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EFFECT OF MICROBIAL STARTER CULTURE ON SOME MICROBIAL PROPERTIES OF FERMENTED SAUSAGE

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Abdulrahman & Qoja (2020): Effect of microbial starter culture on fermented suasage Nov 2020 Vol. 23 Issue 18
Annals of Tropical Medicine & Public Health http://doi.org/10.36295/ASRO.2020.231817
EFFECT OF MICROBIAL STARTER CULTURE ON SOME MICROBIAL
PROPERTIES OF FERMENTED SAUSAGE
RanjbarShwanAbdulrahman1*, Abduljabar Omer Qoja2
1,2Department of Food Technology, College of Agricultural Engineering Sciences, Salahaddin University-
Erbil, Kurdistan Region, Iraq
*Corresponding author E-mail: Ranjbar.abdulrahman@su.edu.krd (Abdulrahman)
ABSTRACT
The study was aimed to investigate the effect ofmicrobial starter culture on some microbial
properties of fermented sausage,themixed starter culture which was used were composed from Lactobacillus
curvatus, Pediococcusacidi lactici and Staphylococcus xylosus after study microbiological analysis of
sausages and counting total viable count, lactic acid bacteria (LAB) of sausages and sausages yeasts and
molds counts (log CFU/g )at fermentation period (0,5,10)day and after stored period for (30,60,90) days for
control and starter inoculated sausages were the obtained result the total viable count increased respectively
from zero time to end day of productionwhich was5.44 to 8(log CFU/g ) for control and 5.88 to 8.37(log
CFU/g ) for starter inoculated sausages after these period at the stored period the number decreased. The
Lactic acid bacteria (LAB) countingdone and the rangeincreased respectively from zero time to 10thday of
production was 4.67 to 8.13 (log CFU/g ) for control and 6.11 to 8.7 (log CFU/g ) for starter inoculated
sausages after these period at the stored period the range of LAB decreased however the yeast and mould
counting undertaken for both type of sausagesand theywas lowered from first day till end day of storage from
zero time to endday of stored was 5.32 to 3.78 (log CFU/g ) for control and 5.28 to 3 (log CFU/g )for starter
inoculated sausages.
Keywords: starter culture, total viable count, lactic acid bacteria (LAB) of sausages, sausages yeasts and
molds counts
How to cite this article: Abdulrahman RS, Qoja AO (2020): Effects of microbial starter culture on some microbial properties of
fermented sausage, Ann Trop Med & Public Health; 23(S18): SP231817. DOI: http://doi.org/10.36295/ASRO.2020.231817
INTRODUCTION
Fermented sausage is prepared from seasoned, crude meat that's stuffed in casings and
is allowed to ferment and mature (1, 2). Inoculation of the sausage is a batter with a starter culture composed
of chosen lactic acid bacteria (LAB), i.e. homofermentative lactobacilli and/or pediococci, and Gram-
positive, catalase- positive cocci (GCC), i.e. nonpathogeni, coagulase-negative staphylococci, improves the
quality and safety of the final product and standardizes the production process (1-3).Various microorganisms at
the environment and raw material are source of microorganisms; naturally contaminate dry sausage mixtures
(4). Among them, lactic acid microorganisms (LAB) are responsible for the most event during dry sausage
Abdulrahman & Qoja (2020): Effect of microbial starter culture on fermented suasage Nov 2020 Vol. 23 Issue 18
Annals of Tropical Medicine & Public Health http://doi.org/10.36295/ASRO.2020.231817
ripening the fermentation of carbohydrates. Can lead to the improvement of normal flavours and tastes
through by lactate oxidation, proteolysis, degradation of amino acids and lipolysis (5).
