Asian Food Science Journal

8(4): 1-9, 2019; Article no.AFSJ.48472

ISSN: 2581-7752

Microbial Assessment of Selected, Locally-

Fermented and Ready-to-eat Cassava
Products Sold in Lokoja, Nigeria
O. N. Akoma1*, C. M. Ononugbo1, C. C. Eze1, K. I. Chukwudozie1
and J. O. Ogwu2
1
University of Nigeria, Nsukka, Nigeria.
2
Salem University Lokoja, Kogi State, Nigeria.

Authors’ contributions

This work was carried out in collaboration among all authors. All authors read and approved the final
manuscript.

Article Information

DOI: 10.9734/AFSJ/2019/v8i429997
Editor(s):
(1) Dr. Surapong Pinitglang, Assistant Professor, Department of Food Business Management, School of Science and
Technology, University of the Thai Chamber of Commerce, Bangkok, Thailand.
Reviewers:
(1) Anna Szosland-Fałtyn, Institute of Agricultural and Food Biotechnology, Poland.
(2) New Chia Yeung, Universiti Putra Malaysia, Malaysia.
(3) Olga M. Vasek, Institute of Modeling and Innovation on Technology (IMIT), Argentina.
Complete Peer review History: http://www.sdiarticle3.com/review-history/48472

Received 20 January 2019

Original Research Article Accepted 16 April 2019
Published 24 April 2019

ABSTRACT
This study was conducted to assess the microbiological safety of locally-fermented, ready-to-eat
cassava products, namely garri and ‘fufu’, in Lokoja. A total of sixty samples comprising; twenty
white garri, twenty yellow garri and twenty fufu were subjected to microbial analysis. The samples
-4
were serially diluted to 10 and appropriate dilutions inoculated by spread plate method into
Nutrient agar, MacConkey agar and Potato Dextrose agar plates which were used for total aerobic
plate count (TAPC), coliform count (CC) and fungal count respectively. The TAPC for white garri
ranged from 0.78 to 3.83 log cfu/g, the coliform count ranged from no growth (NG) to 3.80 log
cfu/g, while the mean fungal count ranged from 1.96 to 3.39 log cfu/g. The TAPC for yellow garri
ranged from 2.04 to 3.95 log cfu/g, the coliform count ranged from NG to 3.62 log cfu/g and the
fungal count ranged from 2.08 to 3.44 log cfu/g. The TAPC of fufu was within the range of 1.07 to
3.70 log cfu/g, the coliform count ranged from NG to 3.48 log cfu/g and the fungal count ranged
from 1.94 to 2.78 log cfu/g. The bacteria isolated include Bacillus spp., Enterobacter spp.,
Pseudomonas spp., Staphylococcus aureus, Salmonella spp., Escherichia coli and Klebsiella spp.
The fungi isolated from the study samples include Aspergillus niger, Cladosporium spp., Fusarium
_____________________________________________________________________________________________________

*Corresponding author: Email: [email protected];

 Akoma et al.; AFSJ, 8(4): 1-9, 2019; Article no.AFSJ.48472

spp., Rhizopus spp., Alternaria spp., Montospora spp., and Penicillium spp. The pH of the samples
ranged from 4.02 to 4.96 in white garri, 4.02 to 4.99 in yellow garri, and 5.02 to 6.44 in fufu.
Findings showed that these widely consumed fermented (ready-to-eat) cassava products presents
(may represent) a serious risk and route for transmission of food borne pathogens to consumers
and generally huge economic disadvantage to food handlers. Improving manufacturing, packaging
and storage practices in garri production and for public health purposes are strongly encouraged.

Keywords: Microbiological safety; cassava products; microbial assessment.

