E., & Naito, M. (2021). Safety assessment of Streptococcus salivarius DB-B5 as a probiotic candidate for oral health. Food and Chemical Toxicology, 153:112277.
Melo"Bolívar, J. F., Pardo, R. Y. R., Hume, M. E., & Diaz, L. M. V. (2021). Multistrain probiotics use in main commercially cultured freshwater fish: A systematic review of evidence. Reviews in Aquaculture, 13(4):1758-1780.
Muchlisin, Z. A., Sofyan, M., Dewiyanti, I., Nur, F. M., Batubara, A. S., Fadli, N., Muhammadar, A. A., Efizon, D., Fauzi, M., & Siti-Azizah, M. N. (2020). Data of feed formulation for Indonesian short-fin eel, Anguilla bicolor McClelland, 1844 Elver. Data in Brief, 30:105581.
Nandi, A., Dan, S. K., Banerjee, G., Ghosh, P., Ghosh, K., Ringí¸, E., & Ray, A. K. (2017). Probiotic potential of autochthonous bacteria isolated from the gastrointestinal tract of four freshwater teleosts. Probiotics and Antimicrobial Proteins, 9(1):12-21.
Newaj"Fyzul, A., & Austin, B. (2015). Probiotics, immunostimulants, plant products and oral vaccines, and their role as feed supplements in the control of bacterial fish diseases. Journal of Fish Diseases, 38(11):937-955.
Ouwehand, A. C. (2017). A review of dose-responses of probiotics in human studies. Beneficial Microbes, 8(2):143-151.
Rohman, A. F., Atitus, I. N., Heraswati, D. D., Istiqomah, I., & Isnansetyo, A. (2021). Isolation of Aeromonas sobria JC18 from milkfish (Chanos chanos) intestine with proteolytic and cellulolytic activities for fish probiotic. IOP Conference Series: Earth and Environmental Science, 919:012055.
Sarojini, K., Ajitha, S. S., Ramasubburayan, R., Palavesam, A., & Immanuel G. (2020). Studies on the adhesion, aggregative properties and the probiotic efficiency of a potent bacteriocin-producing shrimp gut isolate Bacillus subtilis subsp. inaquosorum V1 against Carassius auratus. Aquaculture Internasional, 28:1639-1656.
Shi, Y., Ma, D. Y., & Zhai, S. W. S. W. (2020). Revealing the difference of intestinal microbiota composition of cultured European eels (Anguilla anguilla) with different growth rates. Israeli Journal of Aquaculture, 72:1-12.
Slizewska, K., Wojcik, A. C., & Nowak, A. (2020). Probiotic properties of new Lactobacillus strains intended to be used as feed additives for monogastric animals. Probiotics and Antimicrobial Proteins, 13:146-162.
Soeprijanto, A., Guntur, G., & Fakhri., M. (2018). Application of probiotic and fermented feed in the nursery of Anguilla bicolor. Jurnal Perikanan Universitas Gadjah Mada, 20(1):19-22.
Soleha, S. & Retnaningrum, E. (2020). Screening and molecular identification of lipolytic bacteria from spent bleaching earth. Journal of Biological Diversity, 21(9):4155-4161.
Stepanauskas, R. (2013). Single-cell genomics dives into the diversity of marine microbes. San Diego: iCommunity Newsletter.
Suryaningsih, W., Maulana, R., Istiqomah, I., & Isnansetyo, A. (2021). In vitro adhesion of Bacillus sp. and Enterobacter sp. probiotics on intestinal epithelial cells of red tilapia (Oreochromis sp.) and the application effects on the fish growth and survival rate. IOP Conf. Series: Earth and Environmental Science, 919:012056.
Triyatmo, B., & Isnansetyo, A. (2020). Effects of probiotic dosage on water quality, total count of Aeromonas spp. and Pseudomonas spp. in Eel Anguilla bicolor cultivation. E3S Web of Conferences, 147(01008):1-4.
Taukhid, T., Prasetiyo, D., Andriyanto, S., Ahyani, N., Azril, M., & Amriana, A. (2021). Identification of micro-organisms (parasites and bacteria) which are potential as pathogenic agent in glass eel of Anguilla spp. at the eel capture location, Sukabumi, West Java. Journal of Aquaculture Science, 6(11S):170-179.
Wahjuningrum, D., Hidayat, A. M., & Budiardi, T. (2018). Characterization of pathogenic bacteria in eel Anguilla bicolor bicolor. Jurnal Akuakultur Indonesia, 17(1):94-103.
Widiantoro, W. (2020). Derajat kelangsungan hidup dan kesehatan ikan sidat (Anguilla bicolor) pada dua wilayah (tempat) pembesaran yang berbeda. Jurnal Aquafish Saintek, 1(1):35-38.
Yilmaz, S., Yilmaz, E., Dawood, M. A., Ringí¸, E., Ahmadifar, E., & Abdel-Latif, H. M. (2022). Probiotics, prebiotics, and synbiotics used to control vibriosis in fish: A review. Aquaculture, 547:737514.
Yin, H., Ye, P., Lei, Q., Cheng, Y., Yu, H., Du, J., Pan, H., & Cao, Z. (2020). In vitro probiotic properties of Pediococcus pentosaceus L1 and its effects on enterotoxigenic Escherichia coli-induced inflammatory responses in porcine intestinal epithelial cells. Microbial Pathogenesis, 144:104163.
Zhao, D., Wu, S., Feng, W., Jakovlic, I., Tran, N. T., Xiong, F. (2020). Adhesion and colonization properties of potentially probiotic Bacillus paralicheniformis strain FA6 isolated from grass carp intestine. Fisheries Science, 86:153-161.

