Salmonella infection remains a serious problem worldwide and cause substantial economic loss, resulting in mortality, morbidity and poor growth. A total of 150 samples (50 each from Stool, Urine and Blood) were collected from 50 patients. from January to June 2015. The research was conducted in Microbiology laboratory of Kebbi State University of Science and Technology Aliero. The culture media used for this study were selenite broth, cysteine lactose electrolyte deficient agar, hektoen enteric agar, cooked meat agar, nutrient agar, chocolate agar and blood agar. The isolates were identified base on colonial appearance, gram staining reaction, microscopic morphology and series of biochemical tests. Blood sample yielded the highest number of salmonella isolates of 12 (24%), followed by stool sample with number of occurrence of 8 (16%), and urine sample yielded the lowest number of Salmonella spp isolated i.e. 2(4%). The results obtained also showed that 22 (21.8%) of the isolates were Salmonella species whereas 52 (51.5%), 25 (24.8%) and 2 (2.0%) were Escherichia coli, Staphylococcus aureus, and Shigella spp respectively. Out of 22 Salmonella species isolated, 13 (59.1%) were Salmonella. typhi, 6 (27.3%) were Salmonella paratyphi and 3(13.6%) were other Salmonella Species. Salmonella typhi and paratyphi are more prevalence compare to other non-typhoidal Salmonella. Simply because S. typhi and paratyphi A are species specific to humans and do not occur in other animals. From this research, it was concluded that blood culture is the best test samples for isolation of Salmonella followed by stool sample; this simply implies that typhoidal fever is higher than the nontyphoidal fever in this population.

1. Comparative evaluation of stool, urine and blood culture for isolation of Salmonella spp. from patients with clinical evidence of salmonellosis
WJM
Comparative evaluation of stool, urine and blood culture
for isolation of Salmonella spp. from patients with
clinical evidence of salmonellosis
1Yusuf, AB., 2Manga, SS., 1Kalgo, ZM.,1Aliyu B., 2Adesina, MA.
1
Department of Microbiology, Federal University, Birnin Kebbi, Kebbi State, Nigeria.
2
Department of Biological Sciences, Kebbi State University of Science and Technology Aliero, Kebbi State,
Nigeria.
Salmonella infection remains a serious problem worldwide and cause substantial economic loss,
resulting in mortality, morbidity and poor growth. A total of 150 samples (50 each from Stool,
Urine and Blood) were collected from 50 patients. from January to June 2015. The research was
conducted in Microbiology laboratory of Kebbi State University of Science and Technology Aliero.
The culture media used for this study were selenite broth, cysteine lactose electrolyte deficient
agar, hektoen enteric agar, cooked meat agar, nutrient agar, chocolate agar and blood agar. The
isolates were identified base on colonial appearance, gram staining reaction, microscopic
morphology and series of biochemical tests. Blood sample yielded the highest number of
salmonella isolates of 12 (24%), followed by stool sample with number of occurrence of 8 (16%),
and urine sample yielded the lowest number of Salmonella spp isolated i.e. 2(4%). The results
obtained also showed that 22 (21.8%) of the isolates were Salmonella species whereas 52 (51.5%),
25 (24.8%) and 2 (2.0%) were Escherichia coli, Staphylococcus aureus, and Shigella spp
respectively. Out of 22 Salmonella species isolated, 13 (59.1%) were Salmonella. typhi, 6 (27.3%)
were Salmonella paratyphi and 3(13.6%) were other Salmonella Species. Salmonella typhi and
paratyphi are more prevalence compare to other non-typhoidal Salmonella. Simply because S.
typhi and paratyphi A are species specific to humans and do not occur in other animals. From
this research, it was concluded that blood culture is the best test samples for isolation of
Salmonella followed by stool sample; this simply implies that typhoidal fever is higher than the
nontyphoidal fever in this population.
Keywords: Blood, Salmonella spp., Stool, Urine.
INTRODUCTION
Salmonella are gram-negative motile bacilli. The genus
Salmonella, which belongs to the family
Enterobacteriaceae, was named after Daniel E. Salmon,
an American veterinarian who first isolated Salmonella
choleraesuis from pigs with hog cholera in 1884 Kim et al.
