126

CHAPTER 7

CONCLUSION

7.1

SUMMARY

The aim of the present study was to explore the potential of PEF as
an emerging non-thermal method for liquid food preservation. The efficiency
of the process is dependent on a number of parameters unique to the
technology. The effect of the process, product and microbial parameters were
established by Finite Element Analysis and validated with experimentation.
The conclusions arrived at each step are elaborated below by highlighting the
salient findings.

7.2

CONCLUSIONS

7.2.1

Electrical field modeling of bacterial cell

The isolated bacterial cell was modelled using a FEM package.

Effort was made to estimate the effect of orientation angle on TMP for
successful permeabilization of the cell membrane. The following conclusions
were drawn.

The critical electrical field for microbial inactivation with

TMP in range of 0.72 to 2.2 V is 1.5 kV/cm.

The effect of orientation angle on the Ec revealed that Ec is

maximum (3.1 kV/cm) along the major axis, which supports
earlier literature and validated by analytical method.

127

For every 30 increase in the orientation angle the TMP

decreases at a rate of 5.5 V.

The critical electric field increases with an increase in the

applied electric field, with maximum of 41% of the applied
field.

The exact membrane capacitance was found to 1.226 F / cm 2

against 1 F / cm 2 as stated by TsaiFuwu (2004.)

For a given field strength, the energy and capacitance of the

bacterium remains almost constant for all orientation angles.

7.2.2

Effect of critical parameters on PEF processing of liquid foods

The FEM model of the bacterial cell was used for estimating the
dependence of Ec on product factors, process factors and microbial factors etc.
by modeling the treatment chamber varying the liquid mediums.

Though contradictory results are cited in the literature

regarding the dependence of microbial inactivation on the
conductivity of the medium, milk with a conductivity of 0.5
S/m requires 3.15 kV/cm to induce TMP greater than the
threshold when compared to fresh water of conductivity 0.01
S/m which requires 2.6 kV/cm. The conductivity causes a
difference in the ionic strength in the medium and cytoplasm
that weakens the membrane making it more susceptible to an
electric pulse.

128

The dielectric constant of the food has a significant influence

on the applicability of PEF. The Ec decreases with an increase
in the dielectric constant.

The electrode geometry and spacing between the electrodes

also influence inactivation. Non-uniformity demands lesser
average field for inactivation when compared to uniform
fields.

Microbial factors namely shape and pre-existence of pores on

the membrane influence the Ec significantly.

Nearly 25%

more field strength is required under uniform field

configuration for spherical cells with pores in comparison to
ellipsoidal cells.

Fat acts as an insulator and thus the milk with higher fat
content requires higher field.

Effective permeabilization thus requires high medium

conductivity, low fat content and also needle shaped
electrodes with less spacing.

Thus the FEM model developed forms an essential tool in

improving the efficiency of the PEF process.

7.2.3

Effect of input pulses on PEF processing

The feasibility of different waveforms for electroporation was

studied by PSPICE simulation using the electrical equivalent model of the
bacterial cell and the findings are reported herewith.

129

An exponentially decaying impulse voltage waveform is an

optimum source for liquid food preservation.

The simulations reveal that impulse voltages can effectively

inactivate microbes and electric field intensity is a vital factor
for successful sterilization. The electrical field in the range of
15 kV/cm 80 kV/cm is ideal for liquid food preservation

The number of pulses has a detrimental effect on the lower

electric field when compared to the higher field. The
minimum threshold for the number of pulses to be applied for
PEF analysis is 10 below which significant reduction is not
seen, but this number varies with field intensity

Circuit inductance mainly affects the shape of the input

waveform by reducing the peak voltage and current applied to
the voltage treatment chamber

When the circuit inductance is high, the pulse has a slow

rising waveform with increased oscillations

With the high conductivity of the liquid foods, any small

changes in the circuit inductance has a substantial influence on
the measured pulse

Knowing the conductivity of liquid food to be treated with a

realistic estimation of a circuit inductance, it is possible to
predict the shape of the pulse under different test conditions.

The surveillance calculator forms a supportive tool in

optimizing the process parameters aiding for a complete
feedback control system for efficient PEF process.