Fermentation is a worldwide and ancient preservation technique, possibly one of the
oldest strategies recognized (6). It is generally employed to keep or enhance the organoleptic attributes and
microbiological safety of foods. Indigenous microorganismshave been responsible for fermentation
traditionally, however starter cultures can now be brought to set off fermentation and favorable
processing conditions can be chosen to make sure desired best quality (1,7) These techniquesencourage the
improvement of a suitable safe microflora, which is vital for preventing the outgrowth of
spoilage microorganism and food-borne pathogens (8).The major function of lactic acid bacteria is to convert
fermentable sugars in the sausage batter to lactic acid, thereby contributing to product safety via developing
unfavorable conditions for pathogens and spoilage organisms. (9, 10).However Staphylococcus and Kocuria
(formerly Micrococcus) are the main representatives of this family. These micro-organisms have important
enzymes like some proteases and lipases that contribute to flavor generation. (11).Hydrolysis, the partial
degradation of the lipid fraction in fermented sausages, is one of the most important biochemical processes,
which takes place as a end result of the action of glycerol ester hydrolases (12-15).
MATERIALS AND METHODS
Sausage Preparation And Fermentation Procedure
The sausages were produced with a simple modification according to (16-18) lean beef (80% w/w) and
animal internal tissue fat (20% w/w). Meat and animal internal tissue fat were minced and mixed with
sausage Spices obtained from local market added bay range (3.5 g/100 g), salt (2 g/100 g), raw garlic paste
(0.2 g/100 g), and crystal sugar (0.3 g/100 g). Half of the obtained mixture was inoculated with Safe Pro F-
LC starter culture which for each 25-gram packet ofF-LC starter culture is sufficient for 100 kg of meat about
10 cfu/g; mixed culture, Safe Pro F-LC starter, Chr. Hansen, Germany starter cultures Lactobacillus
curvatus, Pediococcusacidi lactic and Staphylococcus xylosus in a convenient freeze-dried And another half
use as a control without starter.The mixture stuffed in natural beef casings the ripening was performed as
follows in a traditional method for 10 days (control: C and starter inoculated: S). The conditions natural
conventional maturation room the primary arrange comprised of 2 days drying with the relative humidity
(RH) of 85% and a temperature of 22 0CAirspeed velocity (m/s) 0.1-0.2 that was at that point diminished to
15-17ºC were the temperature lowered after that 5 days to (normal 7.11 ± 3.20°C)and relative humidity
average 75 to 80 %). And after 10 days fermentation Sausages were packed in vacuum bags and stored at
refrigerator at 2-4ºC up to 90 days.
Microbiological Analysis
Preparation Of Samples
Microbiological Investigation For the microbial analysis, 10 g of each test were blended with 90 mL
of saline/peptone water (1 g/L peptone) and homogenized.Through mixingina stomacher 400 circulator for
Abdulrahman & Qoja (2020): Effect of microbial starter culture on fermented suasage Nov 2020 Vol. 23 Issue 18
Annals of Tropical Medicine & Public Health http://doi.org/10.36295/ASRO.2020.231817
1.5 min at 260 pm. Hence, decimal dilutions were arranged with 0.1% (wt/vol) peptone water and plated,
in duplicate, on the corresponding media (19).
Enumeration Of Total Viable Count (TVC)
Standard plate count was done as according to (20).Plate count agar has been prepared and sterilized
as per the manufacturing directions. in pre-sterilized plastic petri,one ml of each sample dilution prepared,in
the item (3.2.6.1) were inoculated, The medium was poured at 45oC of around 12-15 ml for each
plate over the sample dilution, and blended delicately until it was dispersed. The plates are incubated for
72±3 h at 30±1oC. Calculated the number and expressed as CFU/g sausage.
Lactic Acid Bacteria (LAB) Count
Lactic acid bacteria (LAB) count Man Rogosa Sharpe (MRS) Agar (OXOID, CM361) was used as
medium. Double layer cast planting. Sample Incubated at 30 oC for 72 hours at anaerobic condition (17).
Enumeration Of Total Yeast And Molds
Yeast and mold count was done as described in the international standardized NO,6611 according to
(28).DRBC agar as described by the manufacture was prepared, sterilized, and cooled to 45±1oC. The medium
was once poured nearly 12-15 ml for every plate over the sample dilution, and blended gently till it was once
dispersed. two The Plates have been incubated at 25oC for 5 days.
