Chapter 6

Microbiological composition of
Tayohounta , a fermented baobab
flavour food of Benin

This chapter has been submitted as

Microbiological composition of Tayohounta, a fermented baobab flavour food of Benin
Chadare F.J., Jonkman J., Wolkers-Rooijackers J., Nout M.J.R., Hounhouigan J.D. and
Zwietering M. (submitted)

3
Microbiological composition of
Tayohounta

4
 Microbiological composition of
Tayohounta

Abstract
The importance of forest foods in general and in particular baobab foods in most African
countries is well known. The present work provides data on the microbiology of Tayohounta,
a product from natural fermentation of baobab seeds kernels from Benin. Samples were
produced and collected from 3 different small scale producers from Benin at the end of the
fermentation process. Aerobic mesophilic microorganisms, lactic acid bacteria (LAB),
Enterobacteriaceae, bacterial spores, micrococci, yeasts and moulds were enumerated and
identified using phenotypic and genotypic methods. Strains were further investigated using
Denaturing Gradient Gel Electrophoresis (DGGE) and cloning techniques. Isolated
microorganisms were tested for their effect on pH, protein degradation and smell as an index
of their functionality in baobab seed kernels fermentation.
Whereas B. subtilis caused a degradation of protein and developed a lower aroma intensity,
Enterobacteria and lactic acid bacteria (LAB) had little impact. On the other hand, a few
selected moulds and yeasts caused pH increase, protein degradation and off-flavour. Total
viable counts (TVC) were around 9 log cfu/g consisting mainly of Bacillus spp., whereas
LAB (8 log cfu/g) and yeasts and moulds represented a minor part of the total flora in all
samples. Identification of isolated strains showed that samples produced by different
producers may have different microbial composition. However, it could be shown that there
is a common presence of Bacillus subtilis, Bacillus licheniformis, Bacillus
thermoamylovorans, Lactobacillus fermentum and Streptococcus spp., whereas other
Bacillus spp., Enterococcus, Lactobacillus and Pediococcus spp. were found in products
from two of the three producers. Many other identified species (i.e., Klebsiella pneumoniae,
Lactobacillus agilis, Staphylococcus aureus) were found in only one of the 3 Tayohounta
samples.

5
 Microbiological composition of
Tayohounta

Keywords: Adansonia digitata, natural fermentation, Bacillus spp., DGGE, genotyping,

