Slide 3.

Introduction Problem:
What is the problem?

Conventional swimming pools normally use chlorine as a residual disinfectant. While the bacteria-killing properties of chlorine are very useful, chlorine also has some side effects that can be annoying to humans, and possibly even hazardous. Those are caused by the formation of Disinfection By Products (DBP), which are mostly composed of chloramines and trihalomethanes. Several studies have shown a relation between DBPs exposure and health effects such as skin and eyes irritation and also increased risks of cancers.

Slide 4. Introduction-Project
So, the solution is a residual disinfectant free swimming pool. Some companies have developed alternatives to chlorine, including other chemicals and ion generators. Some of these are good alternatives, but they don't achieve the cleanliness, oxidation levels or low price that chlorine provides. But theres always a possible problem while looking for this utopic pool, and its the biofilm formation.

Slide 5. Introduction What is a biofilm?

Biofilm is any group of microorganims in which cells stick to each other. Normally there are 5 stages in the biofilm formation. 1. Initial attachment: this stage is carried on when the first microorganisms attach themselves to the wall, this is a weak attachment. 2. Irreversible attachment: If the first colonists are not immediately removed then they can be strongly attached to the walls via their Extra Polymeric Substances or EPS. 3. Maturation I: The production of these polysaccharides that anchor the bacteria to the surface allow colonies to grow. 4. Maturation II: replication and recruitment. 5. Dispersion: dispersal of cells from the biofilm colony is an essential stage of the biofilm life cycle because this stage enables the biofilm to spread and colonize new surfaces.

Slide 6. Introduction UV.

This biofilm needs to be eliminated from the swimming pools environment because biofilm have been found to be involved in a wide variety of microbial infections, by one estimate 80% of all infections such as: urinary tract infections, catheter infections, middle-ear infections, formation of dental plaque, gingivitis and so on. One way to avoid the problem of the biofilm growth is to eliminate it with UV light. UV light has proven to be an effective method. There is one wavelength in the UV range that is specially used for sterilize, its situated at 254 nm. How it works? UV can penetrate through the cell membrane of a microorganism and block deoxyribonucleic acid (DNA) replications, which eventually results in lethal damage.

Slide 7. Methods and Materials Set-up.

The biofilm incubator has 2 zones, in each zone it has 2 lines and finally each line has 6 PVC jars, which are representing little pools. Then, totally it has 24 jars of 140 ml capacity each one. Each jar is connected to the previous one by a tube and to make the flow easier and constant each jar is elevated by a wood piece, each piece is 0,8 cm lower than the previous one, making a ramp. The flow it is set to 480 ml/h of tap water and 20 ml/h of BFA and the temperature is set to 33C in the environment to reach 32C in the water. BFA is an acronym of Body

Fluid Analogs (BFAs) were used to simulate the import of pollutants by the bathers, mostly urine and sweat.

Slide 8. Methods and Materials Experiments.

Four experiments were performed and 2 parameters related with UV were varied. As we can see, those parameters are the days/week of application and the UV dose. In the first experiment, no UV was applied. And in the forward experiments the UV was applied beginning with a small dose and intensifying this those from experiment to experiment.

Slide 9. Methods and Materials Analysis.

The cATP is measuring the cellular ATP concentration, which can be related to the total bacteria present in each jar The chemicals give some information about the consumption and transformation of medium by the microorganisms.

Slide 10 and 11. Results and discussion Experiment 1.

This graph corresponds to the average quantity of biofilm in each day of sampling. Theoretically a biofilm population should grow every day so the quantities should be higher as time passes, and as we can see there was a problem with our biofilm development. It was a FLOW PROBLEM. If the flow is not constant then, the fresh water with the nutrients is not reaching the jars and the biofilm cannot grow anymore. Thats what happened in 2nd and 4th days of sampling. How we know? Easy! Urea and phosphate are the main nutrients for the biofilm and if theres a defficience in Urea or phosphate the biofilm development cannot proceed. So, biofilm begins to die.

Slide 12. Results and discussion UV Experiment.

This graph corresponds to For experiment 2, UV was applied 24h before days 7 and 14. And on experiment 3, UV was applied 24h before every day of sampling. For the first days of sampling, day 4 and day 7, the quantities are almost the same for both experiments because theres not enough biofilm growth to see the difference. For day 11 of sampling we can see that in Experiment 3 theres an appreciable difference between the lane where the UV was applied and the lane where UV wasnt applied. Theres a decrease in the quantity of biofilm. This decrease is not appreciable in Experiment 2 because no UV was applied the day before. Finally, for the last day of sampling, in Experiment 2 theres a huge difference, around 50% of biofilm decrease, between the lane with UV and the lane without UV. And for Experiment 3, theres a smaller decrease but this is because in Experiment 3 UV was applied every day so there was less total quantity of biofilm to kill.