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Microbiological analysis of raw milk, pasteurized milk and yogurt samples

collected from different areas of Dhaka city, Bangladesh

Article  in  Journal of Bangladesh Academy of Sciences · June 2015

DOI: 10.3329/jbas.v39i1.23655

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Journal of Bangladesh Academy of Sciences, Vol. 39, No. 1, 31-36, 2015

MICROBIOLOGICAL ANALYSIS OF RAW MILK, PASTEURIZED MILK

AND YOGURT SAMPLES COLLECTED FROM DIFFERENT AREAS OF
DHAKA CITY, BANGLADESH

TASMINA RAHMAN*, TANZIA AKON, IFTIKHARUN NESSA SHEULI AND

NAIMA HOQUE

Department of Microbiology, Stamford University Bangladesh, Dhaka-1217, Bangladesh

ABSTRACT

A total of 300 different milk and yogurt samples were collected from Dhaka city and analyzed
for total viable bacterial count (TVBC), total coliform count (TCC) and presence of several
pathogens such as Escherichia coli, Salmonella spp., Shigella spp., Vibrio spp. and Listeria
monocytogenes. Raw milk samples collected from Moghbazar area showed highest TVBC (4.2 ×
106 cfu/ml) and samples from Mohammadpur area showed lowest count (3.5 × 10 3 cfu/ml). All the
pasteurized milk samples showed TVBC of 1.9 × 102 to 2.8 × 103 cfu/ml, and TCC only in one
sample Yogurt samples (open) collected from vendor showed TVBC of 9.1 × 103 to 8.2 × 107
cfu/ml, and TCC only in sample collected from Dhanmondi, Dhaka. The packed yogurt samples
showed TVBC of 2.2 × 102 to 6.1 × 103 cfu/ml, and TCC only in one sample. All the milk and
yogurt samples were found contaminated with E. coli and Shigella-like species. Listeria
monocytogenes was not detected in any of the samples studied.

Key words: Raw milk, Pasteurized milk, Yogurt, Listeria monocytogenes, Coliform

INTRODUCTION

Milk is considered to be an ideal food and supposed to contain proteins, fats,

carbohydrates, vitamins and mineral salts. For this rich nutritional composition, bacteria
can easily multiply in milk (Park et al. 2007). The consumption of milk and milk
products in Bangladesh is very low compared to neighboring countries (BER 2007),
however, several reports document the presence of food borne pathogens, like
Escherichia spp., Listeria spp., Staphylococcus spp. and other pathogenic bacteria in the
udder of the animal, collected milk, untreated milk and milk samples becomes threat to
human health (Baird-Parker and Tompkin 2000, de Buyser et al. 2001). These data reflect
the unsanitary condition of the collection centers and the negligence of hygienic rules
during the milking and carrying of the raw milk as well. According to Bangladesh
Standard and Testing Institute (BSTI), the pasteurized milk sample must not contain
TVBC more than 2 × 104 cfu/ml. The guideline also allows the presence of less than 10
cfu/ml of coliform bacteria and 0 cfu/ml of fecal coliforms (BSTI 2002, Jay 2003).

*
Corresponding author: <[email protected]>.
32 RAHMAN et al.

Another popular and demanding nutritious food is cultured milk or yogurt. It is a

fermented food with preserve nutrient value in the milk with a characteristic fresh taste
derived from the action of bacteria on all or part of the lactose to produce several organic
acids (Tamine and Robinson 2004). If produces from untreated or contaminated milk
samples, yogurt can pose some health hazard and are labile to contamination.

There are mainly three sources of contamination of milk and milk products: inside
and outside of udder, from the surface of handling and storage equipments. Post
pasteurization contamination has received most of the attention and is considered to be a
factor, which limits shelf life in the majority of cases (Tekinsen et al. 2007). In
Bangladesh, BSTI (The Bangladesh Standards and Testing Institution) compels some
chemical and sanitary requirements for pasteurized milk and milk cream; however, no
guidelines are available for UHT milk (Hassan et al. 2009). Although there is little milk
pockets specially milk-vita, and some other established dairy farms where surplus milk is
readily available in Bangladesh, this perishable product has never received particular
attention in hygienic distribution to the consumers (Khan et al. 2008).

