Available: https://doi.org/10.54117/ijamb.v4i1.

54 Research article

IPS Journal of Applied Microbiology and Biotechnology

IPS J Appl Microbiol Biotech, 4(1): 132-143 (2025)
DOI: https://doi.org/10.54117/ijamb.v4i1.54

Evaluating the Impact of Edible Coating on the Shelf Life of

Carrot (Daucus carota) and Green Pepper (Capsicum annuum)
Nnenna Jennifer Omorodion and Princess Uche Osuala

Department of Microbiology, University of Port Harcourt, PMB 5323 Rivers State, Nigeria.

*Corresponding author’s e-mail:

[email protected]; [email protected]

Abstract Article History

Edible coatings have emerged as a promising method to extend the shelf-life of fresh produce, including Received: 03 May 2025
carrots and green peppers. This study investigates the impacted influence of coating of the carrots and peppers Accepted: 11 May 2025
stored for 12 days. Standard microbial analysis was done using standard methods. The Total heterotrophic Published: 17 May 2025
bacteria count (THBC) in coated carrots and peppers ranged from 8.8x10⁴ CFU/g to 1.51x10⁶ CFU/g while Scan QR code to view
the THBC in pepper ranged from 4.4x10⁴ CFU/g to 9.4x10⁵ CFU/g. The THBC in uncoated carrot ranged
from 8.8x10⁴ CFU/g to 2.8x10⁶ CFU/g while the THBC in uncoated pepper ranged 9.4x10⁵ CFU/g to 1.9x10⁶
CFU/g. The Staphylococcus count in coated carrots ranged 6.9x10³ CFU/g to 9.4x10⁶ cfu/g while the
Staphylococcus count in pepper ranged from 4x10³ CFU/g to 8.8x10³ CFU/g. The Staphylococcus count in
Uncoated carrot and pepper ranged from 9.4x10³ CFU/g to 3.8x10⁴ CFU/g and 8.8x10³ CFU/g to 1.02x10⁴
CFU/g respectively. The fungi count in Uncoated carrot ranged 3x10³ CFU/g to 2.2x10⁴ CFU/g while the
fungi count in pepper ranged from 2.2x10³ CFU/g to 1.9x10⁴ CFU/g. The fungi count in coated carrots ranged
from 1.2x10³ CFU/g to 3x10³ CFU/g while the fungi count in pepper ranged from 1 4x10³ to 2.2x10³ CFU/g.
The Coliform count in coated carrots ranged from 2.2x10³ to 1.02x10⁴ CFU/g to while the Coliform count in
pepper ranged from 3.0x10³ CFU/g to 1.29x10⁴ CFU/g. The Coliform count in Uncoated carrot ranged from
7.6x10³ CFU/g to 1.12x10⁴ CFU/g while the Coliform pepper ranged from 5.9x10³ CFU/g to 1.42x10⁴ CFU/g. License: CC BY 4.0
Dominant bacteria included Staphylococcus aureus (24.6%), Bacillus subtilis (24.6%), and Escherichia coli
(14.7%), while fungi were predominantly Aspergillus niger (31%) and Aspergillus flavus (27.6%). Proximate Open Access article.
analysis revealed uncoated carrots had 5.40% ash, 30.40% moisture, and 42.62% carbohydrates initially, with
changes observed by day 15. Similarly, uncoated peppers had 80.32% moisture, 5.90% protein, and 5.50%
carbohydrates on day 0. Coated samples showed minimal changes in nutritional composition over time. The
study concludes that honey wax coating significantly reduces microbial load, extends shelf life, and maintains
the nutritional quality of carrots and peppers, recommending its application for preserving vegetables.

Keywords: Edible coating , Green peeper, Carrot, Microbial quality, Proximate and Vitamin composition
How to cite this paper: Omorodion, N. J., & Osuala, P. U. (2025). Evaluating the Impact of Edible Coating on the Shelf Life of Carrot (Daucus carota)
and Green Pepper (Capsicum annuum). IPS Journal of Applied Microbiology and Biotechnology, 4(1), 132–143. https://doi.org/10.54117/ijamb.v4i1.54.

Introduction According to Valero et al. (2013), a "edible coating" is a thin

The waxy layer that naturally covers the surface of all fruits covering of digestible material that protects food from oxygen,
and vegetables helps them retain water. Fruit respiration is external microorganisms, moisture, and solute movement. By
accelerated and the protective effect of the waxy covering is lowering respiration, gas exchange, oxidative reaction rates,
compromised when fruits or abrasions are wiped with moisture, and solute migration, among other physiological
wrapping paper. Therefore, another continuous or problems, edible coatings with a semi-permeable barrier seek
discontinuous film is applied to fruits and vegetables to to increase shelf life (Dhall, 2013; Shi et al., 2024).
provide a protective layer. Fruit and vegetable skins have also
inspired the use of edible coatings (Goldstein et al., 1992). Creating a barrier between respiration and transpiration is the
These are a thin covering of edible components that prevent primary purpose of an edible covering. As an antimicrobial
food soluble materials like oxygen and water from evaporating and antioxidant, it extends the shelf life and quality of both
(Bourtoom, 2008; Kotiyal and Singh, 2023). According to fresh and processed fruits and vegetables. Coatings can
McHugh and Krochta (1994), the coating is an essential enhance the appearance of green peppers and carrots and
component of the food that can be consumed as a whole. prolong their consumer attractiveness because they are staple
foods that are susceptible to microbial spoilage, which

