Volume 9, Issue 11, November – 2024 International Journal of Innovative Science and Research Technology

ISSN No:-2456-2165

Prevalence of Salmonella Species Infection and

Antibiotics Susceptibility of Isolates in
Day-Old-Chicks from Hatcheries and
Farm Markets in Ibadan, Nigeria
Dr. Alaba Opeoluwa A1; Olayinka Ishola O.2; Dr Okunade3

Abstract:- Salmonellae is a prevalent infection in the All the six Salmonellae isolates used for the
chicken industry. The infection in poultry is a persistent antibiotics susceptibility test showed 100% resistance to
global problem that poses a hazard to both animal and Ampliclox, Amoxicillin clavulanate (Augmentin),
public health. Salmonellae leads to reduced production Cefotaxime, Imipenem and Nalidixic acid. The isolates
efficiency and mortality in poultry, resulting in significant had moderate resistance to Ofloxacin (83%),
economic losses worldwide. In order to ensure the health Nitrafuratoin (83%), Ceftriaxine (67%) and Levofloxacin
and productivity of poultry, it is essential to begin with (67%). The Salmonellae isolates were found to be only
high-quality day old chicks that are free from diseases. 50% susceptible to Cefexime, 33% susceptible to
Poultry meat and eggs are significant contributors to the Gentamycin, 17% susceptible each to Cefuroxime and
animal protein supply in Nigeria. A significant number of Nitrafuratoin while all the isolates were not susceptible
newly hatched chicks utilized for production are mostly (0%) at all to the remaining eight antibiotics.
sourced from the south-western states of Ogun and Oyo,
and then moved to other regions within the country. This Salmonellae infections of day old chicks sold in
study aimed to examine the prevalence of Salmonellae major markets in Ibadan was confirmed in this study. The
infections and the antibiotic susceptibility pattern of isolates were multi-drug resistant and portending
isolates from day-old chicks produced in hatcheries and challenge to treatment as well as public health risks.
marketed in major markets in Ibadan, Oyo states, There is a need for regular Salmonellae surveillance of
Nigeria. day old chicks in farm markets and hatcheries, increased
awareness programs for farmers and hatchery operators
This study was conducted using a cross-sectional by government on proper biosecurity measures and risks
design and focused on key farm markets where day-old involved in indiscriminate use of antibiotics in poultry
chicks are sold, as well as three hatcheries that were and the resultant effects of antimicrobial resistance on
carefully selected. Farm markets and their respective animal and human health.
locations, including the local government regions, were
identified in Ibadan. Keywords:- Salmonellae, Day-Old Chicks, Hatchery and
Anti-Microbial Resistance.
A total of 135 cloacal swabs were collected from
randomly selected day-old chicks. These chicks were I. INTRODUCTION
obtained from 10 sellers at three major agricultural
markets in three Local Government Areas (LGAs) and Salmonellae infection in chicken is a persistent global
three hatcheries. Salmonellae isolation was performed problem that poses a hazard to both animal and public health
using culture techniques that involved buffered peptone (Leon, 2015).The reason for its prevalence in the food sector
water, Rappaport-Vassiliadis broth, and Xylose Lysine is its ability to endure and spread across many stages or
deoxycholate agar. The isolates were characterised using phases in chicken production. Salmonellae is a group of
standard biochemical testing and antibiotics bacteria belonging to the family Enterobacteriaceae. These
susceptibility determined using agar diffusion. bacteria are characterised by being Gram-negative, oxidase
negative, catalase positive, and having a rod-like shape
The overall prevalence of Salmonellae infection old without the ability to make spores. Additionally, they are
day old chicks produced by hatcheries and sold in major facultative anaerobes. The ideal temperature for the growth
farm markets in Ibadan is 4.4% (6/135). Day old chicks of Salmonellaee is 37°C. However, it has been observed to
from the farm markets in egbeda had the highest thrive at temperatures as low as 2-4°C and as high as 54°C.
prevalence of 10.0 % followed by Ibadan South West of Salmonellae is capable of surviving in a broad variety of pH
3.3% and Ibadan North Est of 0.0%. from the hatcheries, levels, ranging from a minimum of pH 3.8 to a maximum of
day old chicks from hatchery A has the highest prevalence pH 9.5. However, it thrives best in an environment with a pH
of 13.3% while hatchery B and C had 0.0%. between 6.5 and 7.5. A water activity (𝑎𝑤) below 0.94
prevents the growth of Salmonellae. However, at specific

