Cultivation of microalgae in industrial wastewaters

Microalgae production for the purpose of clearing wastewater has been researched for at least half a century. Such systems have a dual benefit: first, they prevent nutrients from entering water bodies and causing eutrophication; second, they transform sunlight and carbon dioxide into a biomass that has many potential uses.

Unfortunately, the current high costs of cultivation have limited the development and exploitation of such systems, resulting in only a few full-scale algae wastewater treatment installations and a small industry based mostly around food and pigments. This thesis contributes to a growing body of knowledge with the aim to make algae cultivation viable for the production of sustainable products.

Specific contributions include: improvement in the methods of screening the growth potential of different microalgae species; identification of an industrial wastewater that allows good algae growth; knowledge about the mixotrophic utilization of chemical energy present in organic waste; demonstration of a method to optimize efficiency of culture growth and nutrient removal; and biochemical characterization of the produced biomass.

When designing algae cultivation, one challenge is that there are many potential combinations which must empirically screened. Tens of thousands of microalgae species have been identified so far and there are numerous waste-streams that potentially could be of interest. A screening system was developed using the microplate as cultivation vessel and measurement cuvette.

Fluorescence was demonstrated to be an order of magnitude more sensitive than optical density for detecting biomass growth, which increased the length of time in which exponential growth was observable from hours to days. This enabled growth rate-light intensity (µ-I) curves to be measured in microplates which were found to be equivalent to those obtained in typical lab-scale photobioreactors.

As µ-I curves are the key biological input to an already existing model, it was validated that low density microplate cultivations can be used to make predictions about industrially relevant autotrophic cultivation. When algae are grown within a wastewater treatment plant, the use of the chemical energy stored in the organic carbon dissolved in the wastewater could also be a useful option.

Conventional aerobic sewage treatment expends much energy in breaking down the biomass to CO2. However, various anaerobic treatment methods would result in effluent containing dissolved organic molecules suitable for algae species that have the ability to grow as mixo- or heterotrophs. Chlorella sorokiniana was cultivated in a lab scale photobioreactor under daily light dark cycles and various timing strategies were tested for adding acetate at concentrations that can be obtained in waste streams of 1 – 2 g L-1.

The results showed that the fastest growth occurred when adding the acetate at night (cyclic autotrophy/heterotrophy). However adding the acetate during the day (mixotrophty) also improved growth compared to autotrophic controls. Industrial wastewater was used as cultivation medium of Chlorella sorokiniana.

The culture was able to grow at high rates upto a density of 4 g L-1. The deceleration-stat technique was used to create a series of pseudo-steady states to give information about the expected results of continuous cultivation of microalgae in the selected wastewater. At light intensities of 2100 and 200 µmol photon m-2 s-1 the algae grew at a rate of over 5 and 1.67 g L-1day-1, respectively.

The corresponding removal rates of nitrogen were 238 and 93 mg L-1day-1 and 40 and 19 mg L-1day-1 for phosphorous. Ammonium removal varied from below 40% to 99%, while phosphate removal was always nearly total. When the biomass was characterized, it was found that fertilizer value N and P content increased with growth rate.

For animal feed, the amino acid content was about 40% of biomass. The content of the nutritionally important α-Linoleic fatty acid increased when light intensity and dilution rate were higher. Valuable pigments lutein, carotene and other carotenoids were higher in low-light conditions. The results from this thesis demonstrate that industrial wastewater can be a suitable replacement for algae cultivation medium.

The screening method developed will reduce the cost of identifying the best conditions to test at lab scale. The D-stat method offers a way to identify the best conditions for biomass production and nutrient removal. Various options for heterotrophic and mixotrophic utilization of waste organic carbon in effluents are identified.

Further advances in microalgae cultivation and processing will be needed for the production of sustainable products from wastewater in the future.