1. INTRODUCTION if not properly and carefully handled during

processing and / or storage, it could serve as
Food is the basic necessity for human survival vehicle for transmission of food borne pathogens.
and attainment of food security is the priority of Moreover, there could be economic losses and
any country. However, it is important that food widespread of food borne illnesses as a result of
security should not be seen only in the contamination by these microorganisms. This
perspective of availability either quantitatively or study therefore is aimed at evaluating locally
qualitatively. Therefore, microbiological safety of fermented ready-to-eat cassava products (garri
foods should also be given important and fufu) for any microbial contamination.
consideration in order to protect the health of the Further, concerted attempt at awareness creation
people as the microbial-contaminated foods on food safety to handlers and consumers and
could serve as vehicle for the transmission of relevant contributions to the body of knowledge
food borne diseases [1,2]. Food may be available on appropriate manufacturing, packaging and
but the source from which it is produced and / or storage practices will be made.
processed may be unhygienic and even the
chemicals that may be used to preserve it may 2. MATERIALS AND METHODS
cause serious health hazard [1].
Coliforms, particularly Escherichia coli are used 2.1 Sample Collection
as indicators of post process contamination and
also the presence of E. coli in foods serves as an A total of sixty samples of locally-fermented,
indicator of faecal contamination [3]. Coliforms ready-to-eat cassava products, comprising
are group of closely related Gram negative, non- twenty white garri, twenty yellow garri and twenty
spore forming, rod-shaped aerobes and fufu were collected aseptically from the five major
facultative anaerobes that ferment lactose to markets in Lokoja Kogi state which includes;
produce acid and gas within 48 h at 35oC. They Ganaja, Adankolo, Lokongoma, Old and New
are mostly harmless and lives in soil, water and International Markets. In each of the markets,
in the gut of animals with few enteric pathogens four samples (i.e. from four vendors) of each of
including Salmonellae, Shigellae and the ready-to-eat locally fermented cassava
enteropathogenic E. coli [4]. products were collected in sterile nylon bags and
transported to Salem University Advanced
Filamentous moulds and yeasts are common Microbiology Laboratory for analyses.
spoilage organisms of food products and some
species of Penicillia and Aspergilli have been 2.2 Microbiological Analysis
reported as spoilage organisms of a variety of
foods on which they may produce a quite number Microbiological analysis was carried out
of mycotoxins [5]. according to the following conventional
Cassava ranks fourth in the list of major crops in microbiological procedures. The analysis
developing countries after rice, wheat and maize involved total aerobic plate count, fungal count
and it is used for the production of a variety of and coliform count. This was determined by the
West African foods [6]. In its natural state, it is spread plate method using standard
toxic to man as it may contain high levels of microbiological techniques.
linamarin, a cyanogenic glucoside. Hence,
processing it through fermentation enhances its 2.2.1 Enumeration of microbial isolates
detoxification, quality and safety.
Ten gram (10 g) of each sample were
Garri and fufu are the few finished products of homogenized in 90 ml sterile distilled water (10-1
fermented cassava in many parts of Nigeria, and dilution). Serial dilution of sample homogenate to

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-4
10 was carried out also in sterile distilled water 2.2.4 Isolation and Identification of bacteria
for colony count. Approximate 0.1 ml aliquot of and fungi
appropriate dilutions were spread plated on
plates of Nutrient agar and Potato Dextrose agar Pure cultures of suspected colonies were
supplemented with 0.2 μg of chloramphenicol (all obtained by repeated subculture on nutrient agar
from Bio-laboratory, Hungary). All Nutrient agar plates and potato dextrose agar plates for
and MacConkey agar plates were incubated at bacteria and fungi isolates respectively and
0
37 C for 24-48 h while all potato dextrose agar stored on slants at 4°C until characterized. The
0
plates where incubated at 25 C for 72-120 h. All isolation and identification of the bacteria were
plates were prepared in duplicate. Culture plates carried out using standard microbiological
were examined, while enumeration and techniques including; Gram stain, catalase test,
identification of colonies was carried out at the coagulase test, indole test, citrate test, oxidase
end of the incubation period. The total test [11]. All fungi isolates were identified
microbial population was expressed as log of following previously described methods [12].
colony forming unit per gram of the sample (log
cfu/g). 2.3 Determination of pH of the Locally
Fermented Ready-to-eat Cassava
Colonial morphology, microscopy and Products
biochemical tests were used to identify the
bacterial isolates [7] while the identification of The pH which is an indicator for the multiplication
characteristic fungal isolates were based on of food microorganisms was determined using
morphology and microscopy with reference to digital pH meter calibrated with standard buffer
standard atlas and keys [8]. solutions.Ten grams of each sample were
weighed and homogenized in 20ml of sterile
2.2.2 Detection of hygiene indicator distilled water in a beaker for 1 min. The solution
organisms and specific food borne decanted and the pH of the suspension
pathogens measured.

Samples of the food product were plated on 2.4 Statistical Analysis

MacConkey agar, Manitol Salt agar and
Salmonella-Shigella agar (Oxoid, England) The mean and standard deviation of the total
after pre-enrichment in Selenite F broth and viable microbial count were subjected to
0
incubated at 35 C for 24-48 h, for isolation of statistical analysis using MS Excel and SPSS
Escherichia coli, Staphylococci and Salmonellae version 20. The mean microbial load log cfug-1
respectively. and pH of the samples were presented in tables.