130

7.2.4

PEF analysis on liquid food

The efficacy of pulsed electric fields in food preservation was

estimated by considering liquid foods namely milk, orange
and apple juice and varying the different process parameters,
the reduction in the microbial count is determined and
compared with control samples taken before PEF treatment.

During the application of oscillatory pulses at 20kV / cm

intensity to apple juice 27.5% increased percentage reduction
was achieved with the lower inductance of (10H) against
1.5mH. Nearly 52.8% increased reduction is evident for
higher (100) impulses against 50 impulses.

With orange juice m 28.03% higher reduction was achieved

when the inductance is 10H in contrast to 5mH thus
supporting the simulation results.

Contrary to the research reported that Bascillus subtilis are

resistant to PEF, in this study, when uniform field was applied
nearly 98.9% reduction is achieved during application of
50 kV/cm at 100 impulses. This might be due to the fact, that
the probability of Ec would have not been sufficient for
inactivation, whereas in this study 1.2/50 sec. pulse imparts
75.789 cal/ml energy sufficient to cause cell breakdown.

The Ec decreased with an increase in the dielectric constant.

For Milk of r = 69.5 required 60kV/cm for 38.52% of
microbial reduction but whereas orange juice r = 84 nearly
50.65% bacterial reduction was achieved for 20 kV/cm
supporting the simulation results

131

The results obtained during the PEF impact on milk increases

the feasibility of improving the preservation technology in
India which is fast becoming the number one dairy producing
nation in the world. Cows milk in India stays sterile for 4 to
6 hours if processed prior to normal preservation, definitely
the shelf life of the milk can be extended, thus equalizing the
demand countrywide.

An optimum field of 75kV / cm with a maximum of 100

impulses of 1.2 sec. rise time is required for effective
sterilization of E.coli. Pulses with short rise time (1.2 sec.)
results in 85.78% reduction of E.coli in milk against 32.52%
with 4.2 sec. pulse when 100 impulses at 75kV/cm is applied
due to the energy imparted by fast rising pulses.

Higher fat necessitates higher field and nearly 27.56% reduced

inactivation rate was achieved for 4.5% fat sample against
3.1% sample which is in par with simulation results.

The combined effect of irradiation (UV) and PEF treatment

proposed in this study seems to be a promising food
preservation method when compared to PEF alone. This is
evident from the experiments. Nearly 4.4% increased growth
reduction was achieved with combined effect of (UV) and
PEF in contrast to PEF alone still meeting the FDA
recommendation.

7.2.5

Digital colony count meter

The digital colony count meter using image processing technique

proposed here can be a boom for biochemists in reducing the ambiguities in

132

counting the colony forming units. Thus, diagnosis of serious diseases can be
made accurate, since this meter finds the exact number of bacterial colonies.

7.3

SCOPE FOR FUTURE WORK

Due to the advent of information technology in the present era, a

fully automated food processing unit can be developed, using the results
enlisted in this study. Further the hurdle of availability of samples for lab
scale treatment can be removed if fully closed loop controlled process is
employed.

Nutritive analysis can be done on the treated samples to ascertain

the advantages of PEF treatment. Since the proposed method of PEF with
irradiation gives promising results, this technique can be extended for other
liquid foods, still meeting the FDA recommendations. The electrical model
of the cell with pores can be used for calculating the TMP analytically for
effective electro permeabilization.

This study can be extended to mammalian cells, thus the growth of

cancerous cells can be mutated. The study reveals that electric field intensity
plays an important role in forming pores on the E. coli membrane. Here,
emphasis is made on irreversible breakdown however; these conclusions can
be effectively used in reversible breakdown that can be used for targeted drug
delivery. Further, E. coli can be made to think by varying Ec, which will
definitely pave a new arena in the field of bioprocessor, biosensors and even
as logical gates in nanotechnology fields.

Cost-effective analysis can be carried out for commercial outcome

of the process. Thus the future generation will be assured of safe processed

133

food resembling their freshly prepared counterparts with sensory attributes

preserved.

Thus this study on potential application of pulsed electric fields in

liquid food preservation will aid the food processing industry for improved
quality and safe processed food for future generation.