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The reliability of a robust field incubator lies in its ability to reach and maintain a set temperature under various conditions. To monitor the performance of the various incubator set-ups, the following measurements were made: time needed to reach the set temperature, effect of opening the door for one minute, power consumption over 24 hours of operation, inner temperature stability over 24 hours of operation, and observation of E. coli growth. The temperature inside the incubator was measured every minute with 4 temperature logging devices placed in different positions in the structure (support rack, wall, top, inside a growth plate). The set temperature was considered to be attained when all measurements were within plus or minus 2 °C, which is the acceptable range for the incubation of E. coli.13
The electronic core was tested with three types of shells, using materials that are typically found in many countries: a polystyrene foam box (78 liters), a hard plastic cooler box (30 liters), and a cardboard box covered with a survival blanket (46 liters) (Figure 10). To cover a range of ambient conditions that can be experienced in the field, these incubator set-ups were tested at three ambient temperatures: ambient (about 27 °C), cold (about 3.5 °C and 7.5 °C) and hot (about 39 °C). Performance measures were tested setting the inner temperature at 37 °C and 44.5 °C.
The time to reach the set temperature in the incubators was influenced by the ambient temperature and the material of the incubator shell. At an ambient temperature of about 27 °C, the three incubators set-ups reached the set temperatures (37 °C and 44.5 °C) in a similar time (Figure 11a and Figure 12a) and comparable with the performance of a standard incubator (Table 3). In cold environments (3.5 °C and 7.5 °C), the incubators with thicker shells, i.e., the polystyrene foam and cooler box, reached the target set temperatures (37 °C and 44.5 °C) in a similar time; about four times longer than under an ambient temperature of 27 °C. With its lower insulation, the cardboard box with survival blanket never fully reached the set temperatures under cold ambient temperature conditions (Figure 11b and Figure 12b). In a warm environment (39 °C), the three incubator set-ups reached the target temperature of 44.5 °C in under 10 minutes (Figure 12c). However, when the set temperature was of 37 °C, i.e., lower than the ambient temperature, none of the incubators could lower the temperature, resulting in overheating for all three incubator set ups (Figure 11c).
The ambient temperature and the type of incubator shell influenced the impact of opening the door of the incubator for one minute. The heat loss was greater in the cold environment, and the time to recover the inner set temperatures was longer, with the exception of the cardboard box incubator where the set temperatures were never reached (Figure 13b and Figure 14b). In the warmer environments, the heat loss was limited and the set temperatures were recovered in under 10 minutes (Figure 13a,c and Figure 14a,c). In an ambient temperature of 39 °C and set temperature of 37 °C, opening the door did not cause nor reduce overheating of the incubators (Figure 13c).
The power consumption increased with cold environments and with an increase in the set temperature. Better insulating incubator shells (polystyrene foam and cooler box) showed a reduced power consumption as compared to the cardboard box incubator. In similar environments (ambient temperature of about 27 °C), the three incubator set-ups consumed 0.22 to 0.52 kWh/24h less energy than the standard incubators tested (Table 3).
The temperature in the incubator remained stable over 24 hours with all type of incubator shells and ambient temperature tested (Figure 13 and Figure 14). Slight variations of the measured temperature compared to the set temperature were observed according to the position of the temperature logging device in the incubator. With the exception of the tests with the ambient temperature (39 °C) warmer than the set temperature (37 °C) (Figure 13c), the variations in temperature were all within the 2 °C acceptable range for E. coli incubation.
All tests were performed in the presence of E. coli and total coliform measurement materials (membrane filter placed on growth plate). Replicates of a sample were placed in each incubator set-up and in a standard incubator for comparison. In all set-ups and conditions, the growth of E. coli and total coliform was successful and comparable to the growth observed in the standard incubator. A summary of the incubator configurations and ambient temperature conditions tested with results are shown in Table 3.
| | | Test 1:
Time to set temperature | Test 2:
Side door opening one minute | Test 3:
Power consumption over 24-hour period | Test 4:
Temperature variation over 24-hour period | Test 5:
E. coli growth observed |
| Ambient temperature | Set temperature | (min) | Maximal loss of temperature (°C); time to recover set temperature (min) | (kWh/24 h) | Absolute maximal temperature (°C); absolute minimal temperature (°C) * | (Yes / No) |
| Polystyrene foam box | 3.5 °C | 37 °C | 45 | 10 °C; 17 min | 0.78 | 37; 35.5 | Yes |
| 7.5 °C | 44.5 °C | 74 | 16.5 °C; 31 min | 0.89 | 44.5; 42.5 | ND† |
| 27 °C | 37 °C | 12 | 2.5 °C; 3 min | 0.28 | 37.5; 36.5 | Yes |
| 44.5 °C | 20 | 4.5 °C; 7 min | 0.43 | 44.5; 43.5 | ND† |
| 39 °C | 37 °C | 0 (overheat) | 2 °C; 0 min (overheat) | 0.11 | 42.5; 42 | Yes |
| 44.5 °C | 7 | 3.5 °C; 5 min | 0.17 | 45; 43.5 | ND† |
| Hard cooler box | 3.5 °C | 37 °C | 54 | 8 °C; 10 min | 0.86 | 37.5; 36 | Yes |
| 7.5 °C | 44.5 °C | 96 | 12 °C; 30 min | 1.05 | 45; 43 | ND† |
| 27 °C | 37 °C | 13 | 1.5 °C; 0 min | 0.27 | 37.5; 36.5 | Yes |
| 44.5 °C | 25 | 2 °C; 4 min | 0.50 | 45; 43.5 | ND† |
| 39 °C | 37 °C | 0 (overheat) | 1 °C; 0 min (overheat) | 0.11 | 43; 42.5 | Yes |
| 44.5 °C | 9 | 4 °C; 3 min | 0.19 | 45.5; 44.5 | ND† |
| Cardboard box with survival blanket | 3.5 °C | 37 °C | Never reached (stable temperature after 109 min) | 6.5 °C; stable temperature after 30 min | 1.24 | 33.5; 30.5 | Yes |
| 7.5 °C | 44.5 °C | Never reached (stable temperature after 120 min) | 8 °C; stable temperature after 20 min | 1.28 | 36.5; 32 | ND† |
| 27 °C | 37 °C | 15 | 2.5 °C; 6 min | 0.42 | 36.5; 35.5 | Yes |
| 44.5 °C | 24 | 3 °C; 8 min | 0.70 | 44.5; 42.5 | ND† |
| 39 °C | 37 °C | 0 (overheat) | 1.5 °C; 0 min (overheat) | 0.11 | 41.5; 40 | Yes |
| 44.5 °C | 9 | 2 °C; 0 min | 0.20 | 45; 43.5 | ND† |
| Standard incubator | 27 °C | 37 °C | 18 | 1 °C; 0 min (overheat) | 0.64 | 38.5; 36 | ND† |
| 44.5 °C | 23 (overheat) | 2.5 °C; 0 min | 0.95 | 47.5; 43.5 | ND† |
Table 3: Results summary for the incubator configurations and ambient temperature conditions tested. *Test 4: Absolute maximal and minimal temperatures recorded during the stable periods, i.e., from 10 minutes after the end of a disruptive event (time to reach set temperature, opening the door). †ND: No data, test not run.