functionality

6
 Microbiological composition of
Tayohounta

6.1 Introduction

In many African countries, the baobab (Adansonia digitata L.) is a widely used tree for many
purposes especially as food and therapeutic product. All parts of the tree (roots, bark, leaves,
fruits and seeds) are used as ingredients in the preparation of traditional foods (Nordeide et
al., 1996) and some have a therapeutic effect (Codjia et al., 2001). The most common
baobabderived foods are juices from the pulp, powdered leaves used as an ingredient for
sauces, and flavouring agents made from the seeds (Sidibe and Williams, 2002). Up till now,
data are available on the ethnobotanic, ecological and genetic aspects of the baobab tree, as
well as on the chemical composition of baobab parts (Nour et al., 1980; Ajayi et al., 2003;
Osman, 2004; Assogbadjo et al., 2005; Assogbadjo et al., 2006). Recent concern about the
lack of documentation about traditional and indigenous food cultures, which is important to
retain traditional food knowledge and sustainable food systems, has lead towards a focus on
African food cultures (Wahlqvist, 2007). Research on traditional African foods can provide
possibilities for valorization through improvement of traditional techniques and products
leading to production of added value products for a larger market and a better price. This has
yielded valuable information (physico-chemical composition and microbiology of the foods)
about some West-African traditional foods in general and fermented ones in particular, e.g.,
fermented cereal products such as mawe, gowe, ogi (Hounhouigan et al., 1993; Agati et al.,
1998; Michodjehoun-Mestres et al., 2005), and fermented seed products such as afitin, sonru,
iru, kpaye, ogiri, ugba, dawadawa and soumbala (Omafuvbe et al., 1999; Sanni et al., 2000;
Dakwa et al., 2005; Azokpota et al., 2006). Some microorganisms present in the above cited
fermented products have shown to be functional, i.e. they contribute to desirable outcomes of
fermentation. Most of the fermented seed products are used as flavouring agents; therefore,
the flavour is desirable and hence functional in those products. Volatile compounds resulting
from the metabolic activities of Bacillus spp. contribute significantly to flavour during the
fermentation of African locust beans to produce soumbala (Ouoba et al., 2005) or soya beans
to produce natto or thai thua-nao (Leejeerajumnean et al., 2001). As such, many volatile
compounds were found in the Beninese afitin, iru and sonru (Azokpota et al., 2008). Some
Bacillus strains also showed to be functional in kinema and soumbala for protein degradation,
pH increase and development of desirable stickiness (Sarkar et al., 2002). Such works have,
however, never been performed on any of the baobab products.
In relation to baobab, a study was carried out on the possibility to incorporate baobab pulp in
tempe, a fungal fermented soybean food (Afolabi and Popoola, 2005). In Benin, thirty five
baobab food products have been recorded among which several fermented products which
were not yet characterized before (Chadare et al., 2008).
Tayohounta is one of those indigenous fermented baobab foods from Benin, and its
production process was described recently (Chadare et al., 2010). It is a product belonging to
the category of the alkaline fermented foods (Steinkraus, 1995) and it is used as a source of
flavour when making soups. There is some diversity of Tayohounta from different producers,
possibly because of the uncontrolled natural fermentation taking place, in which high
numbers of Bacillus spp. were observed. This would give it resemblance to other regional
flavour products such as afitin, iru and soumbala. In order to better understand and
characterize the common microflora and arrive at a basis for the future development of
fermentation starter cultures for Tayohounta, the present analysis of the product of three
individual small-scale producers was carried out.

7
 Microbiological composition of
Tayohounta

6.2 Materials and methods

6.2.1 Sampling

Tayohounta production process was followed at 3 representative small-scale production sites

in Boukoumbe, North-Benin. The production process was as described by Chadare et al.
(2010). Briefly, baobab seed kernels were roasted (5-10 min), cooked in water (about 30
min), transferred in a plastic container, covered with leaves (usually leaves of Annona
senegalensis) and allowed to ferment for 2 days at 24-31°C (night-day ambient temperature).
Cover leaves are in contact with the product. Processor 3 cooked for a longer time and used
different leaves to cover than processors 1 and 2. Samples were collected just at the end of
the fermentation period and stored in refrigerated conditions (± 4°C) for one week prior to
analysis. Visual comparison of the samples prior to analysis showed that samples from
processor 1 and 2 still had the structure of intact seed kernels with a dry or dull surface while
the sample from processor 3 showed more decomposition of the kernel structure and had a
moist appearance. Also the flavour of sample from processor 3 seemed much more pungent
than the ones of the two other samples.

6.2.2 Enumeration of the groups of microorganisms

Representative sub-samples of 10 g were weighed aseptically and were made into 10 -1