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shortens their shelf life and causes financial losses a semi-permeable barrier on the surface of fruits and
(Giannakourou & Tsironi, 2021). vegetables, have been proposed as a potential solution to
extend freshness by reducing moisture loss, slowing
Fruits and vegetables experience the most postharvest loss respiration rates, and inhibiting microbial activity. However,
because of their perishable nature. Chopped fruits and while edible coatings have been explored for various fruits,
vegetables are growing in popularity as consumers choose their specific impact on the shelf-life and quality of vegetables,
fresh and organic items and as lifestyle changes are proven to particularly carrots and green peppers, remains under-
be successful. It is still challenging for the food business to researched. This study aims to evaluate how edible coating
continue producing cutting-edge food items for an extended affect the shelf-life and quality of carrot and green pepper,
period of time. providing insights into practical applications that can reduce
post-harvest losses and support food supply stability.
A balanced diet must include both fruits and vegetables (Riva Therefore, the aim of the study is to ascertain the impact of
et al., 2020). Biochemical changes such ethylene production, edible coating on the shelf life of two widely consumed
softening, pigmentation change, respiration rate, acidity, and commodities carrot and green pepper.
weight loss are necessary to reduce post-harvest losses.
Materials and Methods
One tactic to satisfy the present need is the use of edible films Samples Collection
and coatings (Tavassoli-Kafrani et al., 2016). Edible coatings A total of 24 samples of fresh carrots and green peppers of
preserve the quality of fruits and vegetables, which makes different sizes, varieties and the same maturity levels each was
them valuable for postharvest management strategies. Physical purchased from Choba Markrt in Port Harcourt, Rivers State.
alterations during preservation can be prevented by applying Using a sterile polythene bag properly labelled for each
an edible layer of proteins, polysaccharides, and lipids. The sample. For comparison, a set of fresh carrots and green
main function of an edible coating is to create a barrier peppers was treated with distilled water
between respiration and transpiration. Edible coatings act as
various functional compounds, including antioxidant and Preparation and Application of Edible Coating
antibacterial substances, enhancing the quality and extending Edible coating was prepared using natural polymer such as bee
the shelf life of fresh and minimally processed produce. These wax. The prepared edible coatings was applied evenly on the
coatings form a protective layer around fruits and vegetables, carrots and green peppers using dipping methods (Guilbert et
effectively covering the stomata. This action reduces the rate al., 1996).
of transpiration, subsequently decreasing weight loss (Prasad
et al., 2018). Their effectiveness has been shown in numerous Storage Conditions
fruits and vegetables, such as radish, potato, tomato, turnip, The coated and uncoated samples was stored under controlled
guava, plum, mango, apricot, banana, orange, and several conditions to stimulate real-world. The samples were placed
others. Natural biomolecules known as edible coatings are on a sterile tray for 12 days at ambient temperature and
utilized to enhance the appearance and preservation of fruits labelled as CC -For coated carrot sample, CP -For coated
and vegetables (Kotiyal et al., 2024). pepper sample, UCC -For uncoated carrot sample, UCP -For
uncoated pepper sample.
The safety and nutritional value of the fruits and vegetables are
improved by adding additional active substances to edible Microbial Analysis
films and coatings. To modify the structure and advance the Ten gram (10g) each of coated and uncoated carrots / green
characteristics of coatings, food grade extracts such as peppers were weighed into 90ml peptone solution for about 3
flavoring agents, pigments, antioxidants, and antimicrobials mins under sterile conditions and microbiological analysis was
can be added. This improves the quality of produce when carried out on day 0,3,6,9, and day 12. Ten (10) fold serial
applied to it (Nawab et al., 2017). Due to their edibility, non- dilution of the two samples (carrot and green pepper). A plastic
toxic makeup and cost-effectiveness in comparison to rack was arranged with ten (10) sterile test tubes containing
alternative synthetic coatings, edible coatings are in great 9ml of sterile 0.1 peptone water. Ten gram (10g) of each
demand from consumers today. The creation of edible sample was weighed using electronic weigh balance. The
biodegradable coatings and films as alternatives to synthetic weighed samples was carefully introduced into each of the
polymers, which cause less environmental harm than plastic conical flask containing 90ml peptone solution using a sterile
waste, has increased. spatula, vigorously mixed and labeled as dilution 10-1. A sterile
pipette was used to transfer 0.1 ml solution from dilution 10 -1
A lot of research is currently being done on edible coating and to the next test tube (dilution 10-2). Subsequent stepwise
the materials used in the edible coating in food processing transfers was carried out using a sterile pipette for each transfer
technology to decrease waste and limit loss and is healthy, until dilution 10-4 is reached. The content of each test tubes
functional for storage and market distribution. The shelf-life of was shaken vigorously before transferring an aliquot (0.1 ml)
fresh produce like carrots and green peppers is often limited of the mixture into the next test tube.
by rapid spoilage, primarily due to water loss, microbial
growth, and enzymatic degradation. This rapid deterioration Enumeration of Microorganisms
leads to significant food waste and financial losses in supply Total Heterotrophic Bacteria Count (THBC): From the
chains and households, posing a challenge for maintaining diluted samples, 0.1ml aliquots was plated on Plate count agar
food security and sustainability. Edible coatings, which create (PCA). The spread method was employed, and inoculation was

