TITLE: WATER PRODUCTION FOR AWARE (Total Coliforms, Fecal Enterococci and Escherichia coli:)
AUTHORS: Celia Manaia

[DESCRIPTION]
The protocol summarises the procedures used for analytical control. The protocol describes the
Standard Operating Procedure (SOP) for the optimization of advanced tertiary treatment of water, based on a comprehensive quality and risk assessment.

[GUIDELINES]
RECOMMENDED/ACCEPTED VALUE:

Total coliforms: Amount0 MPN/ mL  
E.coli: Amount0 MPN/ mL   
Fecal enterococci: Amount0 MPN mL   

[STEPS]
SECTION: Methods: The section below summarises the procedures used for analytical control – detailed protocols are annexed to this protocol.
2. Total Coliforms, Fecal Enterococci and Escherichia coli: 
SECTION: Methods: The section below summarises the procedures used for analytical control – detailed protocols are annexed to this protocol.
2.1. Analysis: Enumeration of culturable mesophilic bacteria at37 °C 

SECTION: Parameters framed by Legal and Regulatory Requirements:
3. Using the EU Drinking Water Directive:  

Mesophilic Bacteria in PCA (Plate Count Agar) – 0 CFU/100 mL

Total coliforms and Escherichia coli –Number /100 mL (0 MPN/100 mL) 

Fecal enterococci –Number/100 mL (0 MPN/100 mL)  

Viral concentration - There are no legal requirements for viruses. They are not included in any regulation now. 

Parasite  - EU legislation (2020/741)  

Metals - DIRECTIVE 2008/105/EC OF THE EUROPEAN PARLIAMENT AND OF THE COUNCIL of 16 December 2008 on environmental quality standards in the field of water policy 

Organic contaminants - DIRECTIVE 2008/105/EC OF THE EUROPEAN PARLIAMENT AND THE 
COUNCIL of 16 December 2008 on environmental quality standards in the field of water policy.  

SECTION: WATER PRODUCTION FOR AQUAPONICS
1. The water production for AWARE main activities includes three stages – disinfection by ultraviolet C radiation (UVC), storage for720 min-1440 min(according to water load and season) and ozonation. The water quality is monitored at these three stages, for the parameters indicated in Figure 1 below.

 

SECTION: WATER PRODUCTION FOR AQUAPONICS
1.1. Sampling, Processing, and Analyses 

Water samples are collected (see Figure 2) and processed within a360 mininterval, before being shipped for the partner responsible for the analyses (Table 1). In case no processing is needed, samples are frozen and stored at-80 °Cwithin180 min.
For each sampling event, the date, day of the week and hour; the temperature and rain. Sampling points, indicated in Figure 2 were designated from A to I:
- Influent of primary treatment (A) 
- Influent of biological treatment (activated sludge) (B) 
- Treated secondary effluent (C) 
- Sand filter effluent (D) 
- UVC effluent (E) 
- Storage for reuse tank effluent (F) 
- Ozonation effluent (1 dose, e.g., 5 mgO3) - MITO3X technology - (G) 
- Effluent of the vacuum UV oxidation (VUV) (H) 
- Effluent of reactive storage / Influent of the recirculation aquaculture system (RAS) (I) 

 

SECTION: Methods: The section below summarises the procedures used for analytical control – detailed protocols are annexed to this protocol.
2.2. Method:   
 
Jspreadsheet CE
Reference:Go togo to step #2.3   
− UNI EN ISO 9308-2:2014 Water quality - Count of Escherichia coli and coliform bacteria - Part 2: Most probable number method. 
− EN ISO 11133:2014 - Microbiology of food, animal feed and water. Preparation, production, storage and performance testing of culture media. 
 
Jspreadsheet CE
Reference:Go togo to step #2.3   
− UNI EN ISO 7899-1:2001 - Water quality - Research and enumeration of intestinal enterococci - Miniaturized method (Most Probable Number) for surface and wastewater. 
− EN ISO 11133:2014 - Microbiology of food, animal feed and water. Preparation, production, storage and performance testing of culture media. 
SECTION: Methods: The section below summarises the procedures used for analytical control – detailed protocols are annexed to this protocol.
2.3. Observations: Samples were processed and analysed withinDuration 720 min  after collection.