References

Aisyah, I., Istiqomah, I., & Isnansetyo, A. (2020). Effect of dietary probiotics SEAL on growth performance of red tilapia (Oreochromis sp.) nilasa strain. IOP Conference Series: Earth Environmental Science, 584:012042.

Apún-Molina, J. P., Santamaría-Miranda, A., Luna-González, A., Ibarra-Gámez, J. C., Medina-Alcantar, V., & Racotta, I. (2015). Growth and metabolic responses of whiteleg shrimp Litopenaeus vannamei and Nile tilapia Oreochromis niloticus in polyculture fed with potential probiotic microorganisms on different schedules. Latin American Journal of Aquatic Research, 43(3):435-445.

Atitus, I. (2018). Isolasi dan identifikasi bakteri selulolitik dari beberapa jenis ikan laut. Thesis. Yogyakarta: Universitas Gadjah Mada.

Banerjee, G., & Ray, A. K. (2017). The advancement of probiotics research and its application in fish farming industries. Research in veterinary science, 115:66-77.

De, B. C., Meena, D. K., Behera, B. K., Das, P., Mohapatra, P. K. D., & Sharma, A. P. (2014). Probiotics in fish and shellfish culture: immunomodulatory and ecophysiological responses. Fish physiology and biochemistry, 40(3):921-971.

Elsabagh, M., Mohamed, R., Moustafa, E. M., Hamza, A., Farrag, F., Decamp, O., Dawood, M. A. O., & Eltholth, M. (2018). Assessing the impact of Bacillus strains mixture probiotic on water quality, growth performance, blood profile and intestinal morphology of Nile tilapia, Oreochromis niloticus. Aquaculture Nutrition, 24(6):1613-1622.

El-Saadony, M. T., Alagawany M., Patra, A. K., Kar, I., Tiwari, R., Dawood, M. A. O., Dhama, K., & Abdel-Latif, H. M. R. (2021). The functionality of probiotics in aquaculture: An overview. Fish & Shellfish Immunology, 117:36-52.

Faturrahman, F., Khair, W. H., & Andini, A. S. (2021). Heterotrophic bacteria from the digestive tract of rice field eel (Monopterus albus) and its potency as probiotic. Journal of Physics: Conference Series, 1779(1):1-7.

Febrianta, H., & Rawendra, R. D. S. (2019). Nutrition evaluation of Indonesian shortfin eel (Anguilla bicolor) meat for functional food development. Journal of Physics: Conference Series, 1363:012010.

Ghosh, K., Mukherjee, A., Dutta, D., Banerjee, S., Breines, E. M., Hareide, E., & Ringo, E. (2021). Endosymbiotic pathogen-inhibitory gut bacteria in three indian major carps under polyculture system: a step toward making a probiotics consortium. Aquaculture and Fisheries, 6(2):192-204.

Grumezescu, A. M., & Holban, A. M. (2018). Therapeutic, probiotic, and unconventional foods (1st ed.). Cambridge: Academic Press.

Hani, Y. M. I., Marchand, A., Turies, C., Kerambrun, E., Palluel, O., Bado-Nilles, A., Beaudouin, R., Porcher, J. M., Geffard, A., & Dedourge-Geffard, O. (2018). Digestive enzymes and gut morphometric parameters of threespine stickleback (Gasterosteus aculeatus): Influence of body size and temperature. Plos One, 13(4):1-21.