dilutions and consecutive 10-fold dilutions, using peptone physiological saline (PPS) solution
(8.5 g NaCl and 1 g neutralized bacteriological peptone (Oxoid, LP0034) in 1 L
demineralised water) in sterile filter Stomacher bags. The samples were homogenized for 60
seconds with a Stomacher (Seward Laboratory Blender Stomacher 400) at normal speed.
Aerobic mesophilic bacteria were enumerated on plate count agar (PCA, Oxoid CM0325)
(30°C, 2 days) and reported as Total Viable Count (TVC).
Lactic Acid Bacteria (LAB) were enumerated on de Man, Rogosa and Sharpe Agar (MRSA,
1.5% Technological Agar (Oxoid, LP0013) added to de Man, Rogosa and Sharpe broth
(Merck, VM986641, mixed and sterilized) supplemented with natamycin (2 g Delvocid (50%
natamycin, DSM) added to 1 L MRSA). After solidification the plates were stored in an
airtight jar under microaerophilic conditions (80% N 2, 10% CO2 and 10% H2 gas mixture
leaving a final concentration of O2 of 6%) and incubated at 30°C for two days. Presumptive
LAB were confirmed by oxidase and catalase tests, and confirmed counts were reported as
Lactic Acid Bacteria (LAB).
Enterobacteriaceae were enumerated on Violet Red Bile Glucose (VRBG) agar (Oxoid,
CM0458), in microaerobic conditions (30°C, 24 h).
Bacterial spores were enumerated on PCA (30°C, 2 days), after heating the 10 -1 dilution at
80°C for 10 min to kill vegetative cells. Micrococci were counted on Mannitol Salt Agar
(MSA, Oxoid CM0085) (30°C, 2 days). Yeasts and moulds were enumerated on
Oxytetracycline Glucose Yeast Extract Agar (OGYEA, Oxoid CM0545 supplemented with
oxytetracycline (25°C, 3 days). Yeast and mould colonies were counted separately (Nout,
1991).

6.2.3 Isolation, purification and confirmation

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 Microbiological composition of
Tayohounta

Distinctive colonies were isolated (5 per group of microorganisms and per sample), from the
counted plates to confirm the obtained counts by microscopic observation and biochemical
analysis, identification and experimental fermentations. Colonies were purified by streaking
on adequate medium, incubated at the required temperature, and further stored on slants kept
at 4-7°C.
Wet-mount preparations of the isolated strains were made and observed under a microscope
(Olympus, BX40). A preliminary grouping was based on morpholo ical traits (cell shape and
size).

DNA extraction and amplification from isolated bacteria. From the confirmed isolates, DNA
was extracted using the ‘Isolation Genomic DNA for Gram positive and Gram negative
bacteria’ kit A1125 from Promega, Southampton, UK, according to the manufacturer’s
instructions. The DNA was stored at 4°C. The DNA isolated for bacterial isolates was used to
amplify the 16S rDNA gene by polymerase chain reaction (PCR), using forward primer
5’AGA GTT TGA TCC TGG CTC AG-3’ and reverse primer 5’- AAG GAG GTG ATC CAG
CCG CA-3’ (Oomes et al., 2007). The PCR was done using a GeneAmp PCR system 9700
(Applied Biosystems) with PCR conditions as follows: initial denaturation of double-stranded
DNA for 5 min at 94°C; 35 cycles each consisting of 30 s at 94°C, 20 s at 56°C, and 1 min at
72°C; and extension of incomplete products for 7 min at 72°C followed by cooling at 4°C.
The PCR products were sent to GATC Biotech (http://www.gatc-biotech.com) in Germany
for purification and sequencing.

DNA extraction and amplification from isolated yeasts and moulds. Cultures were grown in
MEB at 30°C for 1 day for yeasts and for moulds at 25°C for 2 days and then centrifuged at
6000 rpm for 10 minutes at 4°C, followed by washing the pellets with PBS buffer and
centrifuged again as mentioned above. After the second centrifugation the pellets were then
resuspended in 0.5 mL PBS buffer and transferred into 2 mL eppendorf tubes containing 0.5 g
zirconia/silica beads (diameter 0.1 mm; Biospec products, Inc). Furthermore 0.5 mL CTAB
and 0.5 mL phenol-chloroform-isoamyl alcohol (25:24:1) were added and tubes were beaten
at maximum speed twice per 45 s using Mini beadbeater-8, Biospec, Bartlesville, USA.

