Addition of Molasses Ameliorates Water and Bio-Floc Quality in Shrimp Pond Water

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Yustian Rovi Alfiansah
Jens Harder
Matthew James Slater
Astrid Gärdes


Suspended particulate matter, phytoplankton and bacteria can be exploited to form larger aggregates, so-called bio-flocs. These serve as feeds for cultured shrimps, govern inorganic nutrients and load bacteria including pathogens. The current study aimed to simulate aggregate formation from available particulate matter in shrimp pond water and investigate quality of aggregates as well as possible impact to the pond water. Molasses was added to cylindrical tanks containing shrimp pond waters, and the tanks were rolled for 48 h. Besides water quality (inorganic nutrients and physical parameters), the researchers investigated and separated bacterial community compositions (BCC) to free-living (FL) and bio-flocs/particle-attached (PA) bacteria via 16S rRNA amplicon sequencing, and measured macro-molecules contents (carbohydrates, lipids and proteins) in the bio-flocs. Molasses addition increased bacterial numbers in the bio-flocs to two-fold higher than the FL’s. Moreover, potential probiotics such as Halomonas, Psychrobacter, Mesonia and Chromohalobacter were detected associated to bio-flocs and dominated the BCC. In contrast, bio-flocs without molasses showed 4-fold less carbohydrates and harboured elevated potential pathogens such as Vibrio and Alteromonas. Results show that molasses (at C/N ratio 1:2) increases pH (to 8.2 ± 0.09 and 8.0 ± 0.04 after 24 h and 48 h, respectively) in pond water, improving beneficial biofloc formation. Molasses also increased carbohydrates and proteins in bio-flocs and maintained abundances of beneficial bacteria resulting in low inorganic nutrient concentrations. Thus, molasses is suitable for shrimp farming to improve rearing processes.

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Yustian Rovi Alfiansah, Jens Harder, Matthew James Slater, & Astrid Gärdes. (2022). Addition of Molasses Ameliorates Water and Bio-Floc Quality in Shrimp Pond Water. Tropical Life Sciences Research, 33(1), 121–141.
Original Article


Alfiansah Y R, Hassenrück C, Kunzmann A, Taslihan A, Harder J and Gärdes A. (2018). Bacterial abundance and community composition in pond water from shrimp aquaculture systems with different stocking densities. Frontiers in Microbiology 9: 2457.

Alfiansah Y R, Peters S, Harder J, Hassenrück C and Gärdes A. (2020). Structure and co-occurrence patterns of bacterial communities associated with white faeces disease outbreaks in Pacific white-leg shrimp Penaeus vannamei aquaculture. Scientific Reports 10: 11980.

Alldredge A L, Passow U and Haddock S H D. (1998). The characteristics and transparent exopolymer particle (TEP) content of marine snow formed from thecate dinoflagellates. Journal of Plankton Research 20(3): 393–406.

Alonso-Rodr?guez R and Paez-Osuna F. (2003). Nutrients, phytoplankton and harmful algal blooms in shrimp ponds: A review with special reference to the situation in the Gulf of California. Aquaculture 219: 317–336.

Avnimelech Y. (2015). Biofloc technology: A practical guide book, 3rd ed. Baton Rouge, LA: World Aquaculture Society. Bolger A M, Lohse M and Usadel B. (2014). Trimmomatic: A flexible trimmer for Illumina sequence data. Bioinformatics 30: 2114–2120.

Bradford M M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72: 248–254.

Cuzon G, Rosas C, Gaxiola G, Taboada G and Van Wormhoudt A. (2000). Utilization of carbohydrates by shrimp. In L E Cruz-Suárez, D Ricque-Marie, M TapiaSalazar, M A Olvera-Novoa and R Civera-Cerecedo (eds.). Avances en Nutrición Acuícola V. Memorias del V Simposium Internacional de Nutrición Acuícola. Meridan, Yucatan, 19–22 November 2000, 328–339.

De Coen W M and Janssen C R. (1997). The use of biomarkers in Daphnia magna toxicity testing. IV. cellular energy allocation: a new methodology to assess the energy budget of toxicant-stressed Daphnia populations. Journal of Aquatic Ecosystem Stress and Recovery 6: 43–55.

