SJPM 1242 49
SJPM 1242 49
SJPM 1242 49
Yoghurt Production from Powdered Milk using Mixed Lactic Acid Bacteria
Starter Cultures
Obi CN*1, Olugbue VU2, Mpamugo CP1
1
Department of Microbiology, College of Natural Sciences, Michael Okpara University of Agriculture, Umudike, P.M.B.
7267 Umuahia, Abia State, Nigeria
2
Department of Science Laboratory Technology, Akanu Ibiam Federal Polytechnic, Unwana, P.M.B. 1007, Afikpo,
Ebonyi State, Nigeria
*Corresponding Author:
Obi CN
Abstract: Lactobacillus bulgaricus, Streptococcus thermophilus and Lactobacillus fermentum isolated from fermented
foods: yoghurt, “ugba” and “kunu-zaki” using MRS agar and a commercially acquired lactic acid bacteria (LAB) were
used to produce yoghurts samples A-H from Milksi Powered milk in a 5 hour fermentation process. The three LAB
isolates were used singly and in combinations as starter cultures. There was a maximum drop in pH (4-4.8) between the
second and third hour of fermentation while the yoghurt produced using S. thermophilus had the lowest pH (5.2) at the
temperature of 40oC. The optimum pH for the yoghurt production was 5.5 while the optimum temperature was 40 oC.
Yoghurt samples B and F had the highest moisture level (P<0.05) while Samples C, E and G had the highest dry matter
content. The highest ash content (0.8 ±0.01; P<0.05) was from sample F while Samples D and E had the highest crude
fibre content. Sample A had the highest crude fat value (3.95 a±0.01; P<0.05), Sample F the highest crude protein content
(3.95a±0.01; P<0.05) while the highest carbohydrate content was from Samples C, E, G and H (control). The control
Sample (H) had the highest and Hedonic test for sensory properties of the eight yoghurt samples showed that the eight
yoghurt samples were acceptable to the panelist indicating that the samples would compete favourably in the market with
commercially sold yoghurts.
Keywords: LAB, milk, optimum conditions, proximate analyses, yoghurts.
42
Obi CN et al.; Saudi J. Pathol. Microbiol.; Vol-1, Iss-2(Jul-Sep, 2016):42-49
isolation of LAB. The isolates were sub-cultured on
Although milk of various animals has been MRS Agar, subjected to microscopic, biochemical and
used for yoghurt production in various parts of the sugar fermentation tests and later stored in MRS Agar
world, most of the industrialized yoghurt production slant bottles and stored in the Refrigerator [10]. The
uses cow milk. Whole milk, partially skimmed milk, identification procedure given in Bergey’s manual of
skim milk or cream milk may be used. The milk used determinative Bacteriology [11] Cowan [12] were used
for yoghurt manufacture should be of the highest to characterized and identify the LAB isolates.
bacterial quality available and should be free from any
material that will impede or prevent the growth of the Production of youghurt
starter organism (antibiotics, preservative, disinfectant To each of eight glass beakers containing
and bacteriophages). 500ml of distilled water was added 200g of Milksi
powered milk and stirred continuously using a sterile
Fermentation of milk brings about many spatula till a homogenous solution was formed. The
changes in its chemical constituents, which have eight glass beakers were covered with aluminium foil
bearing on its nutritive value. Yoghurt has been found and then pasteurized and then cooled to 35oC and
to contain proteins, carbohydrate, fats, and high labeled A-H. Each of the LAB starter isolates was
percentage of lactic acid than other fermented milk inoculated in beakers A-C while a double combination
produce and it is rich in vitamin B complex. Yoghurt of the starter isolates were then inoculated in beakers D-
proteins are more digestible than that of fresh milk and F while beaker G had the three LAB isolates inoculated
that the partial hydrolysis of milk constituents in into it. A commercially procured LAB starter culture
yoghurt contributes to their increase digestibility. (control) was inoculated in beaker H and all the beakers
Cow’s milk is preferred for preparing yoghurt as having were incubated at 35oC for 6 hrs as adapted from Adam
low fat. This provides immunity; protects the consumer and Moss [1].
from cold, cough and strengthens body’s defense
mechanism as well as strengthens the collagen in the Determination of pH
skin. It lowers the blood pressure, bad cholesterol and The pH of the fermented milk samples
risk of heart attack. Yoghurt is one of the major sources (yoghurt) was measured with a pH meter with a glass
of vitamins and minerals and contains higher vitamins electrode every one hour.
(vitamins B12) than fresh milk.
