Welcome To ABAWARE


Advanced Biotechnology for Intensive-Freshwater Aquaculture Wastewater Reuse – part of the European Commission’s  Water Joint Programming Initiative 2016 Joint Call which aims to support research on the sustainable management of water resources in agriculture, forestry and freshwater aquaculture systems.

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Advanced Biotechnologies for Reuse of Waste Water from Intensive Aquaculture of Fresh Water (Acronym: ABAWARE)

Stage 1 – Research on the use of microbiota as a basic method for selecting a new purification technology and the construction of a laboratory plant for wastewater treatment in aquaculture systems with recirculation based on selected microbial consortia
Activity 1.1 – Selection of Lemna sp. for the treatment of waste water resulting from SAR (recirculating aquaculture systems)

The steady increase of the global aquaculture sector from 49.9 million tonnes in 2007 to 66.6 million tonnes in 2012 (FAO, 2014) places it among the most dynamic sectors as development. The intensification of aquaculture production will continue to depend on the use of fishmeal and plant protein sources, mainly from soybean (Olsen and Hasan, 2012), which releases both large amounts of nitrogen and undigested phosphates in aquatic environments (Satoh et al. ., 2003, Nwanna et al., 2008). Nitrogen and phosphorus are the main nutrients in effluents from intensive aquaculture (Haghbayan and Mehrgan, 2015). Excessive phosphate in aquaculture effluents leads to the rapid spread of cyanobacterial infiltration, resulting in eutrophication in aquatic environments and the subsequent change in biodiversity structure (Kumar et al., 2011, Nwanna and Olusola, 2014). Ammoniacal nitrogen wastes, a limiting parameter of water quality in intensive aquaculture, are very toxic to macro-fauna in open water bodies (Lazzari and Baldisserotto, 2008). Stephen and Farris (2004) reported that increased ammonia concentrations may lead to blood ammonia intoxication or fish autointoxication.

In this context, intensification of aquaculture activity could also lead to a large amount of waste, including suspended solids, which results in a decrease in the dissolved oxygen concentration and finally the loss of aquatic biota (Magni et al. ., 2008). Reducing the production of these dissolved wastes is considered a key element for the long-term sustainability of aquaculture (Hasan, 2001). Duckweed (Lemna minor) has been used in several studies to absorb nutrients from aquaculture wastewater (Ansal et al., 2010). They are cheap and available, with the potential to eliminate nutrients after harvesting and therefore reduce the nutrient loading of the aquatic environment (Gupta and Prakash, 2014). The use of wastewater treatment and management makes it feasible for developing countries to provide low-cost household wastewater treatment, especially in rural areas (Smith and Moelyowati, 2001).

Table 1. Nitrogen and phosphorus removal in mg / m2 / day (after Reddy and de Busk, 1983)

Species Nitrogen mg/m2/day Phosphorus mg/m2/day
Eichhornia 1278 243
Pistia stratiotes 985 218
Hydrocotyle umbellata 365 86
Lemna minor 292 87
Spirodela polyrhiza 151 34
Azolla 108 33
Salvinia rotundifolia 406 105
Egeria densa 125 48

The Lemnaceae family includes Lemnaceae, a family of monocotyledonous aquatic plants with five genera and a total of 37 species: Spirodela, Lemna, Landoltia, Wolffia and Wolfiella, being considered as one of the highest growth angiosperms. All members of the Lemnaceae family are small, floating, freshwater plants, whose geographical areas extend across the globe. Lintida lives in freshwater ponds and basins, preferring waste water without current or slow current (Sree et al., 2016).


Rhinophyte species take up NH4 + and NO3 – both through their roots and the lower surface of their body (Lemna minor: Cedergreen and Madsen 2002), and may prefer NH4 + to NO3- (Landoltia punctata: Fang et al 2007). Higher concentrations of NH4 + in the environment are toxic to plants, animals and even humans (Britto and Kronzucker 2002). However, L. minor takes NH4 + slightly and increases well with ion concentrations of up to 84 mg / l (Zhang et al., 2014;

At higher concentrations of NH4 + leads to reduction of growth rate and loss of photosynthetic pigment. Their ability to pick up and tolerate relatively high levels of NH4 + makes it particularly suitable for the treatment of waste water from domestic and agricultural sources, which often contain considerable amounts of ion. More than 90% of NH4 +, 70% of NO3-, and 33% to 85% of PO4- present in Ghana’s wastewater (Awuah et al. 2004), domestically treated wastewater in Egypt (El-Shafei and et al., 2007) and domestic waste water from Israel (Ben-shalom et al., 2014) were eliminated by Spirodela polyrhiza, Lemna gibba / L. minor and L. gibba respectively.

These studies have shown that lignite waste water treatment has also maintained a neutral pH, reduced chemical and biological oxygen demand and eliminated suspended solids, mosquito larvae and coliform bacteria. Figure 1 shows the scheme of biological waste water processes using the lining.