2.2.3 Coliform test 3. RESULTS AND DISCUSSION

Aliquot one gram samples were inoculated into The mean TAPC, fungal count, and coliform
Lactose broth in screw capped test tubes with count results in log cfu/g and respective sample
o
inverted Durham tubes and incubated at 37 C for pH are shown in Table 1. The TAPC for white
24-48 h. Tubes showing gas production and/or garri ranged from 0.78 to 3.83 log cfu/g, the
color change of dye were reported as coliform count ranged from no growth (NG) to
presumptive coliform test positive. These positive 3.80 log cfu/g, while the mean fungal count
tubes were streaked out on duplicate plates of ranged from 1.96 to 3.39 log cfu/g. The TAPC
Eosin Methylene Blue (EMB) agar for for yellow garri ranged from 2.04 to 3.95 log
confirmatory test and incubated at 37°C and cfu/g, the coliform count ranged from NG to 3.62
44°C respectively for 24 h. Growth of log cfu/g and the fungal count ranged from 2.08
characteristic colonies on EMB medium to 3.44 log cfu/g. The TAPC of fufu was within
represent confirmatory positive test which were the range of 1.07 to 3.70 log cfu/g, the coliform
Gram stained and inoculated into lactose broth count ranged from NG to 3.48 log cfu/g and the
for complete coliform test. Gas production and/or fungal count ranged from 1.94 to 2.78 log cfu/g.
color change of dye plus Gram negative non- The bacteria isolated include Bacillus spp.,
spore bearing rod represent presence of coliform Enterobacter spp., Pseudomonas spp.,
o
[9]. At 44 C absence of growth was recorded for Staphylococcus aureus, Salmonella spp.,
all the samples indicating absence of fecal Escherichia coli and Klebsiella spp. The fungi
coliforms [10]. isolated from the study samples include

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 Akoma et al.; AFSJ, 8(4): 1-9, 2019; Article no.AFSJ.48472

Aspergillus niger, Cladosporium spp., Fusarium the presence of Staphylococcus aureus, Bacillus
spp., Rhizopus spp., Alternaria spp., sp., Pseudomonas sp., E. coli and Klebsiella sp.
Montospora spp., and Penicillium spp. The pH of The observation of diverse bacteria isolates
the samples ranged from 4.02 to 4.96 in white could be attributed to the fact that these studies
garri, 4.02 to 4.99 in yellow garri, and 5.02 to was carried out at different regions and from
6.44 in fufu. different sample markets of which environmental
conditions of the study areas could affect the
The result of this study was in line with the distribution of organisms. Buyer’s attitude
earlier reported data [10,13] however, it was towards the exposed food products in the
slightly lower than another previously reported market could also contribute to the microbial
3 4
[14], with counts that ranged from 10 to 10 . load and diversity as they touch the products
Moreover, a study conducted in Ebonyi, Ogun with bare hands and taste it before they buy.
and Oyo states in Nigeria reported microbial The presence of Salmonella in this study calls
burden as high as 106 to 107 respectively [15] for concern as this organism is the common
[16], while another related work reported a cause of human food poisoning, and
4 6 -1
fungal count as high as 10 to 10 cfug [17]. salmonellosis can affect all species of domestic
The disparity in the microbial count from these animals and man. It is important to draw to our
studies could be as a result of the processing attention that the young, aged, stressed,
method, the quality of water used in the debilitated and pregnant individuals are more
production process and the length of exposure susceptible while the immunosuppressed and
during sale. A researcher equally observed back those suffering from malnutrition are at risk for
slopping used by some processors to reduce the salmonella infection [19].
length of time for fermentation to compromise
the quality of the product [16], suggesting it The presence of Bacillus and Staphylococcus
could be the lead cause of high counts observed aureus also calls for concern because some
in their study. Meanwhile, the total aerobic plate strains of these organisms are known to be
count and fungi counts of this study samples toxigenic and often implicated in food borne
were within the acceptable limit. Ready-to-eat intoxication [20,21]. Bacillus a common
foods with plate counts of 103 cfu/g are within environmental contaminant and a spore former
4 5
the acceptable limit while counts of 10 to 10 can withstand environmental stress and this may
6
cfu/g are tolerable and counts that are 10 account for its presence in the samples.
cfu/g are totally unacceptable [18]. Probable Meanwhile, Staphylococcus aureus is of human
coliform organisms were detected in most of the origin and their presence could therefore be
samples at high counts 102 to 103 cfu/g and the from the food handlers, utensils and the
presence of E. coli calls for serious concern. It environment. Moreover, garri and fufu is a
signifies poor sanitary condition and indicative of common food widely consumed by all in
faecal contamination during the production Nigeria and increasing intake of it, especially dry
process and / or storage of the fermented ready- garri as snacks or with cold water is an added
to-eat food under study. It is also indicative of practice that exposes the populace to serious
the potential presence of enteric pathogens and health risk due to the microbial status of the
therefore makes the study samples of poor product.
quality for human consumption.
Fungal isolates from the samples collected were
Bacterial isolates from the study samples were detailed in Table 4. More so, among all fungal
Bacillus sp., Enterobacter sp., Pseudomonas isolates; most authors reported the presence of
sp., Staphylococcus aureus, salmonella sp., Aspergillus sp., Fusarium sp. and Penicillium sp.
Escherichia coli and Klebsiella sp. (Table 2). in their study [10,13,17], which agrees with the
result of this study. Filamentous fungi are
Most of the isolates were glucose positive, common environmental contaminants usually
indole negative, catalase positive and a implicated in the contamination of ready-to-eat
reasonable number of both Gram positives and foods and this explains their presence in the
Gram negatives (Table 3). The isolation of study samples. More so, species of Aspergillus,
diverse microbial species from this ready-to-eat Penicillium and Fusarium are known to
fermented foods did not completely agree with produce mycotoxins [22,23,24] and their
the earlier findings [10,13,16] as each author presence in the study samples calls for serious
had dissimilarities in bacterial presence in their concern.
study samples. Most of these studies reported