Amplification and sequencing of fungal DNA. The DNA isolated from fungal isolated
microorganisms was used to amplify the 26S rDNA using forward primer V9G (5’-
TTACGTCCCTGCCCTTTGTA-3’) and reverse primer LS266 (5’-
GCATTCCCAAACAACTCGACTC-3’) (van den Ende and de Hoog, 1999). PCR was
performed with a total reaction volume of 50 μl containing 26.6 μl ddH2O, 5 μl PCR buffer, 3
μl MgCl2 (25 mM), 10 μl dNTP (2 mM), 2 μl of each primer (10 μM), 1 μl DNA template,
and 0.4 μl Taq DNA polymerase (5 U/μl) (Fermentas, USA). PCR was done using GeneAmp
PCR system 9700 (Applied Biosystems) with PCR conditions as follows: initial denaturation
for 5 min at 95°C followed by 35 cycles comprising denaturation at 95°C for 60 s, annealing
was at 52°C for 45 s, extension at 72°C for 60 s, and final 7 min extension at 72°C followed
by cooling to 4°C. The PCR products were sent to GATC Biotech
(http://www.gatcbiotech.com) in Germany for purification and sequencing.

DNA extraction directly from Tayohounta samples. Four grams of sample were suspended and
DNA was extracted according to Wang et al. (2008). From the DNA extracted directly from
Tayohounta the V6-V8 region of the 16S rDNA gene was amplified using the universal

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 Microbiological composition of
Tayohounta

forward primer EUB-968-GC-for (5'-CGC CCG GGG CGC GCC CCG GGC GGG GCG
GGG GCA GGG GAA CGC GAA GAA CCT TAC-3') and reverse primer EUBL1401-rev
(5'-CGG TGT GTA CAA GAC CC-3 (Engelen, Heuer et al., 1995).

Cloning of PCR products. Identification of microbial strains was obtained by cloning PCR
fragments in E. coli according to Röling et al. (2001) and sequencing 96 out of 200
transformants.

DNA sequence analysis. Obtained sequences were viewed for errors using Chromas Lite
V2.01 software (http://www.technelysium.com.au/) and if necessary corrected manually with
the IUB/IUPAC nucleic acid code. Afterwards the forward and reverse sequence were
assembled to form a contig using the program Seqman II (DNAStar Inc. USA, version 5.08)
and exported to FASTA format. These files were used to obtain strain identities by using the
nucleotide BLAST program from the National Center for Biotechnology Information
(http://www.ncbi.nlm.nih.gov/). The most probable match with a 16s rDNA sequence was
selected based on the percentage of identification (%ID).

Denaturing Gradient Gel Electrophoresis: PCR products were analyzed by DGGE as

described by Wang et al. (2008). 4-10 μg DNA was applied and a range of 30-60% denaturant
was used. The gel was run for 10 min at 200 Volts and 60°C, followed by 16 h at 85 Volts and
60°C. Then, the voltage was switched down to 10 Volts and run until the gel was removed
from the buffer. Silver staining was done according to Sanguinetti et al. (1994). The dried gel
was scanned on a GS 800 calibrated Densitometer (BioRad). The program Bionumerics
(Applied Maths) was used to normalise the gel and analyse band positions. Band positions
from the pure cultures were compared to the pattern given by mixtures of the pure cultures
and of the original samples.

6.2.4 Laboratory-scale fermentation of Baobab seeds

A selection of microorganisms isolated from Tayohounta was used as fermentation starters

and their effect was measured. A quantity of 1000 g dry seeds was soaked in 2 litres of tap
water overnight at 25°C. After pouring off the water, 50 g aliquots of the soaked kernels were
transferred into 500 mL glass jars and sterilized by autoclaving at 121°C for 30 min. The
seeds were inoculated with pure cultures, pre-grown for 48 h at 30°C in tubes containing 10
mL of nutrient broth (Bacillus), MRS broth (LAB) and malt extract broth (MEB; Oxoid
CM57) (yeasts and moulds). One mL of a 10 -1 dilution in PPS was used to inoculate each jar
of seeds, and these were incubated at 30°C for 0 h (unfermented control) and 48 h.

Functional properties
Microbial growth. Each sample of fermented kernels was analyzed as described in section
6.2.2.