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performed aseptically using sterile hockey sticks. Plates were was conducted for further identification and confirmation of
labelled according to their dilutions. Total heterotrophic the pure bacterial isolates which include:
bacteria count (THBC) of the coated and uncoated carrots as
determined by culturing the diluted samples on plates count Isolation and identification fungi isolates
agar plates using pour plate method. The culture plates was Fungi colonies will be isolated and streaked on freshly
incubated for 24 hours at 37 0C. After incubation the colonies prepared PDA plates incubated for 3-5days at 37 0C. After 3-5
was manually counted and described according to their days pure colonies will be stored in freshly prepared PDA
morphological characteristics and results was recorded in slants for further microscopic identification. The colour,
CFU/g. shape, edge, size and texture of the fungi isolates will be
microscopically identified and results will be recorded.
Total Coliform Count (TCC): From the diluted samples,
0.1ml aliquots was plated on MacConkey agar (MAC). The A sterile inoculating needle were used to pick discrete colonies
spread method was employed, and inoculation was performed on the fungal plates and streaked on freshly prepared PDA
aseptically using sterile hockey sticks. Plates were labelled plates. The inoculated plates were incubated at room
according to their dilutions. Total coliform count (TCC) of the temperature 37oC for 3-5 days to obtain pure isolates. The
coated and uncoated carrots was determined by culturing the fungal isolates from coated carrots and green peppers was
samples on MacConkey agar plates using pour plate method. identified microscopically by the observation of their mycelia
The culture plates was incubated for 24 hours at 370 C. After under the microscope at ×40 objectives with lactophenol
incubation the colonies was manually counted and described cotton blue stain and results would be recorded.
according to their morphological characteristics and results
was recorded in CFU/g. Physiochemical Analysis
The proximate composition of the samples was analyzed for
Total Staph Count (TSC): From the diluted samples, 0.1ml moisture, ash, protein, fat and fibre by the method of AOAC
aliquots was plated on Mannitol salt agar (MSA). The spread (2005). The carbohydrate content was calculated by
method was employed, and inoculation was performed subtraction method Pomeranz (1971), Jabeen et al. (2019)
aseptically using sterile hockey sticks. Plates were labelled .Standard method of AOAC (2012) was used to determine
according to their dilutions. Total Staphylococcal count (TSC) Vitamins.
of the coated and uncoated carrots was determined by culturing
the samples on Mannitol Salt agar plates using pour plate Statistical analyses
method. The culture plates was incubated for 24 hours at 37 0 Analysis of variance (ANOVA) was used to compare means at
C. After incubation the colonies was manually counted and p < 0.05. This analyses was performed to visualize the
described according to their morphological characteristics and association between the microbial loads of the coated and
results was recorded in CFU/g. uncoated samples using SPSS (Statistical Package for the
Social Sciences), also known as IBM SPSS Statistics, is a
Total Fungi Count (TFC): From the diluted samples, 0.1ml software package used for the analysis of statistical data.
aliquots was plated on Potato dextrose agar (PDA). The spread
method was employed, and inoculation was performed Results
aseptically using sterile hockey sticks. Plates were labelled Total heterotrophic Bacteria Count of Coated and
according to their dilutions. Total Fungi count (TFC) of the Uncoated Carrots and Pepper Studied
coated and uncoated carrots was determined by culturing the The total heterotrophic bacteria count of the coated and
samples on plates dextrose agar plates using pour plate uncoated carrot and pepper showed varying counts. On day 0,
method. The culture plates was incubated for 3- 5 days at 370 uncoated carrot had a count of 8.8x104 CFU/g while the
C. After incubation, the colonies will be manually counted. uncoated pepper sample had a THBC of 9.4x105 CFU/g. On
After incubation the colonies was manually counted and day 3, the coated carrot had counts of 9.6x104 CFU/g while
described according to their morphological characteristics and coated pepper had a 4.4x104 CFU/g count; the uncoated carrot
results was recorded in CFU/g. had a count of 2.8x105 CFU/g while the uncoated pepper had
a count of 1.0x105 CFU/g. The final day of analysis (Day 15)
The number of bacteria colonies was determined for each was not analyzed due to excessive deterioration (spoilage) but
plates. The total heterotrophic bacterial count was obtained day 12 showed higher counts as the coated carrot had a
from plate count agar culture after 24 hours of incubation, the 1.16x105CFU/g count while the coated pepper had a count of
total coliform count was obtained from MacConkey agar 3.0x105 CFU/g. The uncoated carrot had 6.5x106 CFU/g while
culture after 24 hours of incubation, and the total the uncoated pepper had a count of 3.1x106 CFU/g. Details of
staphylococcus spp count was obtained from mannitol salt counts obtained on days 6 and 9 are as presented in Figures 1
agar culture after 24 hours of incubation. The colony counts below
were expressed as colony forming units per gram (CFU/g).
Individual bacteria colonies were selected from the previously Staphylococcus Counts of Coated and Uncoated Carrots
incubated media based on their morphological features such as and Pepper Studied
colony size, color, texture, elevation and odor. These were then The staphylococcus count of the coated and uncoated carrot
sub-cultured and streaked on freshly prepared nutrient agar and pepper showed varying counts. On day 0, uncoated carrot
plates using the streak plate method and incubated at 37 0C for had a count of 9.4 ×104 CFU/g while the uncoated pepper
24 hours to obtain pure colonies. Various biochemical tests sample had a staphylococcus count of 8.8×103 CFU/g. On day

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3, the coated carrot had counts of 7.2x103 CFU/g while coated showed higher counts as the coated carrot had a 6.9x103 CFU/g
pepper had a 6.9x103 CFU/g count. The uncoated carrot had a count while the coated pepper had a count of 4.0x103 CFU/g.
count of 9.4x103 CFU/g while the uncoated pepper had a count The uncoated carrot had 2.5x104 CFU/g while the uncoated
of 8.8x103 CFU/g. The final day of analysis (Day 15) was not pepper had a count of 2.0x104 CFU/g. Details of counts
analyzed due to excessive deterioration (spoilage) but day 12 obtained on days 6 and 9are as presented in Figures 2 below.