Figure 10: Incubator shells tested. Open (upper row) and closed (bottom row). Polystyrene foam box (left), thickness of 3.5 cm, outer dimensions 39 x 56 x 36 cm; hard plastic cooler box (middle), thickness of 2.5 cm, outer dimensions 32 x 41 x 47 cm; cardboard box (right) covered with a standard survival blanket of a thickness of 12 µm folded twice, outer dimensions 30 x 42 x 37 cm. Please click here to view a larger version of this figure.

Figure 11: Time to reach set temperature (37 °C) of the incubator set-ups under different ambient temperature conditions. Performances of incubators with a shell made of a polystyrene foam box, a hard cooler box, and a cardboard box covered with a survival blanket. At room ambient temperature (a), cold ambient temperature (b), and warm ambient temperature (c). Temperatures recorded on the support rack of the incubators. Please click here to view a larger version of this figure.

Figure 12: Time to reach set temperature (44.5 °C) of the incubator set-ups under different ambient temperature conditions. Performances of incubators with a shell made of a polystyrene foam box, a hard cooler box, and a cardboard box covered with a survival blanket. At room ambient temperature (a), cold ambient temperature (b), and warm ambient temperature (c). Temperatures recorded on the support rack of the incubators. Please click here to view a larger version of this figure.

Figure 13: Temperature variations over 24-hour period and effect of door opening under different ambient temperature conditions. Set temperature of 37 °C. Performances of incubators with a shell made of a polystyrene foam box, a hard cooler box, and a cardboard box covered with a survival blanket. At room ambient temperature (a), cold ambient temperature (b), and warm ambient temperature (c). Circled areas show the temperature variations due to the door opening for one minute. Temperatures recorded on the support rack of the incubators. Please click here to view a larger version of this figure.

Figure 14: Temperature variations over 24-hour period and effect of door opening under different ambient temperature conditions. Set temperature of 44.5 °C. Performances of incubators with a shell made of a polystyrene foam box, a hard cooler box, and a cardboard box covered with a survival blanket. At room ambient temperature (a), cold ambient temperature (b), and warm ambient temperature (c). Circled areas show the temperature variations due to the door opening for one minute. Temperatures recorded on the support rack of the incubators. Please click here to view a larger version of this figure.

Figure S1: Electrical diagram of incubator electrical core wiring. Alternatives for mains operation and battery operation are indicated. Please click here to view a larger version of this figure.
| Parameter | Selected value |
| 1 | Type of control output | Control Q1 / Alarm Q2 |
| 2 | Type of connected sensor | Pt100 (-200 to 140°C) |
| 3 | Lower limit selectable for setpoint value | 0 |
| 4 | Upper limit selectable for setpoint value | 50 |
| 5 | Type of control | Heating |
| 6 | ON/OFF hysteresis or dead band for P.I.D. control | 0 |
| 7 | Proportional bandwidth of the process expressed as units (°C if temperature) | 1 |
| 8 | Integral time. Intertia of the process expressed as seconds | 80.0 |
| 9 | Derivative time for P.I.D. | 20.0 |
| 10 | Cycle time for time-proportioning output | 10 |
| 11 | Allow/deny modification of setpoint values by frontal keyboard | Allow modification of all setpoints |
| 12 | Software filter. Number of readings to calculate the comparison value PV-SPV | 10 |
| 13 | Type of degree | °C |
| 14 | Type of cooling liquid | Air |
Table S1: PID temperature controller settings. Display of set values; other parameters not necessary to run the incubator were left to default values.