Terminal amino Nitrogen and pH. Ten grams sample was mixed with 90 g water and made
into a suspension with a blender. This suspension (A) was used to measure pH and to perform
the formol titration (Han et al., 1999) for terminal amino groups, which were expressed as
mM/g sample.

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 Microbiological composition of
Tayohounta

Volatile flavour: Of each fermented or non-fermented sample the flavour was examined by
sniffing unfermented as well as fermented seeds. This was done by 3 untrained panelists.

6.3 Results and discussion

6.3.1 Enumeration of groups of microorganisms

Table 6.1 shows that the TVC in all samples was quite high, around 9 log cfu/g. High
numbers were expected because there was no process step performed on the samples which
would decrease the number of microorganisms after fermentation. The groups of LAB and
bacterial spores, with counts around 8 log cfu/g, represented a large part of the TVC, but
considering the high number of spores it is probable that the remaining TVC consisted mainly
of vegetative cells of Bacillus spp. Yeasts and moulds, represented only a minor part of the
total flora (Table 6.1).

Table 6.1 Microbiota composition of Tayohounta made by three individual processors

(Log cfu/g) Tayohounta 1 Tayohounta 2 Tayohounta 3

Total Viable aerobic count 9.0 ± 0.1 a 8.8 ± 0.1 a 8.7 ± 0.1 a
Lactic acid bacteria 7.7 ± 0.0 c 8.5 ± 0.0 a 8.2 ± 0.0 b
Enterobacteriaceae 6.0 ± 0.4 a 7.0 ± 0.0 a <1
Bacterial spores 8.0 ± 0.0 b 8.5 ± 0.1 a 8.1 ± 0.0 b
Micrococci 7.2 ± 0.1 a 5.1 ± 0.1 b 5.3 ± 0.2 b
Yeasts 7.2 ± 0.0 a 3.1 ± 0.1 c 5.9 ± 0.0 b
Moulds 2.9 ± 0.1 a 2.4 ± 0.1 b <1

For each group of microorganisms, samples with different letters are significantly different (p < 0.05)

Substrate depletion by the rapid growth and high numbers of bacteria, as well as formation of
metabolites such as ammonia, might retard the growth of yeast and moulds present. Because
no starter is added after the seeds are boiled, likely origins of the microorganisms observed
include kitchen utensils, cover leaves and other environmental sources (soil, water and air). It
can also be possible that some spores were still viable after the long cooking procedure which
correspond to moist heat treatment (Melly et al., 2002; Coleman and Setlow, 2009) or that the
spores had formed clumps which increase their heat resistance (Furukawa et al., 2005).

6.3.2 Identification of isolated strains

Table 6.2 shows the identification of bacteria in Tayohounta made by 3 processors, based on
sequencing 16S rDNA of isolated pure cultures, and on cloning DNA products obtained by
PCR amplification from DNA extracted directly from Tayohounta samples.
The data presented in table 6.2 support the observation that products 1 and 2 show more
similarities and that product 3 is different. Cloning techniques were therefore performed only
on T1 and T3 (Figure 6.1); however, results from all the used techniques and thus on T1, T2
and T3 are often integrated in the interpretation. The identification of the microorganisms
based on clone library of PCR product, DGGE and isolated pure culture (Table 6.1 and

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 Microbiological composition of
Tayohounta