De Schryver P, Crab R, Defoirdt T, Boon N and Verstraete W. (2008). The basics of bioflocs technology: The added value for aquaculture. Aquaculture 277: 125–137.

Ducret A, Quardokus E M and Brun Y V. (2016). MicrobeJ, a tool for high throughput bacterial cell detection and quantitative analysis. Nature Microbiology 1: 1–7.

Engel A. (2009). Determination of marine gel particles. In O Wurl (ed.), Practical guidelines for the analysis of seawater. Boca Raton: CRC Press, 125–142.

Eren A M, Maignien L, Sul W J, Murphy L G, Grim S L, Morrison H G and Sogin M L. (2013). Oligotyping: Differentiating between closely related microbial taxa using 16S rRNA gene data. Methods in Ecology and Evolution 4(12): 1111–1119.

Funge-Smith S J and Briggs M R P. (1998). Nutrient budgets in intensive shrimp ponds: Implications for sustainability. Aquaculture 164: 117–133.

Gärdes A, Iversen M H, Grossart H P, Passow U and Ullrich M S. (2011). Diatomassociated bacteria are required for aggregation of Thalassiosira weissflogii. ISME Journal 5: 436–445.

Grossart H P. (2010). Ecological consequences of bacterioplankton lifestyles: Changes in concepts are needed. Environmental Microbiology Reports 2(6): 706–714.

Grossart H P, Schlingloff A, Bernhard M, Simon M and Brinkhoff T. (2004). Antagonistic activity of bacteria isolated from organic aggregates of the German Wadden Sea. FEMS Microbiology Ecology 47(3): 387–396.

Grossart H P and Simon M. (1998). Bacterial colonization and microbial decomposition of limnetic organic aggregates (lake snow). Aquatic Microbial Ecology 15: 127–140.

Hargreaves J A. (2013). Biofloc production systems for aquaculture. SRAC Publication No. 4503. United States Department of Agriculture. Available at:

Hasson K W, Wyld E M, Fan Y, Lingsweiller S W, Weaver S J, Cheng J and Varner P W. (2009). Streptococcosis in farmed Litopenaeus vannamei: A new emerging bacterial disease of penaeid shrimp. Diseases of Aquatic Organisms 86: 93–106.

Heenatigala P P M and Fernando M U L. (2016). Occurance of bacteria species responsible for vibriosis in shrimp pond culture systems in Sri Lanka and assesment of the suitable control measures. Sri Lanka Journal of Aquatic Sciences 21(1): 1–17.

Joshi J, Srisala J, Truong V H, Chen I T, Nuangsaeng B, Suthienkul O, Chu F, Flegel T W, Sritunyalucksana K and Thitamadee S. (2014). Variation in Vibrio parahaemolyticus isolates from a single Thai shrimp farm experiencing an outbreak of acute hepatopancreatic necrosis disease (AHPND). Aquaculture 428–429: 297–302.

Kepner R L and Pratt J R. (1994). Use of fluorochromes for direct enumeration of total bacteria in environmental samples: Past and present. Microbiological Reviews 58(4): 603–615.

Kimbrel J A, Ballor N, Wu Y W, David M M, Hazen T C, Simmons B A, Singer S W and Jansson J K. (2018). Microbial community structure and functional potential along a hypersaline gradient. Frontiers in Microbiology 9: 1492.

Kiørboe T. (2001). Formation and fate of marine snow: Small-scale processes with largescale implications. Scientia Marina 65(Suppl. 2): 57–71.

Kiørboe T, Tang K, Grossart H-P and Ploug H. (2003). Dynamics of microbial communities on marine snow aggregates: Colonization, growth, detachment, and grazing mortality of attached bacteria. Applied and Environmental Microbiology 69(6): 3036–3047.

Klindworth A, Pruesse E, Schweer T, Peplies J, Quast C, Horn M and Glöckner F O. (2013). Evaluation of general 16S ribosomal RNA gene PCR primers for classical and next-generation sequencing-based diversity studies. Nucleic Acids Research 41(1): e1.

Kramer A M, Lyons M M, Dobbs F C and Drake J M. (2013). Bacterial colonization and extinction on marine aggregates: Stochastic model of species presence and abundance. Ecology and Evolution 3(13): 4300–4309.