Hikmawati, F., Susilowati, A., & Setyaningsih, R. (2019). Detection of the number and pathogenicity of Vibrio spp. on green mussels (Perna viridis) in the tourist area of Yogyakarta. Prosiding Seminar Nasional Masyarakat Biodiversitas Indonesia, 5(2):334-339.

Iorizzo, M., Albanese, G., Letizia, F., Testa, B., Tremonte, P., Vergalito, F., Lombardi, S. J., Succi, M., Coppola, R., & Sorrentino, E. (2022). Probiotic potentiality from versatile Lactiplantibacillus plantarum strains as resource to enhance freshwater fish health. Microorganisms, 10(2):463.

Istiqomah, I., Isnansetyo, A., Atitus, I., & Rohman, A. F. (2019). Isolation of cellulolytic bacterium Staphylococcus sp. JC20 from the intestine of octopus (Octopus sp.) for fish probiotic candidate. Jurnal Perikanan Universitas Gadjah Mada, 21(2):93-98.

Istiqomah, I., Kawato, Y., Mahmoud, M. M., Okuda, J., Yasuike, M., Nakamura, Y., Fujiwara, A., Sahiro, K., Sadakane, M., & Nakai, T. (2015). Fimbriae expression by Edwardsiella tarda in high-salt culture conditions. Fish Pathology, 50(4):207-212.

Kavitha, M., Raja, M., & Perumal, P. (2018). Evaluation of probiotic potential of Bacillus spp. isolated from the digestive tract of freshwater fish Labeo calbasu (Hamilton, 1822). Aquaculture Reports, 11:59-69.

Lazado, C. C., & Caipang, C. M. A. (2014). Mucosal immunity and probiotics in fish. Fish & Shellfish Immunology, 39(1):78-89.

Lazado, C. C., Caipang, C. M. A., & Estante, E. G. (2015). Prospects of host-associated microorganisms in fish and penaeids as probiotics with immunomodulatory functions. Fish & Shellfish Immunology, 45(1):2-12.

Lee, J. S., Cheng, H., Damte, D., Lee, S. J., Kim, J. C., Rhee, M. H., Suh, J. W., & Park, S. C. (2013). Effects of dietary supplementation of Lactobacillus pentosus PL11 on the growth performance, immune and antioxidant systems of Japanese eel Anguilla japonica challenged with Edwardsiella tarda. Fish & Shellfish Immunology, 34(3):756-761.

Lee, S., Katya, K., Hamidoghli, A., Hong., J., Kim, D. J., & Bai, S. C. (2018). Synergistic effects of dietary supplementation of Bacillus subtilis WB60 and mannanoligosaccharide (MOS) on growth performance, immunity and disease resistance in Japanese eel, Anguilla japonica. Fish & Shellfish Immunology, 83:283-291.

Lee, S., Katya, K., Park, Y., Won, S., Seong, M., Hamidoghli, A., & Bai, S. C. (2017). Comparative evaluation of dietary probiotics Bacillus subtilis WB60 and Lactobacillus plantarum KCTC3928 on the growth performance, immunological parameters, gut morphology, and disease resistance in Japanese eel, Anguilla japonica. Fish & Shellfish Immunology, 61:201-210.

Li, X., Fields, F. R., Ho, M., Marshall-Hudson, A., Gross, R., Casser, M. E., & Naito, M. (2021). Safety assessment of Streptococcus salivarius DB-B5 as a probiotic candidate for oral health. Food and Chemical Toxicology, 153:112277.

Melo"Bolívar, J. F., Pardo, R. Y. R., Hume, M. E., & Diaz, L. M. V. (2021). Multistrain probiotics use in main commercially cultured freshwater fish: A systematic review of evidence. Reviews in Aquaculture, 13(4):1758-1780.

Muchlisin, Z. A., Sofyan, M., Dewiyanti, I., Nur, F. M., Batubara, A. S., Fadli, N., Muhammadar, A. A., Efizon, D., Fauzi, M., & Siti-Azizah, M. N. (2020). Data of feed formulation for Indonesian short-fin eel, Anguilla bicolor McClelland, 1844 Elver. Data in Brief, 30:105581.

Nandi, A., Dan, S. K., Banerjee, G., Ghosh, P., Ghosh, K., Ringí¸, E., & Ray, A. K. (2017). Probiotic potential of autochthonous bacteria isolated from the gastrointestinal tract of four freshwater teleosts. Probiotics and Antimicrobial Proteins, 9(1):12-21.