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Table 1. Mean microbial count log cfu/g and pH of locally fermented cassava products in Lokoja

Sample White Garri Yellow Garri ‘Fufu’

Outlet TAPC CC FC pH TAPC CC FC pH TAPC CC FC pH
Adankolo 1 2.50±0.02 3.30±0.06 2.60±0.14 4.20 3.30±0.02 3.04±0.08 2.48±0.02 4.34 2.11±0.00 3.18±0.48 2.60±0.02 6.23
2 0.98±0.19 3.47±0.12 2.46±0.17 4.66 2.62±0.01 2.98±0.19 2.90±0.02 4.83 2.36±0.27 2.60±0.11 2.76±0.40 6.44
3 2.48±0.02 3.43±0.01 2.47±0.10 4.64 2.58±0.02 3.30±0.06 2.47±0.08 4.02 2.48±0.03 2.59±0.14 2.26±0.26 6.02
4 1.25±0.69 3.48±0.01 2.59±0.17 4.91 3.30±0.03 2.59±0.12 2.60±0.02 4.33 2.48±0.02 NG 2.66±0.26 5.93
Lokongoma 1 2.25±0.03 NG 2.60±0.03 4.36 2.50±0.06 2.60±0.03 2.85±0.02 4.54 2.30±0.06 3.14±0.07 2.00±0.06 5.64
2 2.40±0.02 3.43±0.02 2.48±0.02 4.43 2.47±0.18 NG 2.30±0.03 4.30 2.60±0.02 2.53±0.06 2.60±0.02 5.52
3 2.98±0.11 3.30±0.03 2.48±0.02 4.22 2.67±0.01 3.15±0.02 2.70±0.01 4.49 0.89±0.83 3.30±0.03 2.59±0.13 5.75
4 2.17±0.12 3.30±0.02 2.77±0.09 4.11 2.49±0.06 2.53±0.05 3.23±0.37 4.67 1.07±0.16 3.16±0.17 2.60±0.00 5.46
New 1 3.83±0.18 NG 3.39±0.40 4.40 2.11±0.05 3.72±0.34 2.59±0.16 4.52 2.50±0.06 3.43±0.02 2.48±0.03 5.62
2 0.78±0.67 3.80±0.27 2.60±0.03 4.43 2.20±0.06 3.00±0.06 2.77±0.08 4.23 3.00±0.09 3.48±0.03 2.48±0.02 5.41
3 2.58±0.03 3.30±0.08 2.47±0.15 4.59 3.95±0.07 2.90±0.12 2.95±0.03 4.12 2.64±0.06 NG 2.78±0.05 5.03
4 2.00±0.06 3.45±0.21 2.60±0.01 4.17 2.08±0.05 2.30±0.06 2.60±0.02 4.04 3.48±0.02 3.43±0.01 2.48±0.01 5.14
Old 1 3.00±0.06 2.89±0.04 2.60±0.02 4.08 2.04±0.06 2.30±0.02 2.70±0.06 4.50 3.00±0.03 3.30±0.09 2.60±0.03 6.35
2 2.08±0.05 3.85±0.00 2.47±0.10 4.87 2.17±0.08 2.46±0.19 2.95±0.01 4.12 2.08±0.05 3.30±0.05 2.47±0.10 6.23
3 2.11±0.01 2.48±0.03 2.59±0.17 4.96 2.17±0.24 NG 2.70±0.01 4.73 1.90±0.43 2.47±0.10 1.94±0.34 6.10
4 3.76±0.16 NG 2.60±0.03 4.15 3.69±0.12 2.00±0.06 2.00±0.12 4.94 3.24±0.34 NG 2.75±0.21 6.06
Ganaja 1 2.07±0.10 2.30±0.06 2.48±0.02 4.96 2.22±0.25 3.62±0.04 2.60±0.05 4.56 2.02±0.17 2.29±0.16 1.94±0.34 5.64
2 2.28±0.07 2.24±0.34 1.96±0.26 4.02 2.10±0.14 3.49±0.06 3.44±0.27 4.15 2.15±0.21 3.22±0.38 2.45±0.21 5.43
3 2.17±0.12 NG 2.75±0.21 4.14 2.05±0.21 1.95±0.49 2.66±0.26 4.77 2.20±0.12 2.90±0.02 1.98±0.19 5.02
4 2.30±0.03 NG 2.00±0.12 4.25 3.68±0.19 NG 2.08±0.05 4.99 3.70±0.02 2.24±0.34 2.75±0.21 5.13
Key: TAPC = Total Aerobic Plate Count, CC = Coliform Count, FC = Fungal Count, NG = No Growth