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Volume 9, Issue 11, November – 2024 International Journal of Innovative Science and Research Technology
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temperatures, a low 𝑎𝑤 is thought to provide protection wash room, clean-room, and additional rooms specific to the
against Salmonellae (Ryan et al., 2017). Salmonellae is type of hatchery. These additional rooms may include a small
widely found in the environment and can cause a variety of office, lounge and lunch room, rest room, generator or tool
infections in both humans and animals. It occurs naturally in room, box storage room, and electrical control room (Tamil
the intestines of both domestic and wild animals, such as Nadu Agriculture University, 2019).
chickens, swine, cattle, birds, dogs, rats, tortoises, turtles, and
cats (Demirbilek, 2017). Ensuring proper hygiene is crucial when it comes to
designing and managing a hatchery. Optimal hatching rates
The agricultural sector continues to be the primary and chick quality necessitate this. The safeguarding against
driver of the Nigerian economy, responsible for more than the introduction of harmful substances into hatching eggs and
38% of non-oil foreign exchange revenues and employing newly hatched chicks or poults, especially in relation to
around 70% of the working population. The poultry sub- targeted disease prevention measures (such as Salmonellaee),
sector in Nigeria's agriculture is highly commercialised and is becoming more and more important in the functioning of
has significantly improved the lives of the less privileged hatcheries (Behoir, 2021). The hatchery is a significant
population through minimal investment and low-cost source of disease risk across the entire poultry production
technologies (Jibril et al., 2020). Every year, a total of 454 cycle (Deori et al., 2019). Inadequate biosecurity measures at
billion tonnes of meat and 3.8 million eggs are produced, hatcheries not only diminish the calibre and volume of newly
from a population of 180 million birds. Poultry meat and eggs hatched chicks, but also facilitate the transmission of disease-
are the primary sources of animal protein in Nigeria, as well causing agents, result in ecological contamination, and
as in several developing nations, due to their cost- impact the well-being of employees (FAO, 2015). Effective
effectiveness and widespread acceptance. Regrettably, the hatchery management encompasses The owner's living area
expansion of this significant agricultural subsector is severely should be separated from the hatchery set-up. Before entering
jeopardised by several infectious diseases, including those and after leaving the hatchery, it is necessary to change
induced by Salmonellae species (Jibril et al., 2020). footwear and wash hands. There should be a one-way
Salmonellosis, a significant food-borne illness worldwide, is movement and unidirectional flow of air. It is important to
projected to cause around 93.8 million cases of gastroenteritis have separation between the incubation, hatching, and bird
annually, resulting in 155,000 deaths (Majowicz et al., delivery areas. Fumigation of eggs, egg trays, egg setters, and
20102017). hatchers should be done using formaldehyde solutions or
potassium permanganate (Esatu et al., 2016). This process