Figure 6.1) shows that Bacillus subtilis, Bacillus licheniformis, Bacillus thermoamylovorans,
Lactobacillus fermentum and Streptococcus spp., were present in T1, T2 and T3 while some
strains were found in products from two of the three producers; they are: Enterococcus
casseliflavus (T1 and T2), Pediococcus pentosaceus (T1 and T2) and Bacillus cereus (T1 and
T3). Many other identified species (i.e., Bacillus thurengiensis, Brevibacillus borstelensis,
Klebsiella pneumoniae, Lactobacillus agilis, Staphylococcus aureus) were found in only one
of the 3 Tayohounta samples. Strains of Bacillus subtilis represent 40% of all strains in
product 3 and 27% of the ones in product 1. These two samples are very different while
considering the other strains. Product 1 contains Enterococcus durans (22%) and
Streptococcus sp. (20%) which were less abundant in product 3. On the other hand, Bacillus
thermoamylovorans is more represented in product 3 (33%) than in product 1 (4%).
Pediococcus pentosaceus and Bacillus pumilus were found in products 1 and 2, confirming
their stronger resemblance. All other species identified were only present in one of the three
samples. Research done elsewhere on similar products also identified B. subtilis as
dominating organism within the Bacillus group, sometimes even responsible for 50% of the
total Bacillus spp. count, often accompanied by B. licheniformis, B. pumilus and B. cereus
(Omafuvbe et al., 1999; Dakwa et al., 2005; Azokpota et al., 2006). Heat resistance of B.
subtilis spores has been investigated by Fox and Eder (1969) and Warth (1978), who found
that the decimal reduction time around 100°C (D 100) was in the order of magnitude of 10 min.
More recently, Oomes et al. (2007) reported heat resistance of B. subtilis in nutrient broth
with added mineral mix to range widely from D 111 = 0.5 min to D121 = 7.9 min. Considering
that the boiling step during the process can take up to half an hour it seems quite possible that
certain Bacillus spores can survive the boiling procedure when present in high numbers.
Because there are differences in appearance and smell between the samples it can be stated
that next to B. subtilis the other organisms play an important role during fermentation of the
seed kernels as well. Again a different flora of the environment and leaves or on the skin of
the producers and used utensils is suggested as probable causes for variability of microbiota.

6.3.3 Functionality

Results of functionality test of the microorganisms during the fermentation of baobab seed
kernels into Tayohounta are presented in table 6.3. All tested strains showed good growth
within 48 h of incubation.
Only a few selected isolates for each group of microorganisms were tested for their effect on
quality attributes of fermented kernels. Bacillus subtilis isolates showed no effect on pH, but
were able to degrade protein and caused a typical pungent smell. Enterobacter and Klebsiella
spp. had no effect on pH, protein or smell. Pediococcus sp. caused a slight decrease of pH,
slight degradation of protein, and minor change of smell. The strongest effects were caused
by Aspergillus sp. and two unidentified yeast isolates, which caused a rather unpleasant
smell. In the fermentation of soybeans by Bacillus strains for preparation of Soumbala and
Kinema, a correlation was observed between increasing pH and level of free amino-N (Sarkar
et al., 2002). In the present study, though Bacillus strains were able to degrade protein,
increase of free amino-N was not correlated with increasing pH in the fermentation of baobab
kernels; yeasts and moulds were capable of producing volatile flavour. The reproducibility of
the used technique for smell detection might be relatively low and was subjective; it is

12
 Microbiological composition of
Tayohounta

important to investigate the formation of volatiles more in detail before accepting or rejecting
potential fermentation starters.

13
 C C C I I
Processor 2 Processor 3 C1
2 1 1

on cloning DNA products obtained by PCR amplification from DNA extracted directly from
I I I
samples I; C 40
I; C2 C 33 I; C 1
C4 C1
Tayohounta

I; C 27 I
I; C5 I C5I I; C 1 I
C4 C4 C 22 C 1C 1 C 20
Processor 1

I : based on isolated pure culture; C : based

I I I
Bacillus thermoamylovorans
Enterococcus casseliflavus Jeotgalicoccus halotolerans
Brevibacillus borstelensis Lactobacillus
Pediococcus pentosaceus
fermentum
Klebsiella pneumoniaefaecium
Enterococcus Staphylococcus aureus
Bacillus thuringiensis
Bacillus licheniformis Enterococcus
Enterococcus
Enterobacter cloacae duransitalicus
Lactobacillus agilis
Lactobacillus equi confusa
Weissella
Bacillus pumilus
Bacillus subtilis of total clones identified (96 clones tota
Bacillus cereus
Tayohounta Bacillus sp. Streptococcus
sp.