Newaj"Fyzul, A., & Austin, B. (2015). Probiotics, immunostimulants, plant products and oral vaccines, and their role as feed supplements in the control of bacterial fish diseases. Journal of Fish Diseases, 38(11):937-955.

Ouwehand, A. C. (2017). A review of dose-responses of probiotics in human studies. Beneficial Microbes, 8(2):143-151.

Rohman, A. F., Atitus, I. N., Heraswati, D. D., Istiqomah, I., & Isnansetyo, A. (2021). Isolation of Aeromonas sobria JC18 from milkfish (Chanos chanos) intestine with proteolytic and cellulolytic activities for fish probiotic. IOP Conference Series: Earth and Environmental Science, 919:012055.

Sarojini, K., Ajitha, S. S., Ramasubburayan, R., Palavesam, A., & Immanuel G. (2020). Studies on the adhesion, aggregative properties and the probiotic efficiency of a potent bacteriocin-producing shrimp gut isolate Bacillus subtilis subsp. inaquosorum V1 against Carassius auratus. Aquaculture Internasional, 28:1639-1656.

Shi, Y., Ma, D. Y., & Zhai, S. W. S. W. (2020). Revealing the difference of intestinal microbiota composition of cultured European eels (Anguilla anguilla) with different growth rates. Israeli Journal of Aquaculture, 72:1-12.

Slizewska, K., Wojcik, A. C., & Nowak, A. (2020). Probiotic properties of new Lactobacillus strains intended to be used as feed additives for monogastric animals. Probiotics and Antimicrobial Proteins, 13:146-162.

Soeprijanto, A., Guntur, G., & Fakhri., M. (2018). Application of probiotic and fermented feed in the nursery of Anguilla bicolor. Jurnal Perikanan Universitas Gadjah Mada, 20(1):19-22.

Soleha, S. & Retnaningrum, E. (2020). Screening and molecular identification of lipolytic bacteria from spent bleaching earth. Journal of Biological Diversity, 21(9):4155-4161.

Stepanauskas, R. (2013). Single-cell genomics dives into the diversity of marine microbes. San Diego: iCommunity Newsletter.

Suryaningsih, W., Maulana, R., Istiqomah, I., & Isnansetyo, A. (2021). In vitro adhesion of Bacillus sp. and Enterobacter sp. probiotics on intestinal epithelial cells of red tilapia (Oreochromis sp.) and the application effects on the fish growth and survival rate. IOP Conf. Series: Earth and Environmental Science, 919:012056.

Triyatmo, B., & Isnansetyo, A. (2020). Effects of probiotic dosage on water quality, total count of Aeromonas spp. and Pseudomonas spp. in Eel Anguilla bicolor cultivation. E3S Web of Conferences, 147(01008):1-4.

Taukhid, T., Prasetiyo, D., Andriyanto, S., Ahyani, N., Azril, M., & Amriana, A. (2021). Identification of micro-organisms (parasites and bacteria) which are potential as pathogenic agent in glass eel of Anguilla spp. at the eel capture location, Sukabumi, West Java. Journal of Aquaculture Science, 6(11S):170-179.

Wahjuningrum, D., Hidayat, A. M., & Budiardi, T. (2018). Characterization of pathogenic bacteria in eel Anguilla bicolor bicolor. Jurnal Akuakultur Indonesia, 17(1):94-103.

Widiantoro, W. (2020). Derajat kelangsungan hidup dan kesehatan ikan sidat (Anguilla bicolor) pada dua wilayah (tempat) pembesaran yang berbeda. Jurnal Aquafish Saintek, 1(1):35-38.

Yilmaz, S., Yilmaz, E., Dawood, M. A., Ringí¸, E., Ahmadifar, E., & Abdel-Latif, H. M. (2022). Probiotics, prebiotics, and synbiotics used to control vibriosis in fish: A review. Aquaculture, 547:737514.

Yin, H., Ye, P., Lei, Q., Cheng, Y., Yu, H., Du, J., Pan, H., & Cao, Z. (2020). In vitro probiotic properties of Pediococcus pentosaceus L1 and its effects on enterotoxigenic Escherichia coli-induced inflammatory responses in porcine intestinal epithelial cells. Microbial Pathogenesis, 144:104163.

Zhao, D., Wu, S., Feng, W., Jakovlic, I., Tran, N. T., Xiong, F. (2020). Adhesion and colonization properties of potentially probiotic Bacillus paralicheniformis strain FA6 isolated from grass carp intestine. Fisheries Science, 86:153-161.