The global public health community is greatly should take place in a properly equipped egg fumigation
concerned about the isolation of Salmonellae from food cabinet.
products and its antibiotic resistance (Ishola et al., 2021).
Research has demonstrated that Salmonellae serotypes Salmonellae is often spread in two ways: vertically
exhibit resistance to many drugs (Ziba et al., 2020). (from parent to offspring) or horizontally (between
Antimicrobial resistance can be attributed to either mutations individuals). The hatchery is a significant source of both
in genes found in the chromosomes (known as intrinsic vertical and horizontal transmission of Salmonellae
resistance) or the acquisition of resistance determinants (Liljebjelke et al., 2005). Transmission of Salmonellae from
through plasmids (known as extrinsic resistance). The parent to progeny birds through vertical transmission
primary cause of intrinsic resistance is selection pressure, (transovarian) has been observed for multiple Salmonellae
while extrinsic genes are obtained through horizontal transfer serotypes. Experimental evidence has demonstrated that
(Katiyar et al., 2020). Antimicrobial resistance (AMR) has Salmonellae infection can occur in developing eggs within
become a major concern in both human and animal medicine the reproductive canal, specifically in the ovary (unshelled
worldwide. It is now widely acknowledged as a substantial eggs) and shell glands of infected hens (Liljebjelke et al.,
and growing threat to global public health and food security 2005; Cardoso et al., 2021). Transovarian infection in the egg
(Wall et al., 2016). leads to subsequent infection in chicks or poults and is a
significant method of transmitting salmonellosis in chickens.
A hatchery is a controlled environment facility or Certain chickens that are infected become asymptomatic
building specifically designed for the artificial incubation of carriers and consistently pass on the infection to their
eggs (McMahon, 2022). A hatchery fulfils the crucial offspring. The method of disease transmission is very crucial
function of gathering hatching eggs from the breeder farm in hatcheries since it might lead to the extensive spread of the
and distributing recently hatched chicks to a commercial diseases. Kuria, 2023 Chicks that are infected by hens
poultry farm (Kim & Kim, 2016).The contemporary approach typically become carrier chickens, capable of spreading the
to constructing hatcheries aims to provide an optimal physical infection through hatching, handling, and transit.
setting that maximises the effectiveness of converting a Salmonellae bacteria reside within the gall bladder and
hatching egg into a viable and robust chick using artificial exhibit resistance to antibiotics, rendering them impervious
incubation and hatching techniques (Deori et al., 2019). The to eradication (Overzicht, 2022).
hatchery facility consists of multiple rooms dedicated to
particular functions. The rooms in a hatchery typically consist The transmission of Salmonellae horizontally occurs by
of a shower room, sorting room, fumigation room, egg the route of ingestion of faecal matter (Kuria, 2023). The
holding room, pre-incubation warming room, setter room, majority of adult chickens become sick by consuming foods
candling or dark room, hatcher room, chick holding room, that have been polluted. Salmonellae can be mechanically