Identification of bacteria in
Name

Sporeforming bacteria
Lactic acid bacteria
Enterobacteriaceae

Micrococci
Group
Table 6.2

Figure 6.1 Clone library of PCR product from direct extraction of sample made by 2 processors. T1,
T3 = sample made by processor 1 and 3, respectively
 Microbiological composition of
Tayohounta

Legend to marker bands:

1: Pediococcus pentosaceus
2: Streptococcus sp.
3: Bacillus thuringiensis
4: Streptococcus equinus
5: Lactobacillus agilis / Lactobacillus equi
6: Bacillus subtilis
7: Bacillus pumilus
8: Bacillus subtilis
9: Bacillus subtilis/Bacillus licheniformis
10: Bacillus licheniformis
11: Brevibacillus borstelensis
12: Enterococcus casseliflavu s / Enterococcus durans
13: Bacillus subtilis
14: Enterococcus durans
15: Bacillus subtilis
16: Bacillus subtilis / Bacillus licheniformis
17: Bacillus subtilis
18: Lactobacillus fermentum
19: Bacillus licheniformis
20: Bacillus thermoamylovorans

15
 Microbiological composition of
Tayohounta
Chapter 6

6.4 Conclusion

There is evidence of considerable microbial and quality diversity of Tayohounta made by

different traditional processors using different ingredients. Similar to most fermented seed
products (afitin, iru, sonru, dawadawa, etc.), Bacillus subtilis and other Bacillus species
are present in Tayohounta whatever the process/producer. It also became clear that the
culture-dependent analysis (isolation of pure cultures) and culture-independent approaches
(direct DNA extraction from Tayohounta) resulted in overlapping results. However,
cloning of PCR products yielded species that had not been isolated, while some isolated
species were not discovered by cloning. This confirms earlier findings (Martin Platero et
al., 2008) that a polyphasic approach is the most suitable one for studying the complex
microbiota of naturally fermented foods.

16
 Microbiological composition of
Tayohounta
128

17
 Microbiological
penetrativecomposition
sour of Tayohounta
Cooked pasta, but

Cooked
Cooked
Cooked
Cooked Cooked
Cooked
Cooked
Cooked
Cooked
Aba
less strong than ym
less
9 strong than
t = 0 h t = 48 h smoky nuts
wet bread
wet bread wet bread
t = 0 h t = 48 h t = 0 h t = 48 h t = 0 h t = 48 h Increase
Smell older beans cream
0.03 0.06
0.02 2.4
0.09
0.02 3.8
0.02
0.03 1.0
0.03
0.02 1.0
0.04
0.03 1.8
0.04
0.02 1.4
0.05
0.03 2.2
0.04
0.02 2.0
0.05 2.1
pasta acidic
during fermentation of cooked baobab seeds

pasta pasta pasta pasta pasta pasta pasta pasta

Terminal Amino Groups

factor

mmol
)
-1 g
(
pH

5.8

Tayohounta,
Growth and impact of selected pure isolates from
5.9 5.9 5.9 5.9 5.7 5.8 6.1 6.6 6.4
6.3* 8.8
6.3 5.9
9.2
6.35.9
8.2 5.9 6.3 7.5
6.35.8
7.6
4.35.8
7.3
5.35.8
7.3
5.35.9
7.1 5.9
ND

Code Log cfu g d from inoculation rate

6.3

-1

*Approximate values calculate

Unidentified yeast Dym

bs bs LAB LAB ym ym
Aba Bba 16 17 3 4 9 10

Unidentified yeast
Bacillus
Bacillus subtilis Pediococcus
subtilis Pediococcus
sp. sp.
Aspergillus sp.
Enterobacter
pneumoniae
Table 6.3 Klebsiella
Species cloacae

18
Microbiological composition of Tayohounta

19