8
Count (Log CFU/g)

4
Coated carrot
2
Uncoated carrot
0
Day 0 Day 3 Day 6 Day 9 Day 12
Dasys Monitored

Figure 1: Bar chart of total heterotrophic Bacteria Count of Coated and Uncoated Carrots

8
Count (Log CFU/g)

6
4
Coated pepper
2
Uncoated pepper
0
Day 0 Day 3 Day 6 Day 9 Day 12
Days monitored

Figure 2: Bar chart of total heterotrophic Bacteria Count of Coated and Uncoated Pepper

6
Count (Log CFU/g)

5
4
3
2 Coated carrot
1 Uncoated carrot
0
Day 0 Day 3 Day 6 Day 9 Day 12
Days monitored

Figure 3: Bar chart of Staphylococcus Counts of Coated and Uncoated Carrots Studied

4.4
Counts (Log CFU/g)

4.2
4
3.8
Coated pepper
3.6
3.4 Uncoated pepper
3.2
Day 0 Day 3 Day 6 Day 9 Day 12
Days monitored

Figure 4: Bar chart of Staphylococcus Counts of Coated and Uncoated Pepper Studies.

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Total Coliform Counts of Coated and Uncoated Carrots Fungi Counts of Coated and Uncoated Carrots and
and Pepper Studied Pepper Studied
The total coliform count of the coated and uncoated carrot and The total fungi count of the coated and uncoated carrot and
pepper showed varying counts. On day 0, uncoated carrot had pepper showed varying counts. On day 0, uncoated carrot had
a count of 1.02x104 CFU/g while the uncoated pepper sample a count of 3.0x103 CFU/g while the uncoated pepper sample
had a coliform count of 1.29x104 CFU/g. On day 3, the coated had a fungi count of 2.2x103 CFU/g. On day 3, the coated
carrot had counts of 5.6x103 CFU/g while coated pepper had a carrot had counts of 1.6x103 CFU/g while coated pepper had a
4.7x103 CFU/g count. The uncoated carrot had a count of 1.4x103 CFU/g count. The uncoated carrot had a count of
1.12x104 CFU/g while the uncoated pepper had a count of 3.4x103 CFU/g while the uncoated pepper had a count of
1.42x104 CFU/g. The final day of analysis (Day 15) was not 3.9x105 CFU/g. The final day of analysis (Day 15) was not
analyzed due to excessive deterioration (spoilage) but day 12 analyzed due to excessive deterioration (spoilage) but day 12
showed higher counts as the coated carrot had a 2.2x103 CFU/g showed higher counts as the coated carrot had a 1.2x103 CFU/g
count while the coated pepper had a count of 3.1x103 CFU/g. count while the coated pepper had a count of 1.8x103 CFU/g.
The uncoated carrot had 7.6x103 CFU/g while the uncoated The uncoated carrot had 2.2x104 CFU/g while the uncoated
pepper had a count of 5.9x103 CFU/g. Details of counts pepper had a count of 1.9x104 CFU/g. Details of counts
obtained on days 6 and 9are as presented in Figures 5 below. obtained on days 6 and 9are as presented in Figures 6 below
.
5
Counts (Log CFU/g

2 Coated carrot

1 Uncoated carrot

0
Day 0 Day 3 Day 6 Day 9 Day 12
Days monitored

Figure 5: Bar chart of Total Coliform Counts of Coated and Uncoated Carrot Studied
4.4
Counts (Log CFU/g)

4.2
4
3.8
Coated pepper
3.6
3.4 Uncoated pepper
3.2
Day 0 Day 3 Day 6 Day 9 Day 12
Days Monitored

Figure 6: Bar chart of Total Coliform Counts of Coated and Uncoated Pepper Studie
5
Count (Log CFU/g)

4
3
2 Coated carrot
1 Uncoated carrot
0
Day 0 Day 3 Day 6 Day 9 Day 12
Days Monitored

Figure 7: Bar chart of Fungi Counts of Coated and Uncoated Carrot Studied

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5
Count (Log CFU/g)
4

3
Coated pepper
2
Uncoated pepper
1

0
Day 0 Day 3 Day 6 Day 9 Day 12
Days Monitored

Figure 8: Bar chart of Fungi Counts of Coated and Uncoated Pepper Studied
8
Mean Count (Log Cfu/g)

0
Coated Carrot Uncoated carrot Coated Pepper uncoater pepper
Sample

Figure 9: Bar Chart of the Mean Total Heterotrophic Bacteria Count of Coated and Uncoated Green Pepper and Carrot
4.1
Mean Count (log CFU/g)

4.05
4
3.95
3.9
3.85
3.8
3.75
Coated Carrot Uncoated carrot Coated Pepper uncoater pepper
Sample

Figure 10: Bar Chart of the Mean Total Staphylococcus Count of Coated and Uncoated Green Pepper and Carrot
4.3
Mean Counts (Log CFU/g)

4.2
4.1
4
3.9
3.8
3.7
3.6
3.5
3.4
Coated Carrot Uncoated carrot Coated Pepper uncoater pepper
Sample

Figure 11: Bar Chart of the Mean Total Coliform Count of Coated and Uncoated Green Pepper and Carrot

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Mean Counts (log CFu/g)

