FACULTY OF FISHERY AND FOOD SCIENCE

STM 3110

ADVANCED FOOD MICROBIOLOGY (K2)

LAB 2 : MICROBIOLOGICAL QUALITY IN SPICES

LECTURER’S NAME : DR. NURMAHANI BINTI MOHD MAIDIN

Group : 4D

NO NAME MATRIC NUMBER

1 NUR AZIANI BINTI MOHD MARSIDI S60111

2 NOR SYAMSINA NADHILAH BINTI AHMAD NIZAR S59114

3 LAU CAI LING S58425

METHODOLOGY

Equipments

Weighing balance, stomacher, sterile stomacher bag, incubator, micropipettes, Bunsen burner,
colony counter, and Petri dish

Materials

Peptone water, spice, PCA agar, DRBC agar, and TGE agar

Serial Dilution and Incubation of PCA, DRBC and TGE agar

Aerobic and anaerobic incubation
RESULT

Table 1.0: Plate count of PCA (37°C, 24 Hours)

Dilution 10-3 10-4 10-5

Colonies per
plate

Too numerous to 26 (Est.) 9 (Est.)

count, TNTC (>300)

Explanation TNTC 26 TFTC

(Rejected as it is (Accepted as it is (Rejected as it is

more than 250) in the range of less than 25)
25-250)

CFU/g 26 × (1/10-4) × (1/1) = 2.6×104

Table 2.0: Plate count of DRBC (Room Temperature, 5 Days)

Dilution 10-2 10-3 10-4

Colonies per
plate

1 (Est.) 0 (Est.) 0 (Est.)

Explanation - - -

CFU/g -

Table 3.0: Plate count of TGE (55°C, 48 Hours)

Dilution 10-2 10-3 10-4

Colonies per
plate

52 (Est.) 10 (Est.) 7 (Est.)

 Explanation 52 TFTC TFTC

(Accepted as it is (Rejected as it is (Rejected as it is

in the range of less than 25) less than 25)
25-250)

CFU/g 52 × (1/10-2) × (1/1) = 5.2×102

Table 4.0: Plate count of Mesophilic Aerobe (37°C, 24 Hours)

Dilution 10-3 10-4 10-5

Colonies per
plate

1 (Est.) 1 (Est.) 1(Est.)

Explanation - - -

CFU/g -
Table 5.0: Plate Count of Anaerobic sporeformers in the CO2 chamber (37°C, 48 hours)

Dilution 10-2 10-3 10-4

Colonies per
plate

1 (Est.) 1 (Est.) 0 (Est.)

Explanation - - -

CFU/g -

*TNTC = Too Numerous to Count (>250)

*TFTC = Too Few to Count (<25)

DISCUSSION

Spices and herbs are natural items with different flavors, colors, and aromas that are widely
used in culinary preparation. They are natural taste enhancers and ideal low-salt ready meals
options (Mitchell et al., 2013). Basically, all the spices and herbs are planted in temperate and
tropical areas. Warm temperatures and high humidity are ideal for the growth of
microorganisms, particularly some that are harmful (Garbowska, et al., 2015). Microbiological
quality, namely the load of total heterotrophs or Enterobacteriaceae, is frequently used as an
indicator of a region's hygienic situation where spices are grown and processed. Spices, like
many other agricultural commodities, are subjected to a wide spectrum of microbial
contamination in the environment during collection, processing, and retail markets due to dust
and waste water. Depending on the ultimate application, contaminated spices may cause a
microbiological concern. Spices may be harmful to people's health because they are frequently
added to foods that do not go through any further processing or are consumed raw.

In the laboratory class, our group has selected curry powder spices to find out the
microbiological quality in it. At the beginning we were instructed to make a series of dilutions for
the usage in PCA, DBRC and TGE agar, also for aerobic and anaerobic incubation experiments.
−1
To make a 10 dilution: we mixed 10 gram of curry powder with 90 ml of peptone water. Serial
−5
dilution has been made up to 10 . Next, after incubating the Principal component analysis
(PCA) plates for 24 hours at 37℃, we could see the number of colonies per plate for each
−3
dilution. Firstly, on the plate with 10 dilution factor, there are too many colonies to be counted,
we consider it more than 300 counts, so based on the formula we rejected as it is more than
−4
250. Moreover, for the dilution of 10 . We counted 26 and we accepted it as it is in the range of
−5
25 to 250 of colonies. Meanwhile, for the dilution of 10 , there are estimately 9 colonies
observed on the plates, we rejected due to less than 25 colonies. To compare with the other
spices; coriander powder, cumin and red pepper,the number of colonies that appeared was
approximately equal to the number of colonies present in the curry powder sample as Mally
curry powder.
 Next, we used the Dichloran-rose Bengal (DRBC) agar in order to identify whether yeast and
mold are present in the food products since it has been proposed as a general purpose medium
−2
for fungal enumeration. We found only at dilution of 10 , there was fungal growth, but no fungal
−3 −4
growth at dilution of 10 and 10 which estimately zero of the colony. Because of that, this
curry powder product may be safe to use in cooking or food production. Furthermore, the plate
count of Tryptone Glucose Extract (TGE) agar is used for microbe cultivation and enumeration
in dairy and bottled water products. Over all three plates, we found some resulting colony
growth. The lowest dilution factor was found to have many colonies growing compared to the
−2 −3 −4
high dilution factor. The number of colonies on the dilution factors of 10 , 10 and 10 is
−2
estimated to be 52, 10 and 7 respectively. As per results showed, for dilution factor 10 It was
accepted as it is in the range of 25-250 colonies counted while for the other dilution factors we
did reject it as it is less than 25 colonies counted.