The objectives of the present study were to assess the bacteriological quality of milk and
several milk product samples giving priority to detect the presence of several pathogenic
bacteria like E. coli, Salmonella, Shigella-like species, Vibrio and Listeria monocytogenes.

MATERIALS AND METHODS

Twenty raw milk and 20 vendor’s yogurt samples were collected from each of five
selected area (Moghbazar, Siddeswari, Lalmatia, Dhanmondi and Mohammadpur) of
Dhaka Metropolitan city (total of 200 samples). These areas have been selected as they
are densely populated and several schools and colleges have been located around.
Furthermore, 10 samples each of pasteurized milk and packed yogurt were also collected
from each of 5 commercially available brands. Samples were chosen randomly from
different super shops and local markets. Raw samples were collected in sterile flask,
however, packed milk and yogurt samples were purchased as sealed pack by checking the
valid expiry dates. Samples were transferred to the laboratory for quality testing within
30 minutes it was collected.

The samples were serially diluted up to 10-7. Standard plate count (SPC) method
recommended for dairy products (APHA 1960) was followed for quantitative analysis of
bacteria.

A volume of 10 ml raw sample was inoculated in buffered peptone broth, selenite

cystine broth, alkaline peptone broth for enrichment of Escherichia coli, Salmonella spp.,
Shigella spp. and Vibrio spp., respectively and incubated for 6 hours. After that, 0.1 ml of
enriched sample was spread over MacConkey agar, Salmonella-Shigella agar and
QUALITY ANALYSIS OF MILK AND MILK PRODUCTS 33

Thiosulfate citrate bile Salt sucrose (TCBS) agar media and incubated at 37C. On the
otherhand, 0.1 ml raw sample was spread over Listeria identification media and incubated
at 37C. Following incubation for 24 hours, typical colonies for each genus were
considered as positive growth and microscopic observation of the isolated bacteria was
done by Gram staining. Confirmation of the organisms was performed by different
biochemical tests, such as Triple Sugar Iron (TSI) agar, catalase test, citrate utilization
test, indole test, motility test, methyl red test, Voges Proskaur test and urease production
test (Cappuccino and Sherman 1996).

RESULTS AND DISCUSSION

This study tested a total of 200 open vendor milk and milk products, 100 raw milk
and 100 open yogurt, to assess microbiological quality of those milk products. The
laboratory investigation confirms presence of coliform and other bacteria in milk
samples. The count of tested milk products are presented in Table 1. The count of TVBC
and coliform bacteria in raw milk were higher than the acceptable limit set by Bangladesh
Standard Testing Institute (BSTI 2002). Previously published study in Sylhet Agricultural
University area in Bangladesh (Ara et al. 2010) also found the presence of high number
of total viable count and total coliform count in the raw milk of samples. This was found
that in raw milk the average TVBC was highest in the samples collected from Moghbazar
area and the coliform count was highest in the samples collected from Siddeswari area.

Table 1. Total bacterial counts (cfu/ml) and coliform counts (cfu/ml) in raw milk and open
vendor yogurt samples.

Raw milk (n = 100) Open yogurt (n = 100)

Collection area Average TVBC Average TCC Average TVBC Average TCC
(cfu/ml) (cfu/ml) (cfu/ml) (cfu/ml)
Moghbazar 4.2  106 3.4  103 8.2  107 0
Siddeswari 5.2  10 5
4.1  10 3
3.1  10 6
0
Mohammadpur 3.5  10 3
2.5  10 1
9.1  10 3
0
Lalmatia 4.7  10 4
3.8  10 1
4.0  10 4
0
Dhanmondi 3.6  10 3
5.1  10 1
7.3  10 5
2.8  103

As yogurt is a cultured food, presence of bacterial population is obvious and there is

no specific acceptable limit of bacterial population for yogurt is available. However,
presence of pathogenic bacteria or coliforms is not acceptable and presence of coliform is
often used as a parameter of proper sanitary condition in different countries (Tamine and
Robinson 2004). The entire open vendor yogurt samples showed a large number of
heterotropic bacterial growth (Table 1). Highest growth was found in the sample
collected from Moghbazar area but the sample collected from Dhanmondi area showed
presence of coliform which was a critical point of concern. It is assumed that the
34 RAHMAN et al.

competition and low pH acts as an inhibitory factor for bacterial growth. Similar result
was reported that in available market curd in different countries like in Bangladesh
(Chowdhury et al. 2011), Sudan (Haj et al. 2007) and Lahore (Khalid et al. 2008).