5
4
3
2
1
0
Coated Carrot Uncoated carrot Coated Pepper uncoater pepper
Sample

Figure 12: Bar Chart of the Mean Total Fungi Count of Coated and Uncoated Green Pepper and Carrot

Table 1: Frequency of Occurrence of Bacteria Isolated from Coated and Uncoated Carrots and Pepper Studied
Carrot Pepper
Bacteria genera Coated Uncoated Coated Uncoated Total
n (%) n (%) n (%) n (%) N (%)
Escherichia coli 0(0.0) 5(29.4) 0(0.0) 4(19.0) 9(14.7)
Staphylococcus spp. 5(35.7) 4(23.5) 3(33.3) 3(14.3) 15(24.6)
Bacillus spp. 4(28.5) 1(5.9) 3(33.3) 7(33.3) 15(24.6)
Micrococcus spp. 5(35.7) 4(23.5) 3(33.3) 2(9.5) 14(22.9)
Serratia mascercens 0(0.0) 3(17.6) 0(0.0) 0(0.0) 3(4.9)
Enterobacter spp. 0(0.0) 0(0.0) 0(0.0) 5(23.8) 5(8.2)
Total 14(22.9) 17(27.9) 9(14.7) 21(34.4) 61(100.0)

Table 2: Frequency of Occurrence of Fungi Isolated from Coated and Uncoated Carrots and Pepper Studied
Carrot Pepper
Bacteria genera Coated Uncoated Coated Uncoated Total
n (%) n (%) n (%) n (%) N (%)
Aspergillus niger 4(100.0) 3(23.1) 2(50.0) 0(0.0) 9(31.0)
Aspergillus flavus 0(0.0) 4(30.7) 1(25.0) 3(37.5) 8(27.6)
Mucor spp 0(0.0) 2(15.4) 1(25.0) 1(12.5) 4(13.8)
Alternaria spp. 0(0.0) 2(15.4) 0(0.0) 2(25.0) 4(13.8)
Saccharomyces cerevisiae 0(0.0) 2(15.4) 0(0.0) 2(25.0) 4(13.8)
Total 4(13.8) 13(44.8) 4(13.8) 8(27.9) 29(100.0)

Proximate composition of Coated and Uncoated Carrots values for uncoated carrot for Ash, Moisture content, Lipid,
and Pepper Studied crude protein, crude fibre and carbohydrates were: 5.5%,
The proximate composition of coated and uncoated carrot and 26.2%, 0.63%, 5.90%, 16.14% and 44.8% while that of
pepper samples studied showed that one Day 0, Ash content of Uncoated pepper were 7.50%, 50.32%, 0.64%, 0.40%, 11.2%,
the respective samples (Uncoated carrot and pepper) studied and 33.75%. The values of coated carrots for Ash, Moisture
were 5.40 and 5.0%; Moisture content were 30.40% and content, Lipid, crude protein, crude fibre and carbohydrates
80.32%; Lipid content were 0.53 and 0.64%; Crude protein were: 5.40%, 26.60%, 0.64%, 5.90%, 16.14% and 44.8%
3.96% and 5.90%,. Crude fibre were 16.40% and 5.50% and while that of the coated pepper samples were 7.50%, 51.11%,
Carbohydrate 42.62% and 5.50% respectively. All samples 0.61%, 0.40%, 11.2% and 33.85%. (See Table 3).
showed varying figure from day 0 figures on day 15. The

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Table 3: Proximate composition of Coated and Uncoated Carrot and Pepper Studied on days 1 and 9.
Sample Ash Moisture Lipid Crude Crude Carbohyd
Code (%) Content (%) Protein Fibre rate (%)
(%) (%) (%)
Day 0
Uncoated 5.40 30.40 0.53 3.96 16.40 42.62
Carrot
Uncoated 5.0 80.32 0.64 5.90 3.0 5.50
Pepper
Day 9
Uncoated 5.5 26.2 0.63 5.90 16.14 44.8
Carrot
Coated 5.40 26.60 0.64 5.90 16.14 44.8
Carrot
Uncoated 7.50 50.32 0.64 0.40 11.2 33.75
Pepper
Coated 7.50 51.11 0.61 0.40 11.2 33.85
Pepper

Table 4: Vitamin composition of coated and uncoated carrot and green pepper
Sample Vitamin A (ug) Vitamin B (ug)