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ISSN No:-2456-2165

transmitted by dogs, cats, rodents, and insects (Kuria, 2023). East, Ibadan South East and Egbeda Local government were
Additionally, Salmonellae can enter a day-old chick through randomly selected for the study and a total of three (3) farm
production materials like tools, egg trays, crates, and markets, one from each of the three LGAs, were purposefully
containers that have not been cleaned or disinfected recruited based on the presence of commercial chick vendors
(Overzicht, 2022). Additionally, transmission through consent. Each Farm market was visited twice with 15 samples
cannibalism and inhalation has been documented (Kuria, collected per visit and the hatcheries were visited once with
2023). 15 samples collected per visit. Three vendors were randomly
. selected from which three chicks were sampled using the
During vertical transmission, birds that are infected with systematic random sampling method per-vendor. Afterwards,
Salmonellae spp. From their mother to them internally and each vendor was asked further questions for more
they become carriers of the bacteria. This not only poses a information.
threat to the economic aspects of poultry production, but also
to public health in general. These birds appear to be in good Cloacal swab samples were collected by gently rotating
health and are charming, but they have the ability to spread the cotton swab within the cloacae, with care taken to avoid
Salmonellae to any animal or human they come into contact contact with other areas to prevent contamination. All
with. The Centre for Disease Control and Prevention (CDCP) samples were transported in sterile tubes on ice to the
in 2022 documented instances of Salmonellae transmission Veterinary Public Health Laboratory of the Department of
from newborn chicks to humans. Annually, instances of Veterinary Public Health and Preventive Medicine,
Salmonellae outbreaks arise when individuals come into University of Ibadan, for bacteriological analysis
touch with baby poultry harbouring the Salmonellae
bacterium. The source of this information is the Minnesota D. Study Population
Department of Health, specifically from the year 2022. The study population were day-old-chicks produced at
the hatcheries and sold by vendors at the selected markets.
II. METHODS
The vendors(n=30) were also used as the study
A. Study Area population to obtain information regarding the use of
The location for this study was Ibadan. Ibadan is the vaccines and or drugs on the day-old-chicks.
capital of Oyo State, Nigeria. Oyo State is an inland state in
South-western Nigeria, with coordinates of 7.377° N and E. Study Population
3.947° E. The study population were day-old-chicks produced at
the hatcheries and sold by vendors at the selected markets.
B. Study Design
The study was a cross sectional study involving day-old The vendors(n=30) were also used as the study
chicks produced by hatcheries sold at major Farm Markets population to obtain information regarding the use of
situated within three randomly selected local government vaccines and or drugs on the day-old-chicks.
area (LGA). The LGAs are Ibadan North East, Ibadan South
West and Egbeda local government area. One-hundred and F. Inclusion Criteria
Forty-five (145) samples were collected from vendors and Samples were obtained from chick vendors in major
hatcheries from randomly selected hatcheries and three major market locations within Ibadan North East LGA, Ibadan
markets within the LGAs. South East LGA and Egbeda LGA having chick markets.
Sample Size Samples were collected from live birds irrespective of the
source, breed, clinical status and vaccination status.
Sample size estimation for day old chicks was
determined using the formula; G. Exclusion Criteria
Vendors selling other poultry birds and chickens more
n = Z2Pq (Thrusfield,2005) than one-day old were excluded from the study. Dead chicks
2
d were also excluded from the samples.
Where Z= level of significance (1.96), P=Prevalence, q=1-P,
d = level of precision (5%) III. RESULTS

Z=1.96, P=5% (Ezeibe et al., 2019) d=0.05, A. Identification of Major farm Markets where Day-Old-
= (1.96)2*0.05*(1-0.05)/ (0.05)2 Chicks are Sold in Ibadan
= 3.8416*0.05*0.95/0.0025 The major farm markets where day-old-chicks are sold
=73 in Ibadan include Obasanjo farm market located at Oluyole,
n/1-f, where f =10% contingency Ibadan South-West Local Government Area (LGA), Provider
73/1-0.1=81 farm market located at Monatan, Ibadan North East LGA and
Easy farm market located at Ice-block along new Ife-road,
C. Sample Technique and Collection. Egbeda LGA, Ibadan.
A multi-stage sampling method was used to select
chicks for sampling. First, three hatcheries and three local
government areas (LGAs) in Ibadan namely Ibadan North

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 Isolation on Xylose Lysine Deoxycholate Agar  Result of Triple Sugar Iron (TSI) Agar Test for
All sample tested positive for Rappaport Vassiliadis Salmonellae
with Soya (RVS) broth as colour change was observed in the Eighty (80) isolates were tested with TSI agar out of
RVS from blue to colourless (Fig. 1). Isolation and which sixty-nine (69) tested positive due to glucose
identification were carried out using Xylose Lysine fermentation, non-lactose and or sucrose fermentation,
Deoxycholate agar. Colonies of varying sizes, small and positive result for gas production and positive result for
transparent with black centres and slightly raised were hydrogen sulphide (H2S) production ((Table 1; Figure 3);
observed (Fig. 2). Of all the 135 samples, sixty-nine (80) indicative of sulphite reaction by the production of black
were positive for Salmonellae on XLD agar (Table 1). Ferrous Sulphate as stated by (Lehman, 2016).

 Gram Staining
Upon viewing of microscopic slides at x100
Magnification, pinkish short rod shape bacilli cells were seen
which indicates a Gram-negative-bacteria.

