UJMR, Volume 4 Number 2, December, 2019, 43 - 48 ISSN: 2616 – 0668

https://doi.org/10.47430/ujmr.1942.008

Received: 04/05/019 Accepted: 23/06/2019

Identification and Anti-bacterial Testing of Staphylococcus aureus Isolated from Jollof
Rice sold at selected Cafeterias in Federal University Dutse Campus, Jigawa State.
1
Raji, M., 1Amoo, F. K., 2*Adeleye, A. O., 2Amoo, A.O., 2Bate, G. B. and 1Mohammed, S.
1
Department of Microbiology and Biotechnology. Federal University Dutse, Jigawa State.
2
Department of Environmental Sciences, Federal University Dutse, Jigawa State.
*Corresponding author: [email protected]; 08124215650
Abstract
This research was aimed at identifying Staphylococcus aureus isolated from jollof rice samples
at selected cafeterias in Federal University Dutse campus. A total of fifteen (15) samples were
collected and processed within an hour in sterile containers from five (5) different cafeterias
(A, B, C, D and E). The total aerobic bacterial counts recorded from samples A, B, C, D and E
were 1.6 x 105, 1.1 x 105, 1.1 x 107, 9.3 x 104, 1.3 x107 cfu/g respectively while total
Staphylococcal counts recorded from samples A, B, C, D and E were 1.3 x 104, 6.5 x 103, 6.0 x
105, 4.0 x 103, 1.5 x108 cfu/g respectively. Based on staphylococcal counts recorded, sample A
result is unacceptable while samples B and D are within tolerable microbiological quality while
that of samples C and E are also of poor quality. Staphylococcus aureus isolated from the
samples was higher in samples C and E than in samples A, B and D. The isolate was susceptible
to Ofloxacin and Gentamicin based on susceptibility tests conducted. Closer supervision of food
handlers preparing and serving the food, most be carried out routinely by relevant authorities
within the campus with a view to preventing possible outbreak of food borne illness caused by
S. aureus.
Keywords: Staphylococcus aureus, jollof rice, bacterial counts, cafeterias
INTRODUCTION toxins and is found commonly on the skin, in
Consumption of safe foods should be a basic the nose and throats of up to 25% of healthy
human right despite the fact that numerous people as a normal flora. Cases of most S.
food items are often contaminated with aureus food borne illnesses reported in the
naturally occurring pathogenic microorganisms. literature are mostly triggered by poor hygiene
Greig et al. (2007) did estimate that about 2.5 exhibited by food handlers and grossly improper
billion people patronize food vendors in the food handling practices.
world over. According to Ogunyemi et al. Jollof rice is a sweet-smelling dish that is
(2015), ready-to-eat food items can potentially illustrious across the sub-region of West Africa
serve as a reservoir of pathogenic owing to its exceptional taste and subtle
microorganisms that have got the ability of spiciness (Adam, 2017). This author further
transmitting diseases. It has been reported by reported that Jollof is thought to have sprung
Itoandon et al. (2011), that the presence of up from the Senegambia region of West Africa
mesophilic microorganisms in food items is a amongst the indigenous Wolof people that refer
suggestion that pathogenic microorganisms are to it as benachin. Due to the popularity of
probably present in such food items. Some jollof rice for its irresistible taste and reported
authors (Balaban and Rasooly, 2000; Oranusi et cases of jollof rice being prone to
al., 2006a, 2006b) have reported a number of contamination with pathogens, this research
food items locally sold in Nigeria as vastly was conducted with a view to screening the
contaminated with Staphylococcus sp. jollof rice sold on the campus of Federal
Staphylococcus aureus has been reported over University Dutse for the possible presence of S.
the years as having the ability of causing food aureus.
poisoning. This submission has been supported
by the report of Monday et al. (2014) that the MATERIALS AND METHODS
bacterium can cause food poisoning and other Study Area
food-borne diseases. The study was conducted in designated area
In any community, Staphylococcus aureus has meant for commercial activities popularly
been considered as the major pathogen that called backside or bacteria (students’ version)
has got the ability of colonising and infecting situated on the campus of Federal University
both hospitalized patients exhibiting decreased Dutse North western, Nigeria. According to Peel
immunity and healthy immuno-competent et al. (2007), Dutse is the capital city of Jigawa
people. According to Centre for Disease Control state. The authors reported that Dutse lies on
and Prevention (2006), this bacterium produces latitude of 11o42’8.46” N and longitude of
9o20’2.4p6” E.
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 UJMR, Volume 4 Number 2, December, 2019, 43 - 48 ISSN: 2616 – 0668
Collection of samples 48 hrs. The procedure was carried out in
Jollof rice samples were purposively duplicate plates. At the end of the
collected from five (5) cafeterias (A, B, C, D incubation period, the different culture
and E) on Federal University Dutse campus, plates were examined for microbial growth