Coated carrot 846.19 0.36

Uncoated carrot 1758.42 0.82

Coated pepper 113.96 14.31

Uncoated pepper 178.45 23.76

Discussion counts obtained from this fresh produce both coated and
Microbial quality of coated and uncoated carrot and green uncoated are as follows
pepper
The impact of honey wax coating on the shelf life of green The Total Heterotrophic Bacteria Count (THBC) revealed that
peppers and carrots was examined in this study. Honey wax, honey wax coating significantly reduced bacterial counts in
an edible coating, is used to enhance the appearance of food both carrots and peppers (p < 0.05) as shown in fig .1 and fig
and ensure its safety due to its environmentally beneficial .2. On day 0, the uncoated carrot had a THBC of 8.8 x 104
properties. During handling, processing, and storage, they CFU/g, while the uncoated pepper had a notably higher THBC
serve as a barrier to keep out gasses and moisture. Through of 9.4 x 105 CFU/g. By day 3, coated carrots had a THBC of
their action or the addition of an antimicrobial chemical, it 9.6 x 10⁴ CFU/g, while coated peppers had a lower count of
improves food safety and lessens food deterioration. The study 4.4 x 10⁵ CFU/g. In contrast, uncoated samples continued to
evaluated honey wax's efficacy as a natural antimicrobial show elevated bacterial growth, with uncoated carrots
coating in prolonging shelf life by comparing the bacteria reaching 2.8 x 10⁴ CFU/g and uncoated peppers 1.0 x 10⁵
counts in coated and untreated samples. The antibacterial CFU/g. (p < 0.05). On day 12, coated carrots had a THBC of
effectiveness of honey wax in prolonging shelf life was 1.16 x 10⁵ CFU/g and coated peppers 3.0 x 10⁵ CFU/g, while
evaluated by measuring the microbiological quality of coated uncoated samples exhibited excessive spoilage, with counts as
and untreated samples during a 12-day period. Prior research high as 6.5 x 10⁴ CFU/g for carrots and 3.1 x 10⁶ CFU/g for
has emphasized the advantages of coatings made of honey and peppers. Comparing counts with the regulatory guidelines.US
wax, pointing to their antibacterial qualities and capacity to Food and Drug Administration states less than 105 cfu/g,
form a barrier that inhibits moisture loss and respiration, European Union states less than or equal to 106 cfu/g and
maintaining freshness. Yaman and Bayoindiri (2002) discuss International Organization for standardization states less than
how cherries are affected by edible covering. According to 105 cfu/g for fresh fruit and vegetables, these guidelines are
their research, this coating may help cherries lose weight while line with this present study.
also improving their firmness, ascorbic acid concentration,
titratable acidity, and skin color. Additionally, it extended the Staphylococcus counts also exhibited a marked decrease in
cherries' shelf life. According to Lee et al. (2003) and Bai et coated samples (p < 0.05) as shown in fig .3 and fig .4.
al. (2003), who investigated the effect of edible coating on Initially, uncoated carrots had 9.4 x 10⁴ CFU/g, whereas
apple slices and varieties, these coatings also demonstrated uncoated peppers had a lower count of 8.8 x 10³ CFU/g. By
their ability to lengthen the shelf life of apples. Microbial day 3, coated carrots had a staphylococcal count of 7.2 x 10³
CFU/g, while coated peppers had 6.9 x 10³ CFU/g, reflecting

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a reduction compared to uncoated samples (9.4 x 10³ CFU/g 2018). In a study on effect of agar agar based coating on
for carrots and 8.8 x 10³ CFU/g for peppers). On day 12, the minimally processed cloves it was observed that filamentous
coated carrot samples exhibited staphylococcal counts of 6.9 x fungus and aerobic measophilic were inhibited and the coating
103 CFU/g, while coated peppers showed 4.0 x 10³ CFU/g, resulted in reduction of respiration rate of clove Geraldine et
both significantly lower than uncoated samples (p < 0.05). al., (2008).
According to the International Commission of Microbiological
Specification for fruit and vegetables (ICMSF, 1994 1998) Impact of mango edible film on fresh mango quality and shelf
states Total staphylococcus count less than 10 4 cfu/g and life during storage was evaluated by Sothornvit and
Food and Agriculture Organisation of the United Nation states Rodsamran (2008) this showed that film provided good
less than to 105 cfu/g. is acceptable. This guideline oxygen barrier properties and had enough mechanical strength.
corroborates with this study. The coating’s efficacy in It also prolonged the shelf life of the mangoes. The impact of
reducing Staphylococcus spp. aligns with previous findings modified atmosphere packaging and honey dip treatment as
that honey’s osmotic effect and hydrogen peroxide release can edible coating on minimally processed grapes was evaluated
hinder Gram-positive bacteria, including Staphylococcus by Sabir Ali et al., (2011). Their study stated that it reduced
(Shakila et al., 2016). weight loss of berries. It also maintained the quality of
minimally processed grapes by retarding quality loss and berry
Coliform counts further illustrate honey wax’s effect on decay. Result showed that honey dip edible coating act as
microbial inhibition as shown in fig 5 and .6. On day 0, barrier to moisture and resist to water vapour diffusion during
uncoated carrots had 1.02 x 10⁴ CFU/g, while uncoated the cold storage and gave good adherence to berry surface and
peppers had 1.29 x 10⁴ CFU/g. By day 3, coated carrots had also increase shelf-life.
reduced coliform counts of 5.6 x 10³ CFU/g, and coated
peppers had 4.7 x 10³ CFU/g. On day 12, the coated carrots Effect of coating based on dewaxed shellac and Aloe vera on
exhibited 2.2 x 10³ CFU/g, and coated peppers had a slightly tomato fruit was evaluated by Chauhan et al., (2015). Their
higher count of 3.1 x 10³ CFU/g, in contrast to uncoated study stated that they contain optimum barrier properties
samples, which displayed elevated spoilage. These are slightly towards O₂, CO₂ and water vapour and they effectively
higher compared to the findings reported by Pilon et al. (2006). prevented anaerobiosis. It also prolonged the shelf life of
According to the International commission of microbiological tomatoes. According to Sogvar et al., (2016) studied the
specification for food (ICMSF, 1994,1998) reported that impact of edible coating such as aloe vera with ascorbic acid
coliform of less than 102 CFU/g is considered acceptable in on strawberries. Their study suggested that coating reduced
fruit and vegetables and Food and Agriculture organization weight loss and inhibiting the growth of mesophilic yeast and
(FAO) of the united nation less than 103cfu/g comparing the molds. This coating enhanced the shelf life of strawberries.
count with ICMSF, (1994 .1998), The findings in this study is These studies is line with this present study which showed that
slightly higher than the acceptable limit. Honey wax coating’s organisms counts were greater for the coated than with non-
impact on coliform counts supports findings by Sapper & coated i.e. control towards the end of storage. The antibacterial
Chiralt, (2018), which highlighted that edible coating activity of the coating and the barrier produced around the cut
antimicrobial properties can control coliform growth, thereby fruit surface account for the decrease in coated samples.
enhancing food safety. Differences between carrots and Microbial activity is suppressed by the use of edible coatings
peppers in coliform counts could relate to the nutrient on fruits.In this study Honey’s acidity, hydrogen peroxide
composition, with peppers providing a more conducive content, and phenolic compounds are key contributors to its
environment for coliform bacteria due to higher moisture and antimicrobial action. The wax component further slows
sugar levels. Omorodion & Oge (2022). microbial access to nutrients by reducing moisture exchange
and oxygen permeability, effectively creating an anaerobic
These results are consistent with those of Zhang et al. (2004), environment that inhibits bacterial growth (Becerril-Sánchez
Tanada Palmu et al. (2005), Maftoonazad et al. (2005), Osae et al., 2021)
(2014), and Nosiba et al. (2018), who all showed a decrease in
the microbial count on fresh produce, indicating that the The morphological and biochemical analysis identified six
natural antimicrobial qualities of honey wax (edible coating) genera of bacteria Staphylococcus aureus, Micrococcus luteus,
successfully inhibited bacterial growth. But in a research by Bacillus subtilis, Serratia merscenscens, Escherichia coli, and
Salihah et al. (2023), wax-coated tomatoes showed an even Enterobacter aerogenes present in both coated and uncoated
higher decrease in microbial count. Variations in coating samples. Staphylococcus and Micrococcus, frequently found
composition and application techniques may be the cause of in fresh produce, were the most prevalent genera, occurring in
this discrepancy. Due in significant part to its acidity and both types of samples across all days. These bacteria is similar
hydrogen peroxide release, honey's antibacterial properties to that isolated by Omorodion and Oge, (2022). Similar
may work better on produce with a rougher surface, such studies have documented that bacteria such as Bacillus and
peppers (Aziz & Karboune, 2018). Additionally, as high- Escherichia coli are commonly isolated from raw vegetables
moisture conditions tend to support bacterial development and are strongly influenced by the application of protective
more readily, variations in the vegetable's inherent nutritional coatings. For instance, Numes et al. (2023) observed that
and moisture profiles may also lead to varying bacterial growth edible coatings reduce bacterial populations by creating a
rates. Additionally, each vegetable has a naturally rough physical barrier that reduces moisture loss and limits bacterial
surface; peppers, with their higher water content, may offer adherence. In comparing the bacterial load between carrots and
bacteria a more favourable environment (Aziz and Karboune, peppers, the study found higher counts in uncoated samples,