Determination Of Titratable Acidity (T. A)
Recently, there is a growing interest to develop This was determined using the alkaline
a variety of fermented milk products for other beneficial titrimetric method of Sadler and Murphy [13]. 20ml of
purposes, particularly for health purposes and the sample was dispensed into conical flask and 3 drops
preventing of toxins produced by food- borne pathogens of phenolphthalein indicator was added. This was
and spoilage bacteria that enter human body [5-8]. The titrated against diluent standard alkaline solution (0.01N
beneficial effects of fermented milk products are NAOH solution). Titration was done until a persistent
produced by a variety of bioactive compound of lactic faint pink colouration was obtained. The total titratable
acid bacteria [9]. To this end, this work aimed at acidity was calculated using the formula below.
production of several yoghurt samples from powdered % Titratable acidity (T. A) =
milk using LAB isolates recovered from fermented
Where,
foods as well as determining the optimum conditions
T = Titre value
for the production of and the proximate analyses the
N = Normality of titrant
yoghurts.
W= Weight of sample used.
MATERIALS AND METHODS
Proximate Analysis
Sample collection
Proximate analysis of food is the determination
Samples of traditional fermented food:
of the major components of food. This includes
Yoghurt, ugba and Kunu-zaki were purchased from
moisture, fats (lipids), ash (mineral), protein
Ubani Market so also Milksi powdered milk samples
carbohydrate and fibre.
for the production of yoghurt and were taken to
Microbiology Laboratory, Michael Okpara University
1. Determination of Moisture Content
of Agriculture, Umudike for further analyses.
This was determined by the AOAC [14]
gravimetric method. A measured weight of the sample
Isolation of lactic acid bacteria (LAB).
(5g) was weighed into a previously weighed moisture
The various samples were serially diluted in
can. The sample in the can was evaporated to dryness
peptone water and 0.1ml of suitable aliquots was
over a steam bath and then dried in the oven 105 oCfor 3
streaked in duplicates onto De Man Rogosa Sharpe
hours in the first instance. It was cooled in a desicator
(MRS) Agar plates containing 50mg of Nystatin and
and weighed. It was then returned to the oven for
incubated at 37 C for 48 hours under anaerobic
further drying. Drying, cooling and weighing were
conditions using Anaerobic Gas Jar (Excello) for the
Cell arrangement
Gram reaction
Methyl Rred
morphology
Coagulase
Fructose
Catalase
Oxidase
Motility
Glucose
Maltose
Sucrose
Lactose
Colony
Nitrate
Urease
Isolate
Raised, + Rod in - - - + - - - + + - - + Lactobacillus
creamy with singles bulgaricus
whitish
colony
Raised, + Rod in - - - + + + - + + + + + Lactobacillus
circular, clusters fermentum
golden
yellow
colony
Raised, + Cocci - - - + - - - + - + + + Streptococcus
umbonate in thermophilus
colony clusters
Key
+= Positive
- = Negative
very
Dislike very
Neither like
Like slightly
moderately
moderately
extremely
attributes
Yoghurt
Sensory
nor like
slightly
sample
Dislike
Dislike
much
much
Total
Like
Like
Like
COLOUR A 0 2 2 1 0 0 0 0 5
B 0 3 2 0 0 0 0 0 5
C 3 1 1 0 0 0 0 0 5
D 1 2 2 0 0 0 0 0 5
E 4 1 0 0 0 0 0 0 5
F 2 1 2 0 0 0 0 0 5
G 0 2 3 0 0 0 0 0 5
H(control) 2 2 1 0 0 0 0 0 5
TEXTURE A 0 1 3 1 0 0 0 0 5
B 0 1 1 3 0 0 0 0 5
C 0 1 1 3 0 0 0 0 5
D 0 1 2 2 0 0 0 0 5
E 1 1 3 0 0 0 0 0 5
F 0 2 3 0 0 0 0 0 5
G 0 0 1 4 0 0 0 0 5
H(control) 3 2 0 0 0 0 0 0 5
FLAVOUR A 1 4 0 0 0 0 0 0 5
B 0 2 3 0 0 0 0 0 5
C 0 0 0 1 3 1 0 0 5
D 0 0 1 4 0 0 0 0 5
E 0 0 1 4 0 0 0 0 5
F 0 0 1 4 0 0 0 0 5
G 0 0 2 3 0 0 0 0 5
H(control) 0 2 2 1 0 0 0 0 5
TASTE A 0 0 1 2 2 0 0 0 5
B 0 0 0 2 1 2 0 0 5
C 0 0 0 1 1 3 0 0 5
D 0 0 0 2 1 2 0 0 5
E 0 0 2 3 0 0 0 0 5
F 0 0 2 2 1 0 0 0 5
G 0 1 3 1 0 0 0 0 5
H(control) 0 2 2 1 0 0 0 0 5
GENERAL A 0 0 2 3 0 0 0 0 5
ACCEPTABILITY B 0 0 3 2 0 0 0 0 5
C 0 3 2 0 0 0 0 0 5
D 0 2 3 0 0 0 0 0 5
E 4 1 0 0 0 0 0 0 5
F 1 4 0 0 0 0 0 0 5
G 0 0 2 1 1 0 0 0 5
H(control) 2 2 1 0 0 0 0 0 5