Jigawa State. Fifteen (15) different samples and the colonies were counted using the
(A1, A2, A3, B1, B2, B3, C1, C2, C3, D1, D2, D3, colony counter. The count for each plate was
E1, E2 and E3) of jollof rice comprising of expressed as colony forming unit per gram of
three (3) each from the restaurants were the sample (cfu/g) as prescribed by
collected within the period of four (4) weeks. Cheesbrough (2006).
All the samples were collected in sterile Isolation of Staphylococcus aureus
containers and transported within one (1) From the stock solution, 0.1mL was spread
hour of collection to the Microbiology onto Mannitol Salt agar plates with a sterile
laboratory for onward microbiological bent glass rod. The inoculated plates were
examination. incubated at 37oC for 48 hours. After
Media preparation incubation, the plates were observed for
Peptone water, Nutrient agar, Mannitol Salt characteristic colonies of S. aureus on the
agar and Muller-Hinton agar were employed Mannitol Salt Agar and were sub-cultured on
in this research and prepared according to nutrient agar slants and incubated at 37oC for
the instructions of the manufacturers. 24 hours. The slants were stored for Gram
Viable counts staining and biochemical characterization at
As prescribed by Cheesbrough (2006), ten 4oC as presented by Monday et al. (2014).
(10) grams of jollof rice for each sample was Gram staining
weighed aseptically using a weighing balance Gram staining was done according to the
and was placed into a sterile blender and procedure prescribed by Olutiola et al.
homogenised with 90mL Peptone water. (2000) with a view to identifying the isolate.
Further ten-fold serial dilutions of the Biochemical characterisation of
resultant homogenates were made to obtain Staphylococcus aureus
10-2, 10-3, 10-4 and 10-5 respectively. From 10- The choice of the various biochemical tests
4
and 10-5, 0.1mL was plated in replicate onto that were conducted with a view to
Nutrient agar using spread plate technique identifying Staphylococcus aureus isolated in
for total aerobic plate count. All inoculated this study was influenced by the prescription
plates were incubated at 37oC for 48 hours. of Barrow and Feltham (1993). Oxidase,
At the end of the incubation period, the Voges Proskauer, Coagulase, Lactose,
different culture plates were examined for Maltose, Mannitol, Fructose, Sucrose, Xylose,
microbial growth and colonies that developed Cellobiose, Phosphatase, Mannose, Nitrate,
were counted using the colony counter. The Arginine and Protease tests were conducted
count for each plate was expressed as colony according to the procedures prescribed by
forming unit per gram of the sample (cfu/g). Cheesebrough (2006); Ochei and Kolhatkar
Discrete colonies resembling Bacillus cereus (2008); Wilson (2012); Hemraj et al. (2013);
on Nutrient agar plates were sub-cultured Microbeonline (2019).
onto Nutrient agar slants and incubated at Antibiotic susceptibility tests
37oC for 24 hours. The slants were stored for Inocula of the test organisms were prepared
Gram staining and biochemical by taking and emulsifying distinct colonies
characterization as prescribed by Monday et from Nutrient agar on a sterile normal saline.
al. (2014). Antibiotic susceptibility testing was
Staphylococcal counts performed on all the isolates according to
Ten (10) grams of jollof rice for each sample the criteria of the Clinical and Laboratory
was weighed aseptically using a weighing Standard Institute (CLSI). A fresh sterile
balance and was placed into a sterile blender cotton-tipped swab was dipped into the
and homogenised with 90mL Peptone water. suspension and the excess liquid was
Further ten-fold serial dilutions of the removed from the swab by pressing it against
resultant homogenates were made to obtain the side of the tube. The surface of the
10-2, 10-3, 10-4 and 10-5 respectively. From all entire Muller Hinton agar plate was
the dilutions, 0.1mL was aseptically pipetted inoculated with the swab by streaking back
onto Mannitol Salt agar using spread plate and forth from edge to edge and was evenly
technique for total staphylococcal count. All distributed on the plate. The plates were
inoculated plates were incubated at 37oC for allowed to stand for few minutes, then
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multiple antibiotic disks were impregnated millimeter using a ruler placed at the reverse
using sterile forceps and then evenly side of the plates and compared to a
dispensed onto the agar surface. It was then standard interpretation chart used to
incubated at 37oC for 24 hours. categorize the isolate as susceptible,
Interpretation of results was done using the intermediately or resistant.
zone sizes. The zone was measured in
Statistical Analysis RESULTS
Data generated from the total viable and The results of the viable and staphylococcal
staphylococcal counts were analyzed using counts coupled with antibiotic sensitivity tests
Microsoft Excel. Descriptive statistics in form of conducted in this study are depicted in Tables 1
tables were employed to summarize the means and 2 respectively.
that were generated.