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likely due to increased surface exposure and nutrient showing significant fungal proliferation. According to ICMSF
availability Siriwardana and Wijesekara (2021). and FDA guidelines recommend that fungi count of less than
105 CFU/g is considered acceptable in fruit and vegetables.
Lipid coatings (Bee wax) are good barriers to moisture loss. In The effectiveness of honey in reducing fungal growth is
addition to preventing water loss, lipid coatings have been corroborated by studies that emphasize honey’s antifungal
used to reduce respiration, thereby extending shelf life, and to action due to its osmotic pressure and low water activity
improve appearance by generating a shiny product in fruits and (Mahmud et al., (2015). Differences in fungal counts between
vegetables. Coatings that include lipid solids up to 75% can be carrots and peppers may result from differences in their surface
used to improve coating performance without diminishing structures and moisture content, which affect fungal adherence
moisture-barrier properties (Martin-Polo et al., 1992) The and growth rates .Omorodion and Oge (2022).
presence of Enterobacter and Serratia spp., primarily in
uncoated samples, suggests that the honey wax coating was The study identified Mucor sp., Aspergillus flavus, Aspergillus
effective in reducing these potential spoilage bacteria, a niger, Saccharomyces cerevisiae, and Alternaria spp. as the
finding consistent with Raybaudi-Massilia et al. (2008), who predominant fungi present in both coated and uncoated
observed lower spoilage bacteria levels on wax-coated samples. As shown in table 2, Aspergillus niger (31%) and
produce. Aspergillus flavus (27.6%) were the most frequent. The
reduced occurrence of these fungi in coated samples confirms
The frequency of bacterial genera as seen in table 1, showed the antifungal efficacy of honey wax, as documented in studies
Staphylococcus and Bacillus spp. as the most prevalent in both where honey compounds inhibit fungal spore formation Yash
coated and uncoated samples, with Staphylococcus accounting Pandya et al. (2023). The frequent presence of Aspergillus
for 24.6% and Bacillus 24.6% of isolates. Coated samples, species, which are common post-harvest fungi, suggests a need
however, had notably reduced frequencies, indicating the for fungal control in produce handling. The honey wax coating
coating’s effectiveness in curbing bacterial presence. likely limited spore germination by reducing oxygen
Comparable study have shown that wax and honey coatings availability and moisture, both critical factors for fungal
inhibit bacteria like Staphylococcus by acting as a barrier to proliferation (Baldwin et al., 2019). Interestingly, Aspergillus
environmental oxygen, which limits bacterial respiration and niger showed the highest occurrence (31%) among uncoated
growth. Tajik and Jalali (2009). samples, especially in Green pepper.