Statistical Analysis
Statistical analyses for all sausage samples were performed using SPSS statistical software (Version 22,
IBM), analysis factorial -Test was used to compare in between the control and fermented sausage by starter
by two factorial design, and of storage periods were evaluated by applying ANOVA according to a factorial
design with repeated measurements in time.. All figures are shown by mean values with the standard
deviation (means ± SD).
RESULTS AND DISCUSSION
Microbiological Analysisof Sausages
Total Viable Count (TVC)
Sausages total viable counting done and the number expressed as (log CFU/g). Results during the
fermentation stages of sausages are shown in Figure1. TVC numbers in the stored period is given in Figure 2.
The TVC numbers of the samples and the changes in the fermentation step versus time statistically
significant (p <0.01). Sample type x Production time also have a significant effect on TVC (p <0.01).TVC
number of the samples before fermentation was 5.44-5.88 log CFU/g for control and sausages inoculated
with starter cultures at first day. With fermentation TVC number of sausages increased and this increase was
statistically significant it was found to be important (p <0.01). On the fifth day of fermentation TVC numbers
ranged between 7.46-7.68 log CFU/g for control and sausages inoculated with starter cultures. At the study
Abdulrahman & Qoja (2020): Effect of microbial starter culture on fermented suasage Nov 2020 Vol. 23 Issue 18
Annals of Tropical Medicine & Public Health http://doi.org/10.36295/ASRO.2020.231817
on fermented sausages (21) mentioneda TVC was 5.5-6.2 log CFU/ginitially number increased to over 8.0 log
CFU/gfor fermented sausage.(17) the Italian type fermented sausages TVC 5.0 log CFU/gin the first and then
increased to reach 7.0-8.0 log CFU/gafter 14 days of the of production.
Highest bacteria number mentioned in last day of fermentation and was 8-8.37 log CFU/g for
control and sausages inoculated with starter cultures shown at Fig.1. The results agree with (22) reported that
the LAB number in the first day of production was 4.36 log CFU/gfor control sausages against 7.37 log
CFU/gfor inoculated ones also The highest level was found on the day 14 of production and then a slight
decrease was observed to reach at the end of ripening levels of 8.13 and 7.14 log CFU/grespectively for
inoculated and for control sausages.
Fig. 1: Effect of starter culture and ripening period on Sausages TVB count (log CFU/g )
TVC numbers decreased during stored period (Fig 2). Differences in TVC numbers between
samples and stored time x sample type Interaction was also statistically significant (p <0.01).The decrease in
bacterial counts of the samples was 7.9 for control sausages 8.11 sausages inoculated with starter cultures
found to be statistically significant on the 30th day.(P <0.01). On the 60th day of stored, TVC counts of
samples decreased to 6.5-6.87 log CFU/gcontrol and sausages inoculated with starter cultures. On the last day
of stored samples have the lowest TVC (5.57- 5.8log CFU/g ) control and sausages inoculated with starter
cultures respectively and at the all steps sausages inoculated with starter cultures highest log CFU/gin the
control sample shown at Fig 1 and Fig 2. Resemble with the (23) TVC numbers decreased in the sausages
stored period, and TVC at the end60th day of stored of TVC numbers was 5.0 log CFU/g.
0 Time 5 Days 10 Days
SC 5.44 7.46 8
SS 5.88 7.68 8.37
0
2
4
6
8
10
Total viable count (TVC)
(log CFU/g)
Ripening Period
Abdulrahman & Qoja (2020): Effect of microbial starter culture on fermented suasage Nov 2020 Vol. 23 Issue 18
Annals of Tropical Medicine & Public Health http://doi.org/10.36295/ASRO.2020.231817
Fig.2: Effect of starter culture and stored period on Sausages TVB count (log CFU/g)
Lactic Acid Bacteria (LAB) Of Sausages
LAB number of the samples at first day fermentation was 6.11 log CFU/gfor control 4.67 log
CFU/gand sausages inoculated with starter cultures. At the fifth days fermentation sausages numbers of
LABincreased and this increase was statistically significant it was found to be important (p <0.01). On the
fifth day of fermentation LAB numbers ranged between 7.9-7.53 log CFU/gfor control and sausages
inoculated with starter cultures. Sample type x Production time also have a significant effect on LAB (p
<0.01).Results during the fermentation stages of sausages are shown in Figure 3. LAB numbers in the stored
period is given in Figure 4.