Kumar G R and Babu D E. (2015). The growth of microalgae in shrimp hatchery: Impact of environment on nutritional values. IOSR Journal of Biotechnology and Biochemistry 1: 89–96.

Li Y, Wu C, Zhou M, Wang E T, Zhang Z, Liu W, Ning J and Xie Z. (2017). Diversity of cultivable protease-producing bacteria in Laizhou Bay sediments, Bohai Sea, China. Frontiers in Microbiology 8: 1–10.

Lyons M M, Ward J E, Gaff H, Hicks R E, Drake J M and Dobbs F C. (2010). Theory of island biogeography on a microscopic scale: Organic aggregates as islands for aquatic pathogens. Aquatic Microbial Ecology 60(1): 1–13.

Lyons M M, Ward J E, Smolowitz R, Uhlinger K R and Gast R J. (2005). Lethal marine snow: Pathogen of bivalve mollusc concealed in marine aggregates. Limnology and Oceanography 50: 1983–1988.

Makino K, Oshima K, Kurokawa K, Yokoyama K, Uda T, Tagomori K, et al. (2003). Genome sequence of Vibrio parahaemolyticus: A pathogenic mechanism distinct from that of V cholerae. Lancet 361(9359): 743–749.

Mastan S A. (2015). Incidences of white feces syndrome (WFS) in farm-reared shrimp, Litopenaeus vannamei, Andhra Pradesh. Indo American Journal of Pharmaceutical Research 5: 9.

Miao S, Sun L, Bu H, Zhu J and Chen G. (2017). Effect of molasses addition at C:N ratio of 20:1 on the water quality and growth performance of giant freshwater prawn (Macrobrachium rosenbergii). Aquaculture International 25: 1409–1425.

Moriarty D J W. (1997). The role of microorganisms in aquaculture ponds. Aquaculture 151: 333–349. Moss S. (2002). Marine shrimp farming in the western hemisphere: Past problems, present solutions, and future visions. Reviews in Fisheries Science 10(3–4): 601–620.

Munro J, Oakey J, Bromage E and Owens L. (2003). Experimental bacteriophagemediated virulence in strains of Vibrio harveyi. Diseases of Aquatic Organism 54: 187–194.

Nercessian O, Noyes E, Kalyuzhnaya M G, Lidstrom M E and Chistoserdova L . (2005). Bacterial populations active in metabolism of C1 compounds in the sediment of Lake Washington, a freshwater lake. Applied and Environmental Microbiology 71: 6885–6899.

Oksanen J. (2015). Multivariate analysis of ecological communities in R: vegan tutorial. R Doc., 1–43. Available at:

_______. (2017). Vegan: Community ecology package. R package Version 2.4-3. 1–12.

Ortega-Retuerta E, Joux F, Jeffrey W H and Ghiglione J F. (2013). Spatial variability of particle-attached and free-living bacterial diversity in surface waters from the Mackenzie River to the Beaufort Sea (Canadian Arctic). Biogeosciences 10: 2747–2759.

Panjaitan P. (2010). Shrimp culture of Penaeus monodon with zero water exchange model (ZWEM) using molasses. Journal of Coastal Development 14(1): 35–44.

Pruesse E, Peplies J and Glöckner F O. (2012). SINA: Accurate high-throughput multiple sequence alignment of ribosomal RNA genes. Bioinformatics 28: 1823–1829.

Ramette A and Buttigieg P L. (2014). The R package OTU2ot for implementing the entropy decomposition of nucleotide variation in sequence data. Frontiers in Microbiology 5: 1–9.

Ray A J, Seaborn G, Leffler J W, Wilde S B, Lawson A and Browdy C L. (2010). Characterization of microbial communities in minimal-exchange, intensive aquaculture systems and the effects of suspended solids management. Aquaculture 310: 130–138.

Rieck A, Herlemann D P R, Jürgens K and Grossart H P. (2015). Particle-associated differ from free-living bacteria in surface waters of the Baltic sea. Frontiers in Microbiology 6: 1297.

Riemann L and Grossart H P. (2008). Elevated lytic phage production as a consequence of particle colonization by a marine Flavobacterium (Cellulophaga sp.). Microbial Ecology 56: 505–512.