(2004) .Salmonellae are ubiquitous human and animal
pathogens. They colonize virtually all animals including
poultry, birds, livestock, reptiles, rodents, domesticated
animals, and humans Subhash (2001) .As with the closely
related bacterium Escherichia coli, salmonellae are
potential enteric pathogens and a leading cause of
bacterial foodborne illness. In addition, Salmonella species
have been implicated in a spectrum of other diseases,
including enteric or typhoid fever (primarily Salmonella
typhi and Salmonella paratyphi), bacteria, endovascular
infections, focal infections (eg, osteomyelitis), and
enterocolitis (typically Salmonella typhimurium,
Salmonella enteritidis, and Salmonella Heidelberg) Kim et
al. (2004).
*Corresponding author: Kalgo Zaharaddin Muhammad,
Department of Microbiology, Federal University Birnin
Kebbi, Kebbi State, Nigeria. Tel: +2348031367195 Email:
zaharaddin28@gmail.com
World Journal of Microbiology
Vol. 4(1), pp. 133-138, January, 2018. © www.premierpublishers.org, ISSN: XXXX-XXXX
Research Article

2. Comparative evaluation of stool, urine and blood culture for isolation of Salmonella spp. from patients with clinical evidence of salmonellosis
Kalgo et al. 134
This result in substantial economic loss, mortality,
morbidity, poor growth and represents a serious problem
to food industries Khan (2007). Human spreads
Salmonella mainly through the stool. Food and water
borne transmission can occur when food and water are
contaminated with stool or through direct fecal-oral route.
Salmonellosis is a worldwide issue and People at risk of
serious complications due to Salmonella food poisoning
include: adults, pregnant women, infants, children, and
people who have compromised immune systems
Merchant and Packer, (1967). Symptoms usually include:
headache nausea, vomiting, fatigue, gastroenteritis,
abdominal cramps and bloody diarrhea with mucus and
sometimes reactive arthritis (Reiters Syndrome) Dworkin
et al. (2001).
The incidence of salmonellosis has markedly increased in
many countries; however, a paucity of good surveillance
data exists. In 2000, approximately 21.6 million worldwide
cases of typhoid fever caused 216,500 deaths Crump et
al. (2004).The incidence of typhoid fever in south-central
Asia, Southeast Asia, and, possibly, southern Africa was
high (>100 cases per 100,000 population per year). The
rest of Asia, Africa, Latin America, and Oceania (except for
Australia and New Zealand) typically see intermediate
rates of typhoid fever (10-100 cases per 100,000
population), while the incidence is low in the other parts of
the world (< 10 cases per 100,000 population). In countries
where typhoid fever is endemic, most cases of the disease
occur in children aged 5-19 years and young adults Bhan
et al. (2005).The incidence of Salmonella infections in the
United States has been stable since 2004 but has
decreased approximately 8% from 1996-1998 levels (8) In
2007, the reported annual incidence of salmonellosis was
14.9 cases per 100,000 population CDC (2007).The true
annual burden of nontyphoidal Salmonella infection in the
United States is calculated to be 520 cases per 100,000
population, compared with 13.4 laboratory-confirmed
cases per 100,000 population per year. This reflects an
estimate of approximately 38.6 cases of nontyphoidal
Salmonella infection for each culture-confirmed case
Voetsch et al.. (2004).
Typhoid fever is among the major widespread diseases
affecting the population in Nigeria and has been rated
eighth among these common infections Odugbemi et al.
(1994). Nigeria, like many other tropical and developing
countries, has been described as an endemic zone for
typhoid fever (Oboegbulam et al. (1995) and Idoko et al.
(1988)) In Nigeria, transmission of typhoid fever occurs all
year round but rates are slightly higher in April and July,
coinciding with the height of the hot, dry season and the
onset of the rainy season, respectively Oboegbulam et al.
(1995) ) . The highest number of cases of typhoid fever are
recorded during the rainy season in south-east Nigeria
Odugbemi et al. (1994). Typhoid fever has been reported
in all age groups and classes in Nigeria Rasaily et al.
(1994). Owing to the irregular nature of bacterial shedding,
several samples should be examined in order to identify
carriers Smith et al. (2008).Active surveillance on
salmonellosis in recent years is rare. Besides, there is a
paucity of data on studies emphasizing on isolation and
characterization of Salmonella species considering human
stool, blood and urine in Nigeria. A lot will be achieved
through the evaluation of these samples to ascertain which
of them will be the best to isolate these bacteria in order to
inform the clinicians on which sample to send for
diagnosis. Therefore, this research was aimed at isolating
and characterizing Salmonella species from patients
attending Federal Medical Centre B/ Kebbi.