Lactic acid bacteria in fermented meat products they ferment sugar to lactic acid (24-26)., depending
on the ripening temperature of the LAB number of sausages the second day of fermentation 3.5-4.3 log
CFU/g, and8.0 log CFU/greported at last day of fermentation the pH values the lowest level of the samples
depending on the number of LAB during this period.
(21,27) reported On the 7th day of production, LAB number that they were above 8.0 log CFU/g. (23) that the
number of sausages LAB between 5.5-7.0 log CFU/gvariancebetween them was detected. Different results
between the researches, the researchers' different starter culture and different maturing conditions may be
applied. Highest LAB number reached at in 10th day of fermentation was 8.13-8.7 log CFU/gfor control and
sausages inoculated with starter cultures.
0 Time stored 30 Days stored 60 Days stored 90 Days stored
SC 8 7.9 6.5 5.57
SS 8.37 8.11 6.87 5.8
0
1
2
3
4
5
6
7
8
9
Total viable count (TVC)
(log CFU/g)
Storage Period
Abdulrahman & Qoja (2020): Effect of microbial starter culture on fermented suasage Nov 2020 Vol. 23 Issue 18
Annals of Tropical Medicine & Public Health http://doi.org/10.36295/ASRO.2020.231817
Fig.3: Effect of starter culture and ripening period on Sausages Lactic acid bacteria count (log CFU/g )
Decrease in the number of LAB counts of samples type at stored x time Interactionwas also
statistically significant (p <0.01).The decrease in LAB counts of the samples was 8.65 log CFU/gfor control
sausages 8.1 log CFU/gsausages inoculated with starter cultures found to be statistically significant on the
30th day.(P <0.01). On the 60th day of storedLAB counts of samples decreased to 6.89-7.53 log
CFU/gcontrol and sausages inoculated with starter cultures. On the last day of stored samples have the lowest
LAB (5.1- 5.4 log CFU/g ) control and sausages inoculated with starter cultures respectively and at the all
steps sausages inoculated with starter cultures highest log CFU/gin the control sample shown at Fig 3 and 4.
The results near with (28) reported Initial counts of LAB during stored 30 days was 5,17 log CFU/gand
decreased after 60 days storedto 4,30 log CFU/g.
0 Time 5 Days 10 Days
SC 4.67 7.53 8.13
SS 6.11 7.9 8.7
0
1
2
3
4
5
6
7
8
9
10
Lactic acid acteria count
(log CFU/g)
Ripening Period
Abdulrahman & Qoja (2020): Effect of microbial starter culture on fermented suasage Nov 2020 Vol. 23 Issue 18
Annals of Tropical Medicine & Public Health http://doi.org/10.36295/ASRO.2020.231817
Fig. 4: Effect of starter culture and stored period on Sausages Lactic acid bacteria count (log CFU/g )
This slight decrease of LAB during stored is probably due to the loweringof fermentable
carbohydrates (29) and the decrease of water activity (30). LAB were the dominant microflora at the end of the
ripening for both sausages. this result confirms the good adaptation of LAB to the sausage and their faster
growth rates during production and ripening of sausages (31-38).