Table.1: Total Viable counts of jollof rice sold in the cafeterias on Federal university Dutse
campus.
Sample Cafeteria
Mean count for Mean count for Mean count for Mean count for Mean count for
cafeteria A cafeteria B cafeteria C cafeteria D cafeteria E
(cfu/g) (cfu/g) (cfu/g) (cfu/g) (cfu/g)
5 5 7 4
1 1.5×10 1.1×10 1.5×10 8.0×10 TNTBC
2 2.0×105 TNTBC 7.5×106 9.0×104 1.1×107
3 1.3×105 TNTBC TNTBC 1.1×105 1.5×107
MEAN 1.6×105 1.1×105 1.1×107 9.3×104 1.3×107
KEY: TNTBC= Too numerous to be counted
3 4 5 6
Standard = ≤10 cfu/g is acceptable; Between 10 - 10 cfu/g is tolerable; ≥10 cfu/g and above is
unacceptable (ICMSF, 2011)

Table 2: Total Staphylococcal count (cfu/g) of jollof rice sold in cafeteria in Federal university
Dutse.
Sample Cafeterias
Mean count for Mean count for Mean count for Mean count for Mean count for
cafeteria A cafeteria B cafeteria C cafeteria D cafeteria E
(cfu/g) (cfu/g) (cfu/g) (cfu/g) (cfu/g)
3 5
1 NG 5.0×10 6.0×10 NG 1.1×105
2 1.0×104 8.0×103 NG NG 1.5×105
3 1.5×104 NG NG 4.0×103 2.0×105
MEAN 1.3×104 6.5×103 6×105 4×103 1.5×108
Key: NG= No growth
2 3 4
Standard = ≤102cfu/g is satisfactory; Between 10 - 10 cfu/g is Marginal; 10 cfu/g is unacceptable;
5
≥10 cfu/g is potentially hazardous (ICMSF, 2011)

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 UJMR, Volume 4 Number 1, June, 2019 ISSN: 2616 - 0668

Table 3: Biochemical Characterization of the Isolate

Biochemical Tests Status
Oxidase _
Voges Proskauer +
Coagulase +
Lactose +
Maltose +
Mannitol +
Fructose +
Sucrose +
Xylose _
Cellobiose _
Mannose +
Phosphatase +
Nitrate +
Arginine +
Protease +
Identity of possible bacterium Staphylococcus aureus
Key: + = Positive; - = Negative