MATERALS AND METHODS
Study population
The study population, were both male and female adult
patients who attended the General out patients
Department of Federal Medical Centre Birnin Kebbi,
Nigeria, with the clinical evidence of Salmonellosis (i.e.
headache nausea, vomiting, fatigue, gastroenteritis,
abdominal cramps and bloody diarrhoea with mucus)
Dworkin et al. (2001) , as diagnosed by the attending
physicians. These patients did not include those who were
on antibiotics about two weeks prior to specimen’s
collection.
Ethical clearance
Ethical approval was sought for from the Kebbi State
Ministry of Health. Informed consents were also obtained
from all the participants.
Sample collection
Three different samples (blood, stool, urine) were collected
from 50 patients attending Federal Medical Center Birnin
Kebbi with clinical evidence of salmonellosis i.e a total of
150 samples were obtained. All patents were instructed on
how to collect appropriate specimens. Blood samples were
collected from patients using blood sample bottles by the
attending clinician while stool and urine specimens were
collected using a sterile, clean, dry, disinfectant-free, wide-
necked container with tight fitting lid. All the specimens
were taken to the laboratory for analysis without delay.
Bacteriological investigation
Stool culture
The purudent or mucoid parts of the stool samples were
picked using a sterile wire loop and inoculated into selenile
broth medium and then incubated at 370C overnight. (18-
24hrs). The broth culture was then subculture into
salmonella shigella medium agar, and hektoen enteric
agar, the plates were then incubated at 370C for 24hrs.

3. Comparative evaluation of stool, urine and blood culture for isolation of Salmonella spp. from patients with clinical evidence of salmonellosis
World J. Microbiol. 135
Urine culture
Using a sterile wire loop, a loop full of the urine sample
was picked and inoculated into Bismuth sulfite agar,
McConkey agar, Blood Agar, XLD (xylose, lysine
deoxycholate agar). The plates were then incubated at
370C for 24hrs.
Blood culture
Using a sterile syringe and needle, 4ml of whole blood from
the patient was dispensed into 20ml of cooked meat
medium. The mixture was then incubated at 370C for 24-
48hrs before subculture into blood agar, CLED, hektoen
enteric agar and salmonella shigella agar. The plates were
then incubated at 370C for 24hrs. The sub culturing was
done three times before conclusion that there is no growth.
Identification of isolates
The suspected colonies were characterized
morphologically using Gram’s staining method according
to the method described by Merchant and Packer, (1967)
. For carbohydrate fermentation tests, triple sugar iron
agar (TSIA) slant reaction, methyl red-Voges-Proskauer
(MR-VP) and Indole reaction tests were carried out for
identification of suspected Salmonella according to the
methods described by (Douglas et al. (1998) and OIE
(2000)).
RESULTS
Table 1: Salmonella isolates obtained from three
different samples
Sample Source No Tested No. of Salmonella
Stool sample 50 8(16%)
Urine sample 50 2(4%)
Blood sample 50 12(24%)
150 22(14.7%)
Table 2: Prevalence of Salmonella species and other
bacteria in three different samples
Isolates Stool Urine Blood Total Percentage
(%)
E. Coli 21 30 1 52 51.5%
Staph. aureus 0 10 15 25 24.80%
Shigella 2 0 0 2 2.0%
Salmonella 8 2 12 22 21.8%
Total 35 42 24 101 67.3%
Total number of salmonella isolates obtained each sample
source.
Table 3: prevalence of Salmonella species in three different samples
Salm. Species Stool Urine Blood Total Percentage (%)
S. typhi 5(38.5%) 2(15.4%) 6 (46.2%) 13 59.1%
S. paratyphi A 2(33.3%) 0(0%) 4(66.7%) 6 27.3%
Other salmonella SPP 2(66.7%) 0(0%) 1(33.3%) 3 13.6%)
Total _ _ _ 22 100%
Figure 1: Pie chart showing the percentage of different Species of Salmonella

4. Comparative evaluation of stool, urine and blood culture for isolation of Salmonella spp. from patients with clinical evidence of salmonellosis
Kalgo et al. 136
Figure 2: Bar chart showing the number of occurrence of Salmonella spp from various samples.