We also ran the experiments of plate count of Mesophilic aerobe for 24 hours at 37°C and
plate count of anaerobic sporeformers in the CO2 chamber at 37°C,for 48 hours. Different
dilution factors were needed for these two experiments, for instance in Mesophilic aerobic
−3 −4 −5
experiment we used dilution factor of 10 , 10 and 10 while for anaerobic sporeformers
−2 −3 −4
10 , 10 and 10 . We observed that no number of colonies could be calculated due to no
colony growth on the plates. We conclude, there was a low count of microbial load in the curry
powder sample. Thus, spices in our country are safe and sanitary levels are high in the food
industry production.

Malaysia is widely known as a food lover’s heaven due to the diversity of our cultures and
foods that consist of an ample amount of spices varying for each cuisine. According to new
research by Helgi Library, Malaysia is ranked as the 12th country with the most spice
consumption in the world, just behind Nepal (Which Country Eats the Most Spice?, 2019). From
the result of this experiment, it is safe to say that the spices available in our country are
unquestionably safe for consumption as it has a very low colony count. The major reason
behind this occurrence is because of the high sanitary level in the factories that produce and
manufacture spice products. In Malaysia, food manufacturers must take great care of their
hygiene level during food processing to acquire the certification that complies with the standard
and requirements from the Ministry of Health (MOH) such as Hazard Analysis and Critical
Control Point (HACCP), Good Manufacturing Practice (GMP), and Makanan Selamat
Tanggungjawab Industri (MeSTI) certificate (HACCP, MESTI, GMP Certification in Malaysia |
Food Safety Guidelines, 2021). The purpose behind these certificates is to ensure the safety of
food products manufactured before it reaches the consumers hence decreasing the chances
and risk of getting sick from foodborne illnesses such as food poisoning. The experiment's
findings indicate that the production of spices and herbs in Malaysia is conducted under
hygienic circumstances.

CONCLUSION

In conclusion, the muller curry result indicated good hygienic condition. In this lab, the
colonies selected were between 25 to 250 in one plate because the colonies are single
colonies. We learnt to use the pour plate technique, conducting in different agar. The plate count
in the different agar has their function, which PCA is to determine the quality of food and DRBC
is used to enumerate viable yeasts and molds of food product and TGE is used for the
cultivation and enumeration of microorganisms commonly in dairy products. There is absence in
enumeration of mesophilic aerobic and anaerobic spore formers in the miller curry, meaning that
there is no spore forming. We also learnt an aseptic technique which is important in the process
of isolating culture to prevent any contamination of spices from pathogenic microorganisms.
Serial dilution is also vital in preparing the single and visible colony for study to determine the
microbial count easily.
REFERENCE

1. Freitas. R, Nero, L.A & Carvalho, A.F. (2009). Technical note: Enumeration of mesophilic

aerobe in milk: Evaluation of standard official protocols and Perrifilm aerobic count

plates. https://www.sciencedirect.com/science/article/pii/S002203020970623X

2. Garbowska, M., Berthold-Pluta, A. & Stasiak-Rozanska, L. (August, 2015).

Microbiological quality of selected spices and herbs including the presence of

Cronobacter spp. https://www.sciencedirect.com/science/article/pii/S0740002015000088

3. HACCP, MESTI, GMP Certification in Malaysia | Food Safety Guidelines. (2021,

December 28). Mandreel.

https://www.mandreel.com/malaysia/haccp-gmp-mesti-certification/

4. Mitchell, M., Brutnon, N. P., Fitzgerald, R. J., & Wilkinson, M. G. (2013). The Use of

Herbs, Spices, and Whey Proteins as Natural Flavor Enhancers and Their Effect on the

Sensory Acceptability of Reduced-Salt Chilled Ready-Meals. Journal of Culinary Science

& Technology, 11(3), 222–240. https://doi.org/10.1080/15428052.2013.769869

5. Which Country Eats the Most Spice? (2019, June 28). Helgi Library.

https://www.helgilibrary.com/charts/which-country-eats-the-most-spice/