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Table 2. Distribution of bacteria in the food samples across the sampled markets

Market Sample Salm Kleb Staph Bacillus E. coli Strept Pseudo Entero
Adankolo White garri + + + - - - - -
Yellow garri - - + + - - - -
Fufu - + + - - - + -
Lokongoma White garri + - - - + - - -
Yellow garri + - + + - - - -
Fufu - - + + - - - -
New White garri - - + - + + - -
Yellow garri + - + - + - - -
Fufu - + + - - + - -
Old White garri + - + + + - - -
Yellow garri - - + - + - - +
Fufu + - + - - + - -
Ganaja White garri - - + - + - - -
Yellow garri - - + - + - - -
Fufu - - + - - - + -
Key: Salm = Salmonella sp., Kleb = Klebsiella sp., Staph = Staphylococcus aureus, Pseudo = Pseudomonas sp., Entero = Enterococcus sp., + = present, - = absent

Table 3. Biochemical characteristics of bacterial isolates

Bacteria found Gram stain H2O2 Glucose Citrate Indole Endospore Oxidase Coagulase Catalase
Salmonella sp. - ND + - - - - - +
Klebsiella sp. - ND + + - - - - +
S. aureus + ND + + - - - + +
Bacillus sp. + ND + + - + + ND +
E. coli - ND + - + - - - +
Streptococcus sp. + ND + ND ND - ND - -
Pseudomonas sp. - - - + - - + - +
Enterococcus sp. + - + - - - - - -
Key: ND= Not determined

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Table 4. Distribution of fungi in the food samples across the sampled markets

Market Sample Asper Peni Mold Mucor Rhi Fus Clado Alter Monto
Adankolo White garri + - - - - - - - -
Yellow garri + + + - - - - - -
Fufu - + - - - - - - -
Lokongoma White garri - + - + - - - - -
Yellow garri - - - - + - - - -
Fufu - - - - - + - - -
New White garri - + + + - - - - -
Yellow garri - - + + - + - - -
Fufu - + - - + - - - -
Old White garri - - - + + - - - -
Yellow garri - - - - - - + + -
Fufu - - - - - + - - -
Ganaja White garri + - - - - + - - -
Yellow garri - - - + - - - - +
Fufu - - - + - - - - -
Key: Asper = Aspergillus sp., Peni = Penicillium sp., Rhi = Rhizopus sp., Fus = Fusarium sp., Clado = Cladosporium sp., = Alter = Alternaria sp., Monto = Montospora sp.,
+ = present, - = absent

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The pH value of white garri and yellow garri among faecal coliforms and Escherichia
ranged from 4.02 to 4.96 and 4.02 to 4.99 coli in various foods. European Food
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