DISCUSSION
Salmonella causing human disease are divided into
human-restricted typhoidal serovars (Typhi and Paratyphi)
causing typhoid fever, and non-typhoidal Salmonella
(NTS) serovars which have a broader host-range and are
frequently zoonotic Han et al. (2011) .This study verified
that out of 150 specimens (50 each from blood, urine and
stool) analysed, 22 were positive for salmonella spp giving
the overall prevalence of 14.7%. It was also observed from
this study that blood sample yielded the highest number of
salmonella isolates of 24%, followed by stool sample, but
it was documented from this study that urine sample
yielded the lowest percentage of Salmonella spp isolated.
This result is in conformity with the work of (Melita (2011)
and Kabir et al. (2007)) who also reported high positive
results of salmonella from blood samples. From the result
outlined above, it is clear that incidences of typhoid fever
are relatively high, although less; when compared to some
instances reported in Nigeria (Melita (2011) and Kabir et
al. (2007)).
The results of the study indicated that the frequency of
isolation of S. Typhi were 6 (46.2%) from blood sample, 5
(38.5%) were from stool sample and 2 (15.4%) were from
urine samples. S. paratyphi A has the frequency of 4
(66.7%) from blood samples, 2 (33.3%) were from stool
samples and none of the salmonella paratyphi A were
isolated from urine samples. This result is in conformity
with the study conducted by Okonko et al. (2010) where
salmonella typhi and Salmonella paratyphi A has the
highest occurrence from blood samples. This study also
agrees with the work of Okonko et al. (2010) where they
documented only few number of salmonella spp from stool
samples than the blood samples.
The importance of Salmonellosis in public health sector is
a growing concern day by day throughout the world and
over the last several decades there have been significant
shift in predominant Salmonella serovars associated with
human infections Isa et al. (2013). Salmonellosis in the
past has caused tremendous loss to society inmany
countries around the world. Two to four million of cases
have been reported annually and yet a significant number
of cases have been unreported worldwide. Non-typhoidal
Salmonella is the leading cause of food borne illness and
its increasing antimicrobial resistance is associated with
higher risks of hospitalization Steven et al. (2011).
Out of the 22 patients Salmonella species isolated, 8 of
them were actually having typhoid fever because the
organisms (Salmonella) were isolated from both, the blood
and stool sample of these patients, since their blood
culture were positive. Isolation of Salmonella from blood is
definitive proof of septicemia Han et al. (2011) . The
remaining four patients who are positive to Salmonella
from their stool sample but negative to their blood culture
samples may be carriers since the isolation of the
organisms from faeces is not as valuable as blood culture
in diagnosis Han et al. (2011) .
CONCLUSION
From the above results of this study, it was concluded that
blood samples yielded highest number of Salmonella spp
when compare to stool and urine samples, this simply
implies that, most of the patients are suffering from
septicaemia caused by Salmonella spp. The results
obtained also showed that out of 22 Salmonella species
0
2
4
6
8
10
12
14
Blood Stool Urine

5. Comparative evaluation of stool, urine and blood culture for isolation of Salmonella spp. from patients with clinical evidence of salmonellosis
World J. Microbiol. 137
isolated, Salmonella typhi has the highest number of
occurrence of 13 (59.1%), followed by Salmonella
paratyphi and other Salmonella spp with the number of
occurrence of 6 (27.3%) and (13.6%) respectively.
Therefore, blood culture should be of priority for the
diagnosis of the disease cause by Salmonella spp in this
population.
RECOMMENDATION
1. The clinician (Medical Doctors) should be educated on
the importance of confirmation of Salmonella infection
before prescription of antibiotic in order to avoid the
emergent of resistance strain to available sensitive drugs.
2. Further studies should increase the number of patients
for the research and more samples apart from the one
used should be included in future studies.
3. PCR and other modern identification procedures should
be used for proper identification of the species and strain
of Salmonella.
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6. Comparative evaluation of stool, urine and blood culture for isolation of Salmonella spp. from patients with clinical evidence of salmonellosis
Kalgo et al. 138
Accepted 18 December, 2017.
Citation: Yusuf AB, Manga SS, Kalgo ZM, Aliyu
B,2Adesin, MA (2018). Comparative evaluation of stool,
urine and blood culture for isolation of Salmonella spp.
from patients with clinical evidence of salmonellosis.
International Journal of Cardiology and Cardiovascular
Research 4(1): 133-138.
Copyright: © 2018 Kalgo et al. This is an open-access
article distributed under the terms of the Creative
Commons Attribution License, which permits unrestricted
use, distribution, and reproduction in any medium,
provided the original author and source are cited.