The presence of Escherichia coli, Staphylococcus aureus, The presence of Mucor sp., Aspergillus flavus, Aspergillus
Micrococcus luteus, Bacillus subtilis, and Serratia marcescens niger, Saccharomyces cerevisiae, and Alternaria sp. in carrots
in carrots and green peppers has significant public health and green peppers has significant public health implications.
implications. These microorganisms can serve as indicators of Mucor sp. can cause mucormycosis in immunocompromised
poor hygiene practices during cultivation, harvesting, storage, individuals. Saccharomyces cerevisiae, though used in
or handling. Their potential health risks range from foodborne fermentation, can occasionally cause infections in vulnerable
illnesses to opportunistic infections. Escherichia coli, individuals. Alternaria spp. produce mycotoxins that may lead
especially pathogenic strains, poses a severe risk of to allergic reactions and respiratory issues. Aspergillus flavus
gastrointestinal illnesses, including diarrhea and hemolytic produces aflatoxins, potent carcinogens linked to liver cancer,
uremic syndrome, if contaminated produce is consumed raw while Aspergillus niger can cause opportunistic infections and
(Malabadi et al., 2024). Its presence often indicates fecal spoilage (Pavon et al., 2012). It was observed that edible
contamination, highlighting inadequate sanitation. coating positively affected shelf life and microbial load
Staphylococcus aureus, known for producing enterotoxins, in fresh cut fruits. Shelf life of all fresh-cut fruits was
can cause food poisoning characterized by nausea, vomiting, significantly enhanced by edible coating.
and abdominal cramps. Improper storage and temperature
abuse can enhance toxin production. Micrococcus luteus, Physicochemical composition of coated and uncoated
although generally non-pathogenic, can be opportunistic, carrot and green pepper
causing infections in immunocompromised individuals. The physicochemical analysis on day 0 showed that the
Bacillus subtilis is a spore-forming bacterium that can survive uncoated carrot had an ash content of 5.40%, moisture content
harsh conditions. Though typically harmless, it can cause of 30.4%, lipid content of 0.53%, crude protein of 3.96%,
spoilage and foodborne illness through toxin production. crude fiber of 16.4%, and carbohydrate content of 42.62%.
Serratia marcescens, can cause infections, especially in This is lower compared to the findings reported by Omorodion
hospital environments, and may pose risks if transferred from and Oge, (2022). In contrast, uncoated peppers had higher
contaminated produce (Caggiano et al., 2021). moisture content (80.32%). The lower carbohydrate content
(5.5%), reflecting the distinct composition of each vegetable.
The fungal count demonstrated that honey wax coating was By day 12, moisture in uncoated carrots dropped to 26.2%,
also effective against fungal growth as shown in fig 7 and fig while uncoated peppers reduced to 50.32%, indicating
8. Initially, uncoated carrots had a fungal count of 3.0 x 10 3 significant dehydration in uncoated samples. This is slightly
CFU/g, and uncoated peppers had a count of 2.2 x 10³ CFU/g. similar to the findings reported by Omorodion and Oge,
By day 3, coated carrots had a reduced fungal count of 1.6 x (2022). The coated samples, however, retained higher
10³ CFU/g, and coated peppers had 1.4 x 10³ CFU/g. On day moisture levels (26.6% in carrots and 51.11% in peppers),
12, coated carrots had a fungal count of 1.2 x 10³ CFU/g, while likely due to the honey wax layer reducing water loss. The
coated peppers had 1.8 x 10³ CFU/g, with uncoated samples stability in ash, crude protein, and carbohydrate content further

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highlights the coating’s preservation effect, as observed in 10.1080/10408398.2016.1194256. Epub 2017 Jun 12. PMID:
this study on honey-coated produce. 27437876.
Bai J, Robert HD, Elizabeth A, & Baldwin (2003). Coating selection for
delicious and other apples. Postharvest Biology and Tech. 28:381-
The vitamin composition of coated and uncoated carrot and 390.
green pepper samples highlights the impact of coating on Bai J, Victorine A, Robert, H.D, James. MP, Elizabeth A,M &
nutrient retention. The uncoated carrot had higher Baldwin.(2003) Formulation of zein coating for apples (Malus
concentrations of Vitamin A (1758.42 µg) and Vitamin B domestica Borkh). Postharvest Bio and Tech. 28:259-268.
(0.82 µg) compared to the coated carrot (846.19 µg and 0.36 Baldwin, E. A., Nisperos-Carriedo, M. O., & Hagenmaier, R. D. (2019).
Edible coatings to preserve quality and safety of fresh and minimally
µg, respectively). Similarly, uncoated green pepper exhibited processed fruits and vegetables. Postharvest Biology and
higher Vitamin A (178.45 µg) and Vitamin B (23.76 µg) levels Technology, 38(1), 127-132
compared to the coated sample (113.96 µg and 14.31 µg, Becerril-Sánchez AL, Quintero-Salazar B, Dublán-García O, & Escalona-
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retention or measurement of these vitamins, potentially due to Relationship with Antioxidant Activity, Botanical Origin, and Color.
interactions with the coating material or a barrier effect Antioxidants (Basel). 10(11):1700.
Bourtoom T. 2008. Review Article: Edible films and coatings:
reducing nutrient exposure. characteristics and properties. Int. Food Res. J. 153: 237-248
Caggiano, G., Triggiano, F., Diella, G., Apollonio, F., Lopuzzo, M.,
Conclusion Mosca, A. & Montagna, M. T. (2021). A possible outbreak by
This study highlights the efficacy of honey wax coatings in Serratia marcescens: genetic relatedness between clinical and
extending the shelf life and enhancing the microbial safety of environmental strains. International journal of environmental
research and public health, 18(18), 9814.
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