Table 4: Antibiotic Susceptibility Pattern of Staphylococcus aureus isolated from jollof rice sold
in cafeteria in Federal University Dutse.
Antibiotics Pattern of sensitivity
Sensitive Intermediate Resistance
Ofloxacin (5µg) 15mm _ _
Augmentin (30µg) _ _ 12mm
Ceftazidime (30µg) _ _ 10mm
Cefuroxime (30µg) _ _ 13mm
Gentamicin (10µg) 17mm _ _
Ceftriaxone (30µg) _ _ 23mm
Erythromycin (15µg) _ 15mm _
Cloxacillin (15µg) _ 8mm _

DISCUSSION contamination in the jollof rice sold at the

In this study, the mean viable plate count cafeterias (Table 2). Viable counts of
across all the cafeterias ranged between 1.1 bacteria in the jollof rice from cafeteria C
x 105 and 9.5 x 104 cfu/g (Table 1) while and E (1.1 x 107 and 1.3 x 107 cfu/g) were
mean staphylococcal count obtained ranged higher than that of jolly rice in cafeteria A, B
between 1.3 x 104 and 6.5 x 103 cfu/g (Table and D (1.6 x 105,1.1 x 105 and 9.3 x 104
2). As recorded in this study, a similar study cfu/g) respectively (Table 1).
conducted by Oranusi et al. (2013); Monday The staphylococcal counts of jollof rice sold
et al., (2014) reported high bacterial in cafeteria C and E (6.0 x 105 and 1.5 x 105
population in fried jollof rice examined in cfu/g) were higher than that of the jollof
their respective studies. The findings of this rice in cafeteria A, B and D (1.3 x 104, 6.5 x
study revealed that there was staphylococcal 103 and 1.5 x 105 cfu/g) respectively (Table

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2). The occurrence of Staphylococcus aureus resistant to Augmentin, Ceftazidime,

in jollof rice samples sold in cafeteria C and Cefuroxime, Ceftriaxone, Cloxacillin (Table
E was higher than that of cafeteria A, B and 4). These results are similar to the report of
E. These trends can be attributed to a study conducted by Akpomie et al. (2013)
variations in hygienic practices put up by the in which Staphylococcus aureus was sensitive
different cafeterias during this study to Ofloxacin, Gentamicin and Ciprofloxacin
The isolation of Staphylococcus aureus in this but resistant to Cefuroxime, Cefixime and
study is in agreement with the findings of Ceftriazone (Cephalosporin’s), Augmentin (β-
Taulo et al. (2009); Oranusi et al. (2013) in lactam inhibitor) and septrin
which this bacterium was implicated in (Sulphonamide).
contaminating ready-to-eat foods analysed in
their respective studies. The presence of CONCLUSION AND RECOMMENDATIONS
Staphylococcus aureus in food items is The results obtained in this study indicate
basically as a result of human contact and that the jollof rice consumed by members of
this suggests poor hygiene practices of the the University community that patronize
vendors since the bacterium is a normal flora these cafeterias is not safe for human
of the skin and nasal passage. Interestingly,45 consumption as samples from majority of the
Bibek (2001); Ali et al. (2008) have reported food vendors recorded staphylococcal counts.
that poor personal hygiene and inappropriate This, however, implies the deplorable state
food handling techniques are factors that of poor hygienic and sanitary practices
influence the chances of S. aureus being employed in the processing and handling of
transferred to jollof rice. Some authors foods prepared and served to the students
(Garret, 2002; Nichols et al., 2005) have that patronise the cafeterias. Therefore,
reported that Staphylococcus aureus effective sanitization of cooking areas and
produces a wide variety of toxins including food serving surfaces should be carried out to
staphylococcal entero-toxins that have got prevent cross contamination from utensils
enteric activity. These widely reported and flies. Utensils and surfaces should be
entero-toxins must have influenced the washed before and after use with hot, soapy
contamination of the jollof rice sampled and water or preferably, they should be sanitized
analysed in this study. with diluted bleach. Also a closer and routine
In the antibiotic sensitivity tests conducted in supervision of food handlers most especially
this study, Staphylococcus aureus was found those selling ready-to-eat foods, should be
to be susceptible to Ofloxacin and carried out by relevant authorities within the
Gentamicin (Table 4). It was intermediate to school campus to prevent possible outbreak
Erythromycin and also resistant, while it was of food borne illness.

Journal of Bangladesh Agric. Univ.,

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