In this study, the authors identified bacteria in pasteurized milk as well, but the
average TVBC in pasteurized milk was less than threshold level of BSTI. They detected
coliform in only one commercial pasteurized milk brand sample, brand three, which is
not acceptable as per BSTI guidelines (Table 2). Rizwan et al. (2011) observed the
presence on Bacillus brevis in pasteurized milk sample and suggested the post
pasteurization process. Choudhury (2008) also reported the presence of coliform in
pasteurized milk samples. There can be several steps in pasteurized milk production like
proper temperature maintaining, handling and post pasteurization processing that might
introduce bacteria in milk.

Table 2. Total bacterial counts (cfu/ml) and coliform counts (cfu/ml) in pasteurized
milk and packed yogurt samples.

Pasteurized milk (n = 50) Packed yogurt (n = 50)

Brand Average TVBC Average TCC Average TVBC Average TCC
(cfu/ml) (cfu/ml) (cfu/ml) (cfu/ml)
1 2.5  102 0 4.3  102 0
2 2.4  10 2
0 2.2  10 2
0
3 2.8  10 3
1.4  10 1
3.2  10 3
0
4 6.1  10 2
0 5.2  10 2
0
5 1.9  10 2
0 6.1  10 3
0

The commercially packed yogurt samples were found to harbor low range of
heterotrophic bacteria with the highest number of 3.2 × 103 cfu/ml without coliform
bacteria.

Raw milk samples collected from five different areas were found to be contaminated
with several pathogenic bacteria like Escherichia coli, Salmonella spp., Shigella species
and Vibrio spp. (Table 3). Presence of any of these bacteria in food samples is not
acceptable as they can cause food borne illness (Rahman et al. 2008). Similar data were
collected from the open vendor yogurt samples. This reflected the poor sanitary measure
and under processed food parameters. The scenario is better in case of pasteurized milk
and packed yogurt sample. However, presence of E. coli and Shigella-like species
indicated the lower quality food samples. A study of quality assessment of milk product
pointed to the presence of E. coli and Listeria monocytogenes in tested samples (Cordano
and Rocourt 2001). A fact of hope that, no sample was found to be contaminated with L.
QUALITY ANALYSIS OF MILK AND MILK PRODUCTS 35

monocytogenes. This organism is a potential cause of food poisoning found in variety of

milk and dairy products (Farber 2000).

Table 3. Presence of pathogenic bacteria in different milk samples.

Number of samples contaminated with pathogenic bacteria

Bacteria Raw milk Pasteurized milk Vendor yogurt Packed yogurt
(n = 100) (n = 50) (n = 100) (n = 50)
E. coli 52 5 15 3
Salmonella spp. 6 0 13 0
Shigella-like species 11 2 6 2
Vibrio spp. 1 0 1 0
Listeria spp. 0 0 0 0

CONCLUSION

The laboratory investigation confirms that most of the raw milk and open vendor
yogurt samples that have been tested were contaminated with huge load of heterotropic
bacteria. Bacterial contamination of these products assumed to be originated from poor
hygiene sense of the handler and improper processing, preservation and marketing
strategy. These products usually are found to be sold in an open condition with less
sanitary measure which can introduce contaminants into these products. On the contrary,
the pasteurized milk and packed yogurt are sold in sealed packs and in sufficient sanitary
conditions. These physical barriers are useful to prevent microbial contamination.
However, few products exceeded the acceptable limit of bacteria which indicate the
inadequate processing and packaging of these goods. Therefore, proper guidance and
education for the processor, handler and consumer are necessary to maintain a consistent
healthy life of consumer body.

ACKNOWLEDGEMENT

The authors thank the authorities of the Stamford University Bangladesh for
providing laboratory facilities and financial support.

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(Received revised manuscript on 1 March,, 2015)

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