Bacterial contamination of fruits and vegetables:

 Frequent contacts with farm soil, irrigation waters, insects, animals, or humans during the
growing or harvesting period, and in the processing plants.
 Improperly processing and/or re-contamination with human bacterial pathogens during
handling.
 No post-harvest processing intervention step effective enough to eliminate these pathogens.

Factor affecting Bacterial Attachment to fruits/Vegetables surfaces:

 Nature of surfaces (apples and tomatoes have smooth natural waxy fruit surfaces).
 Surface irregularities (crevices and pits in the netting).
 Surface structure and physiological characteristics of bacteria
 Physical properties of any substratum, in this case the fruit surfaces.
 Cell surface charge
 Hydrophobicity
 Presence of microbial surface appendages such as flagella and fimbriae (pili) as
 Extracellular polysaccharides

Physical and chemical treatments are often used in food processing to eliminate or at least reduce the
presence of pathogenic and spoilage microorganisms in foods.

The use of chlorinated water (usually up to 200 ppm) as a post-harvest washing treatment.

1–5% hydrogen peroxide solutions

There is much interest from the produce industry in developing more processing intervention
technologies that can kill pathogenic bacteria on fruit surfaces without damaging quality and or
contributing to physical changes on produce surfaces or in the produce itself. In this study, we proposed
using combinations of different organic acid mixtures as antimicrobials to substitute for chlorine-based
sanitizers for inactivation of bacterial pathogens on fruit surfaces. The organic acids were chosen based
on our earlier studies. Therefore, the objectives of this research were to: 1) investigate the strength of
attachment of Salmonella on whole apple and tomato surfaces and 2) study the efficacy of a combined
organic acid antimicrobials developed in our laboratory in reducing attached bacteria and finally
compare the efficacy against known and often used 200 ppm chlorinated and hydrogen peroxide
washing treatments on the same produce types.

ll inoculations were done by dipping 5 apples and 10 tomatoes at a time in

separate sterilized plastic bowls containing 1 L of bacterial inoculum described
above with constant rotation for 5 min until all produce were inoculated.
Inoculated produce was placed in a biosafety cabinet to air-dry for 2 h at room
temperature (22 ± 1 °C). After drying, the pathogen-inoculated apples and the
tomatoes were washed with sterilized tap water immediately, 30 min, 60 min,
120 min, 360 min, 540 min and 1440 min post inoculation duration as
described below. Also, the efficacy of antimicrobial treatments for reducting
populations of aerobic mesophilic bacteria and artificially
inoculated Salmonella was investigated.

Table 2: Surviving Salmonella bacteria on whole apples and tomatoes at 20 min post
inoculation and log reduction after antimicrobial treatments.

Produce Treatments Surviving populations (log Log reduction (log

CFU/g) CFU/g)
Apple Control 4.6 ± 0.18a -----b
Water 3.8 ± 0.14 b 0.8 ± 0.2c
H2O2 (1.5%) 1.0 ± 0.12c 3.6 ± 0.12a
Chlorine-200 ppm 1.8 ± 0.10c 2.8 ± 0.12 b
Organic acid 0.6 ± 0.2 d 4.0 ± 0.10a
solutiond
Tomatoesc Control 3.4 ± 0.14a ----b
Water 2.8 ± 0.14 b 0.6 ± 0.12 d
H2O2 (1.5%) 0.3 ± 0.02c 3.0 ± 0.10c
Chlorine-200 ppm 0.9 ± 0.03c 2.5 ± 0.12c
Organic acid NDd 3.4 ± 0.10a
solutiond

Table 1: Initial population of aerobic mesophilic bacteria and population

of Salmonella bacteria inoculated on whole apples and tomatoes before and after
antimicrobial treatments.

Produce Treatments Aerobic mesophilic bacteria (log Salmonella (log

CFU/g) CFU/g)
Apple Control 4.5 ± 0.12a 4.6 ± 0.12a
Water 4.2 ± 0.12a 3.8 ± 0.14 b
H2O2 (1.5%) 2.7 ± 0.12 b 1.0 ± 0.12c
Chlorine-200 ppm 2.9 ± 0.10 b 1.8 ± 0.10c
Organic acid 1.8 ± 0.02c 0.6 ± 0.2 d
solutionc
Tomatoesb Control 3.1 ± 0.13a 3.4 ± 0.12a
Water 2.8 ± 0.12a 2.7 ± 0.12a
H2O2 (1.5%) 1.6 ± 0.02 b 0.3 ± 0.02 b
Chlorine-200 ppm 1.9 ± 0.03 b 0.6 ± 0.03 b
Organic acid 1.0 ± 0.03c NDb
solutionc
The produce wash with water did not result in significant (p > 0.05) changes
in the surviving populations of attached Salmonella on apple surfaces
throughout the treatment time.
At 30 min post inoculation and treatment with H2O2 and chlorine, the
surviving populations on apple surfaces were reduced and averaged 2 logs for
duration of up to 120 min post treatment;
however, these populations were slightly reduced to below 2 logs at 360 min
post treatment and beyond that time.
The surviving populations on apples treated with H2O2 and chlorine were not
significantly (p > 0.05) different, until at 540 min and 1440 min after
treatments. However, the wash with organic acid solution led to significant
(p < 0.05) reductions compared to H2O2 and chlorine treatments.
The same survival trend and log reduction was observed on tomato surfaces
treated at similar time intervals and again, the new combined organic acid
solution reduced more Salmonella populations than the H2O2 and chlorine
treatments

Strength of salmonella attachment (SR-value) on apples and tomatoes surfaces

The result showed an increased in SR-values from 30 min to 360 min post
inoculations
the SR-values after 360 min post treatment remained stable with little change.
This observation is consistent with the log reduction numbers observed at
360 min post washing treatments.
The average initial SR-value determined on apples and grape tomatoes within
120 min post inoculation and washing treatments averaged 0.22 and 0.25,
respectively and significantly increased to 0.8 at 1440 post inoculation and
treatment for both produce. A slightly higher SR-value for Salmonella bacteria
was recorded on tomatoes at 120 min and 1440 min post inoculation and
washing treatment suggesting that any pathogen attached on apple and
tomato surfaces for more than 120 min before decontamination would be
difficult to detach or inactivate with chemical antimicrobial treatments.
Although significant populations of Salmonella bacteria were recovered on
apple surfaces than on tomato, the attachment strength was similar, and the
SR-values were not significantly (p > 0.05) different.
A similar increase in the strength of attachment on both apples and tomatoes
was observed, suggesting that the efficacy of washing treatments at < 2 h post
inoculation would have an improved efficacy, but would not provide accurate
understanding of bacteria reduction or inactivation on produce surfaces. The
populations of Salmonella bacteria on apples treated with 200 ppm chlorine,
1.5% hydrogen peroxide and the combination of organic acid solution
immediately following inoculations were higher than when antimicrobial
treatments were applied on produce at time period ≥120 min post inoculation
and stored. These log reductions were not significantly (p > 0.05) different
when treatment was applied at 30 min post inoculation on each produce.
However, changes in population reductions were observed when antimicrobial
treatments were applied above 120 min post inoculation and exposure. The
antimicrobial activity of the combined organic acid solution showed consistent
microbial reduction above 120 min and was bactericidal on aerobic mesophilic
bacteria and on Salmonella bacteria populations that were already inoculated
on apples and tomatoes. The antimicrobial efficacies of the organic acid
solution and their subsequent effects on reduction of Salmonella populations
were significantly (p < 0.05) higher than that of 1.5% hydrogen peroxide and
200 ppm chlorine treatments.