Sangnoi Y, Chankaew S and O-Thong S. (2016). Indigenous Halomonas spp., the potential nitrifying bacteria for saline ammonium waste water treatment. Pakistan Journal of Biological Sciences 20(1): 52–58.

Shaari A L, Surif M, Latiff F A, Omar W M W and Ahmad M N. (2011). Monitoring of water quality and microalgae species composition of Penaeus Monodon ponds in Pulau Pinang, Malaysia. Tropical Life Sciences Research 22(1): 51–69.

Simon M, Grossart H, Schweitzer B and Ploug H. (2002). Microbial ecology of organic aggregates in aquatic ecosystems. Aquatic Microbial Ecology 28(2): 175–211.

Sombatjinda S, Boonapatcharoen N, Ruengjitchatchawalya M, Wantawin C, Withyachumnarnkul B and Techkarnjanaruk S. (2011). Dynamics of microbial communities in an earthen shrimp pond during the shrimp growing period. Environment and Natural Resources Research 1(1): 171–180.

Soto-Rodriguez S A, Gomez-Gil B, Lozano-Olvera R, Betancourt-Lozano M and Morales-Covarrubias M S. (2015). Field and experimental evidence of Vibrio parahaemolyticus as the causative agent of acute hepatopancreatic necrosis disease of cultured shrimp (Litopenaeus vannamei) in Northwestern Mexico. Applied and Environmental Microbiology 81(5): 1689–1699.

Suantika G, Aditiawati P, Astuti D I and Khotimah Z F. (2013). The use of indigenous probiotic Halomonas aquamarina and Shewanella algae for white shrimp (Litopenaeus vannamei Boone) hatchery productivity in zero water discharge system. Journal of Aquaculture Research & Development 4(5): 1000194.

United States Department of Agriculture/USDA (2011). Soil survey laboratory information manual. In R Burt (ed.). Soil survey investigations report no. 45, Version 2.0. U.S. Department of Agriculture, Natural Resources Conservation Service. 530 pp. United States Occupational Safety and Health

Administration/US OSHA. (2021). Safety and health topics, hydrogen sulfide. (accessed on 26 October 2021).

Utter D R, Mark Welch J L and Borisy G G. (2016). Individuality, stability, and variability of the plaque microbiome. Frontiers in Microbiology 7: 564.

Xiao J, Liu L, Ke Y, Li X, Liu Y, Pan Y, Yan S and Wang Y. (2017). Shrimp AHPNDcausing plasmids encoding the PirAB toxins as mediated by pirAB-Tn903 are prevalent in various Vibrio species. Scientific Reports 7: 42177.

Xiong J, Wang K, Wu J, Qiuqian L, Yang K, Qian Y and Zhang D. (2015). Changes in intestinal bacterial communities are closely associated with shrimp disease severity. Applied Microbiology and Biotechnology 99: 6911–6919.

Yaakob Z, Ali E, Zainal A, Mohamad M and Takriff M S. (2014). An overview: Biomolecules from microalgae for animal feed and aquaculture. Journal of Biological Research Thessaloniki 21: 6.

Zhang D, Wang X, Xiong J, Zhu J, Wang Y, Zhao Q, Chen H, Guo A, Wu J and Dai H. (2014). Bacterioplankton assemblages as biological indicators of shrimp health status. Ecological Indicators 38: 218–224.

Zhang J, Kobert K, Flouri T and Stamatakis A. (2014). PEAR: A fast and accurate Illumina Paired-End reAd mergeR. Bioinformatics 30(5): 614–620.

Zhang L, Mai K, Tan B, Ai Q, Qi C, Xu W, Zhang, W, Liufu Z, Wang X and Ma H. (2009). Effects of dietary administration of probiotic Halomonas sp. B12 on the intestinal microflora, immunological parameters, and midgut histological structure of shrimp, Fenneropenaeus chinensis. Journal of World Aquaculture Society 40(1): 58–66.

Zheng Y, Yu M, Liu Y, Su Y, Xu T, Yu M and Zhang X-H. (2016). Comparison of cultivable bacterial communities associated with Pacific white shrimp (Litopenaeus vannamei) larvae at different health statuses and growth stages. Aquaculture 451: 163–169.