 Fig. 1 Scheme of biological processes of wastewater using lignite, after M.D. Smith and Moelyowati, I. (2001)

The purpose of this research activity was to select Lemna sp. of the spontaneous flora and the determination of the species required for the treatment of waste water resulting from SAR. Figure 2 shows the selected Lemna species for the current study.

Figure 2 Selected Lemon Species

            Thus, for the experiments, 5 species of lobster were selected from local fauna: Lemna minor, L. trisulca, L. gibba, Wolffia arrhiza, Spirodela polyrhiza. Figure 3 shows the areas where the plant material has been tried and succeeded.

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Figure 3. Areas where the sampling of vegetal material has been tried and succeeded

            For the sampling we have established the aquatic habitat areas in which the plant species of interest could be found and we moved to several marshy areas around Bucharest (Văcăreşti Natural Park, Comana Natural Park, Snagov Forest Protected Area), but also in other parts of the country, such as the lower part of the Danube Delta Biosphere Reserve in the area bounded by the Lunar Lakes, Histria, Tuzla and Vadu Forest (Constanta County), along the Suha River and its affluent – the Branistea brook (Suceava County ), but we tested in the laboratory and the lingo already at Aquaterra Frasin, Suceava.


Figure 4 shows snapshots during sampling.

Species of Lemna were grown in tall pools and illuminated with fluorescent tubes. The intensity of illumination and the distance between the light source and the water level were tested to see the favorable conditions for growing the species of interest.

Several conclusions of the experiments on the growth of Lemna species under laboratory conditions:

  • In experimental vessels we noticed that the most favorable light for Lemna crops at a water depth of 25-35 cm) is 36 watts. Virtually one neon tube of that power is enough.
    •    If we use a 2-tube lamp, there is already too much light in the filtration system, and green algae and even worse algae and cyanobacteria that compete with the Lemna population are starting to develop and hinder the exuberant development of Lemna crops.
  •  The ideal neon tube distance to water is about 30 cm. Placing them at a smaller distance leads to the development of cyanobacteria.
  •  If we have a permanent recirculation system with a small recirculation pump that does not lead to the Lemna sp plant overturning, the plant growth rate increases.
    •   The length of Lemna minor roots is about 3-5 cm in poor crops in organic matter, especially if the aquaculture environment does not have fish. On the recirculating systems of our measured average experiments of Lemna minor roots, it reaches 10-12 cm in length, which is even thicker and more vigorous.
  •  Cultivation should be done as much as necessary, but with 90% of the surface area of ​​the aquaculture area occupied with Lemna, in order not to create a dangerous break for algal species, especially cyanobacteria (Mycrocystis aeruginosa, Nostoc sp.).
  •  At the same intensity of light, Lemna gibba plants develop very well, having the capacity to divide at least 2 times larger than the Lemna minor, which we have not found in the specialized literature. So he can be a good actor for filtering and recirculating water from RAS from superintensive aquaculture farms.
  •  In some experimental vessels we cultivated Lemna trisulca, the only species that lives both on the surface and in the water. We have experienced the cultivation of this species in 3 experimental variants:
  1. on Ceratophyllum submersus plants – as we have found them associated in the natural environment of the Gurban Valley – Comana Natural Park.
  2. on a rabbit mesh network parallel to the slats frames to be well stretched. These bars are placed parallel to each other. Woven plants as a network develop very well if the water comes with enough organic substance from the aquaculture basins of the fish. The lifecycle must be permanently reophilized by a recirculation pump (brook-type micromedium or slow flow but constant flow filter).
  3. High-grain filtering polyurethane foam or special polyurethane foam strands used for retaining as much organic deuterium as specially used in aquaculture recirculating systems.
  •  In all systems used in experiments we used a higher intensity of light (for experimental vessels we used a light intensity of 4 tubes of 18watts at a water depth of about 30 cm of water. higher current than the other species of surface Lemna and the ecological conditions thus created provide the plant with harmonious conditions of development according to their ecological requirements. The lateral illumination can be provided by a proposed new type of long-life aquarium with vertical illumination lamps and two lateral lighting systems in such a way that the Lemna trisulca filter population grows illuminated from all sides. After filtration the water has to be sterilized with a UV lamp – before it returns to the aquaculture environment.

ISI articles and conferences on the project

ISI articles:

  •  Ioana Corina Moga, Nicolae Crăciun, Ioan Ardelean, Gabriel Petrescu, Radu Popa, THE POTENTIAL OF BIOFILMS FROM MOVING BED BIOREACTORS TO INCREASE THE EFFICIENCY OF TEXTILE INDUSTRY WASTEWATER TREATMENT, The Publishing House of Industria Textilei Magazine, undergoing printing, acceptance letter in the figure below:



Conferences / Dissemination activities:

  •  Participation in Euroinvent 2017 – European Exibition of Creativity and Innovation, 25-27 May 2017 and gold medal at this occasion

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Certificate of participation and prize



