1. Tests performed on By Corporate Marketing Department - Tecniplast, Buguggiate -

2. Pressure – Airflow relationship Aim: Find the relationship between set pressure and intra-cage ACH (ISON36). Method: Record the exhaust airflow and compute cage ACH, having set the system to -75, -100 and -125 Pa. Repeat the measurement after removing 18 and 34 cages. Results: Findings: With all the cages in place P set point (Pa)Cage ACH -7553 -10064 -12579 With only 2 cages inside rack, the internal pressure is constant and the pressure is the same of the set point The increase of the exhaust airflow is caused by the air sucked from the exhaust nozzles of the cages removed In the positions of the cages removed the supply nozzles get closed and in the remaining cages pressure and ACH are constant

3. Pressure distribution Aim: Evaluate the homogeneity of the intra-cage pressure across an ISO72N rack. The goal is to stay within ± 10 %. Method: Settings: -100 Pa. Record the intra-cage pressure in key positions through a pressure gauge. Results: Findings: Recorded pressure are within ± 10 % range frontback

4. Backup battery duration Aim: Evaluate the battery lifetime in case of power failure. Method: Settings: +100 Pa (test performed with ISOcage P system) Switch off the electricity supply Record the electrical tension of the batteries during time Minimal tension required for batteries: 21,6 V (useful to recharge batteries) Results: Findings: Battery lifetime > 48 h tested on a 72 cage places rack, with brand new battery 22.9 V @ 48 h

5. Noise Level Aim: Evaluate the noise emitted by an ISO36N. The goal is to stay within below 50 dB. Method: Settings: - 25 Pa; -100 Pa; -125 Pa. Record the noise 1 meter away from the AHU. Results: Worst case: Findings: The noise level emitted by the AHU was below 50 dB even in the worst case (working at -125 Pa)

6. Static conditions Aim: Evaluate the intra-cage environmental conditions when the cage is not ventilated Method: ISOcage with 5 CD1 mice (>50g) 120g of bedding Detection for 15 minutes of CO2 NH3 O2 CH4 Detection of T and RH for 30 minutes Results: Findings: CO2 overtake the max. accepted level in 15 minutes.

7. Intra-cage pressure with just one blower working Aim: Validate if the system is able to reach and mantain the set point even if one of the two blower is not working. Method: Settings: -75 Pa, -100 Pa, -125Pa. Intra cage pressures are measured. Results: Findings: Set point was mantained

8. Validation of decontamination rack Aim: Verify the that autoclave cycles is effective, acting through the opening of the HEPA protected rear valve of the ISOCAGE by means of the ISO decontamination rack. Method: - Three ISOcages, complete with bedding and food. - Four spore strips of 1.9x 106 of Geobacillus stearothermophilus - Autoclave cycle, 121°C – 20 min Results: All cultures of the spore strips were negative after 7 days of incubation at 55° to 60°C. Findings: The possibility to sterilize safely the “ISOCAGE hermetic enclosure” by means of a simple device like the decontamination rack was demonstrated.

9. Acceptance tests report These tests are performed in 100% of manufactured AHUs: DOP test – filter integrity test Aeraulic test Electical tests Sensor calibration