Authors : Ekwi Damian N; Bertrand Tatsinkou Fossi; Léopold Tatsadjieu Ngoune; Ebot Sahidu Tabe
Volume/Issue : Volume 6 - 2021, Issue 9 - September
Google Scholar : http://bitly.ws/gu88
Scribd : https://bit.ly/3Buf2EQ
Traditionally fermented foods such as, ‘Corn
Pap’ (fermented corn paste), “Kum-kum” (fermented
cassava-derived powder), ‘Water fufu’ (Fermented
cassava-derived paste), and Soya beans yogurt
(processed from fermented soya beans), have an old
history in Cameroon and are staple foods largely
consumed on daily basis by majority of both young and
elderly people in the main cosmopolitan cities such as
Yaounde, Douala, Bamenda and Buea, where majority
of the ethnic groups in Cameroon reside. Lactic acid
bacteria (LAB) have been used in the production of
fermented foods and bacteriocins produced by these
bacteria are harmless and have the ability to inhibit the
growth of pathogenic bacteria. The aim of this study was
to screen and identify genotypically bacteriocinproducing lactic acid bacteria from locally fermented
foods in Cameroon against two food born Salmonella
enterica pathogens and to assess the phylogenetic
relationship among the isolates.
A total of 112 fermented food samples were
obtained randomly from different local foods markets in
the four main cities in Cameroon. All the samples were
cultured in the De man, Rogosa, and Sharpe (MRS)
medium which supports the growth of lactic acid
bacteria. A total of 70 isolates were found to be Grampositive and catalase-negative and were further
identified by using polymerase chain reaction (PCR)
targeting the 16S rRNA gene. Genomic DNA of bacteria
isolates were extracted using EZ-10 spin column genomic
DNA Extraction kit and amplification of 16SrRNA gene
of bacterial isolates was done using the bacteria
universal MP096 forward and reverse primers and
sequencing was carried out at Genewiz, NJ, USA.
Selection for DNA sequencing was based on their
bacteriocin-producing potentials which produced
inhibition zones on culture plate. Sequence analysis was
carried out using BLAST program and comparison with
the resources at the NCBI database. Phylogenetic tree
was constructed to determine the closest bacterial species
and strains by Neighbour Joining method using MEGA
X software, version 10.0.5.
Results showed that 70 PCR-amplified products
were obtained which revealed clear single bands of
presumed LAB isolates on 1.5% (w/v) agarose gel with
average molecular weights in the range of 800bp-1300bp.
The sequencing results identified 23 lactic acid bacteria
strains (34.9%) which had similarity over 90% with
those deposited in the NCBI GenBank. The isolates were
identified as; L. plantarum (9 strains), Uncultured L. spp
(4 strains), L. paraplantarum (3 strains), L. pentosus (2
strains), L. salvarius (2 strains), Pediococcus acidilactici
(2 strains) and 1 strain belong to Pediococcus cellicola.
There was a significant difference (P<0.05) in the
number of isolates obtained from Corn pap compared to
the other fermented food sources. The highest number of
isolates were obtained from Corn pap sample (9),
followed by 5 each from Kumkum and Soyabeans, and 4
from Water fufu samples. Results also showed genetic
relationship of isolates from Yaounde, Douala and
Bamenda to have ancestry from isolate obtained from
Buea.
Therefore, this study has shown the presence of
different LAB bacterial strains in the fermented foods
and constitute potential sources of bacteriocinogenic
LAB against Salmonella Typhimurium and S.
Enteritidis. The distribution of the strains from
Bamenda, Douala and Yaounde, were extremely likely to
originate from the strain, Lactiplantibacillus plantarum
strain 07B144, obtained from Buea.
Keywords : Bacteriocinogenic; Isolation; Lactic Acid Bacteria; Molecular Identification; Phylogenetic Analysis; Salmonella Enterica; Traditionally Fermented Foods.