  •  International Conference Tex Teh VIII, 19 -20 Octombrie 2017, București, presentation and title article: ”THE POTENTIAL OF BIOFILMS FROM MOVING BED BIOREACTORS TO INCREASE THE EFFICIENCY OF TEXTILE INDUSTRY WASTEWATER TREATMENT”


Snapshot from project submission

  •  International Symposium ISB – INMA TECH, Agricultural and Mechanical Engineering, 26 – 28 Octombrie 201


Participation Diploma
•    13th International Symposium “Priorities of Chemistry for Sustainable Development” – PRIOCHEM, 25-27 October, Bucharest, venue: UPB Central Library with poster and paper: AMS-CARRIER-CHLORELA BIOFILM IS USEFUL IN AQUACULTURE WASTEWATER MANAGEMENT

Bibliografie selectivă

Ansal M D, Dhawan A and Kaur V I 2010: Duckweed based bio-remediation of village ponds: An ecologically and economically viable integrated approach for rural development through aquaculture. Livestock Research for Rural Development. Volume 22, Article #129. Retrieved December 4, 2017, from http://www.lrrd.org/lrrd22/7/ansa22129.htm
Asada, K. (2006), Production and scavenging of reactive oxygen species in chloroplasts and their functions. In: Plant Physiology, vol.141, nr. .2, 391-396.
Bidlack, J.E.; Stern, K.R., Jansky, S. (2003). Introductory Plant Biology. New York: McGraw-Hill
Cedergreen N,  Madsen TV. Light regulation of root and leaf NO 3− uptake and reduction in the floating macrophyte Lemna minor, New Phytologist , 2003, vol. 161 (pg. 449-457)
Fiske CH, Subbarow Y. 1927. The nature of the “inorganic phosphate” in voluntary muscle. Science. 65(1686):401-403
Haghbayan, S., and M. S. Mehrgan. 2015. The effect of replacing fish meal in the diet with enzyme-treated soybean meal (HP310) on growth and body composition of rainbow trout fry. Molecules 20: 21058–21066
Hasan, M.R. Nutrition and feeding for sustainable aquaculture development in the third millennium. in: R.P. Subasinghe, P. Bueno, M.J. Phillips, C. Hough, S.E. McGladdery, J.R. Arthur (Eds.) Technical Proceedings of the Conference on Aquaculture in the Third Millennium, Bangkok, Thailand, February 20–25. NACA/FAO, Bangkok/Rome; 2000:193–219.
Konda Ramesh Reddy, W. F. De Busk, Nutrient Removal Potential of Selected Aquatic Macrophytes, October 1985Journal of Environmental Quality 14(4)
Kumar, V., A. K. Sinha, H. P. S. Makkar, G. De Boeck, and K. Becker. 2011. Phytate and phytase in fish nutrition. Journal of Animal Physiology and Animal Nutrition 96: 335–364. doi:10.1111/j.1439-0396.2011.01169.x.
Lazzari R, Baldisserotto B (2008) Nitrogen and phosphorus waste in fish farming. Bol Inst Pesca 34(4):591–600
Magni, P., Rajagapal, S., Vandervelde, G., Perel, G., Kasserberg, J., Vizzini, S., Mazerla, A. and Giordanb, G. (2008). Sediment features, macroziobathic assemblages and trophic relationship following a dystrophic event with anoxia and sulphide development in the Santa Giuta Lagoon. Marine Pollution Bulletin 57:125 – 136.
Nwanna LC (2003). Nutritional Value and Digestibility of Fermented Shrimp Head Waste Meal by African Catfish Clarias gariepinus. Pak. J. Nutr. 2: 339-345.
Olsen, R.L. and Hasan, M.R. 2012. A limited supply of fishmeal: Impact on future increases in global aquaculture production. Trends in Food Science and Technology, June 2012.
Olusola and Nwanna, 2014, Growth Performance of Nile Tilapia (Oreochromis niloticus) Fed Processed Soybean Meal Based Diets Supplemented With Phytase, International Journal of Aquaculture, Vol.4, No.08: 48-54 (doi: 10.5376/ija.2014.04.0008)
SATOH, S.; HERNÁNDEZ, A.; TOKORO, T.; MORISHITA, Y.; KIRON, V.; WATANABE, T. 2003 Comparison of phosphorus retention efficiency between rainbow trout (Oncorhynchus mykiss) fed a commercial diet and a low fish meal based diet. Aquaculture, Amsterdam, 224:271-282.
Sree, K. S., Bog, M., & Appenroth, K.-J. (2016). Taxonomy of duckweeds (Lemnaceae), potential new crop plants. Emirate  Journal  of  Food  and  Agriculture,  28, 291–302.
Stephen, W.W. and Farris, J.L. (2004). In stream community assessment of aquaculture effluents. Aquaculture 231:148-162.
Smith, M.D., Moelyowati, I., 2001. Duckweed based wastewater treatment (DWWT): design guidelines for hot climates. Water Sci. Technol. 43 (11), 291–299.

Management Team

Philip Hoffman

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David Spencer

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Richard Samuels

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Anissa Doel

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