Sausages Yeasts and Molds Counts
Sausages yeasts and molds countsof the samples at first day fermentation was 5.32 log CFU/gfor
control 5.28 log CFU/gand sausages inoculated with starter cultures. At the fifth days fermentation sausages
numbers of yeasts and molds countsdecreased and this decrease was statistically significant it was found to
be important (p <0.01). On the fifth day of fermentation LAB numbers ranged 4.9-4.1 log CFU/gfor control
and sausages inoculated with starter cultures. These values are lower than the values of (23, 27, 39). reported the
average yeast number of sausages after drying is 6.72 log CFU/g.(23)depending on the stored temperature of
the number of yeast-mold It was between 2.5-4.5 log CFU/g.
0 Time stored 30 Days stored 60 Days stored 90 Days stored
SC 8.13 8.1 6.89 5.1
SS 8.7 8.65 7.53 5.41
0
1
2
3
4
5
6
7
8
9
10
Lactic acid acteria count
(log CFU/g)
Storage Period
Abdulrahman & Qoja (2020): Effect of microbial starter culture on fermented suasage Nov 2020 Vol. 23 Issue 18
Annals of Tropical Medicine & Public Health http://doi.org/10.36295/ASRO.2020.231817
Fig.7: Effect of starter culture and Ripening period on Sausages Moulds/yeasts count (log CFU/g)
At the 10th day this number increased to4.6log CFU/gfor control and 4.00 log CFU/ginoculated
samples. Sample type x Production time also have a significant effect on yeasts and molds (p <0.01)
Sausages yeasts and molds countsat the fermentation stages and stored period of sausages are shown in
Figure 7. In the stored period is given in Figure 8.The results agree with (22) reported that the number of
yeasts and molds increased from about 3 log CFU/gto reach about 4 log CFU/gfor both inoculated and
control sausages and was not affected by the addition of starters (p > 0.05). This evolution compare well with
most studies on fermented sausages (31,34,36,40). Different researcher Inadequateprepared sausage can become
contaminated with microorganisms due to technological processes and the use of such spices deteriorates the
quality of meat products was reported to be reduced (41).
The number of yeasts and molds counts of samples Decrease at stored from 30 to 90 dayes and
lawering count of the type at stored x and time Interactionwas also statistically significant (p <0.01).The
decrease in yeasts and molds counts of the samples was 4.11 log CFU/gfor control sausages 3.5 log
CFU/gsausages inoculated with starter cultures found to be statistically significant on the 30th day.(P <0.01).
On the 60th day of stored yeasts and molds counts of samples decreased to 3. 9-3.22 log CFU/gcontrol and
sausages inoculated with starter cultures. On the last day of stored samples have the lowest yeasts and molds
(3.78- 3 log CFU/g ) control and sausages inoculated with starter cultures respectively. The results near with
(Gök, 2006) reported Initial counts ofyeasts and molds counts during stored 30 days was 3.35 log CFU/gand
decreased after 60 days storedto 3,20 log CFU/g.
0 Time 5 Days 10 Days
SC 5.32 4.9 4.6
SS 5.28 4.1 4
0
1
2
3
4
5
6
Moulds/ yeasts counts
(log CFU/g)
Ripening Period
Abdulrahman & Qoja (2020): Effect of microbial starter culture on fermented suasage Nov 2020 Vol. 23 Issue 18
Annals of Tropical Medicine & Public Health http://doi.org/10.36295/ASRO.2020.231817
Fig. 8: Effect of starter culture and stored period on Sausages Moulds/yeasts count (log CFU/g)
CONCLUSION
The study was designed to investigatethe effect ofmicrobial starter culture on some microbial
properties of fermented sausage which obtained results show as the total viable count and lactic acid bacteria
was increased at the production period and decreased at stored period for the both type of sausages but
control counting was lower than starter inoculated sausages count and however yeast and mould was
decreased at the production period to endstored periodfor the both type of sausages but control countingwas
higher than starter inoculated sausages count.
ETHICAL CLEARANCE
The Research Ethical Committee at scientific research by ethical approval of both environmental
and health and higher education and scientific research ministries in Iraq
CONFLICT OF INTEREST
The authors declare that they have no conflict of interest.
FUNDING: Self-funding
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