Fig 1: a and b Showing Rappaport Vassiliadis before Incubation, Bluish Colour (a) and after 24 Hours Incubation with Change in
Colour to Colourless (b)

Fig 2: Result on XLD Agar

Fig 3: Result on Triple Sugar Iron test (TSI)

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Table 1: XLD agar, Gram Staining and Biochemical Characterisation Results for Salmonellae spp. Isolated from Day Old Chicks
Sold at Farm Markets and Hatcheries in Selected Local Government Areas in Ibadan
Location No. of XLD Gram TSI Catalase Indole Oxidase Sugar fermentation test
sample Agar Staining test test test
-ve +ve +ve +ve -ve Glu Malt Man Lac Suc
+ve +ve +ve -ve -ve
Ibadan 30 17 17 16 16 16 15 1 1 1 1 1
South East
Ibadan 30 15 15 15 15 15 11 0 0 0 0 0
North East
Egbeda 30 22 22 20 20 10 19 3 3 3 3 3
Hatchery A 15 11 11 8 8 8 6 2 2 2 2 2
Hatchery B 15 4 4 3 3 3 2 0 0 0 0 0
Hatchery C 15 11 11 9 9 9 7 0 0 0 0 0
Total 135 80 80 69 69 69 60 6 6 6 6 6

 Catalase Test
Presumptive isolates for Salmonellae were confirmed as
catalase positive for all isolates on TSI as it produced
effervescence after a colony of the isolate was added to
hydrogen peroxide using a microscopic slide (Table 1).

 Indole Test
The presumptive isolates for Salmonellae were
confirmed indole negative for all isolates on TSI as they did
not form red-rings at the meniscus of the overnight broth
culture (Figure 4)

 Oxidase Test
Sixty (60) isolates for Salmonellae on TSI were
confirmed oxidase negative as they do not change colour
within five (5) seconds on treated filter paper with oxidase
reagent (Table 1) Fig 4: Result of Indole Test

 Sugar Fermentation B. Prevalence of Salmonellae in day-old chicks

Out of the fifty (50) selected isolates for sugar test, six Out of a total of 135 cloacal swab of chicks screened for
(6) were confirmed with the colour change from red to yellow Salmonellae spp., six (6) were confirmed positive with the
for the fermentation of Glucose, Mannose and Maltose with overall prevalence of Salmonellae infection being 4.4%
no colour change for non-fermentation of Lactose and (6/135) (Table 2) (Figure 4). Total confirmed Salmonellae
Sucrose (Table 1). isolates from the farm market were four (4) with a prevalence
of 2.9% (4/135) while the confirmed isolates from the
hatcheries were two (2) with a prevalence of 1.5% (2/135)
(Table 2).

Table 2: Prevalence of Salmonellae Infection in Day-Old Chicks from Major Hatcheries and Local Government Areas (LGAs)
where the Major Farm Markets are Located in Ibadan
LGA/Hatchery No. of Samples No of Confirmed Salmonellae Prevalence
Collected Isolates
Ibadan South West 30 1 3.3%
Ibadan North East 30 0 0.0%
Egbeda LGA 30 3 10.0%
Sub-total 90 4 2.9%
Hatchery A 15 2 13.3%
Hatchery B 15 0 0.0%
Hatchery C 15 0 0.0%
Sub-total 45 2 1.5%
Total 135 6 4.4%

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Fig 5: Prevalence of Salmonellae Infection in Day-Old Chicks from the Local Government Areas (LGAs) where the Major Farm
Markets are Located in Ibadan

Fig 6: Prevalence of Salmonellae Infection in Day-Old Chicks from Major Hatcheries and the Local Government Areas (LGAs)
where the Major Farm Markets are Located in Ibadan

C. Antibiotics Susceptibility Pattern of Salmonellae spp. in Ceftriaxine (67%) and Levofloxacin (67%) (Tables 3 and 4)
Day-Old Chicks (Figure 9).
All the confirmed Salmonellae isolates (n = 6) subjected
to antibiotic susceptibility test showed 100% resistance to The Salmonellae isolates were found to be only 50%
Ampliclox, Amoxicillin clavulanate (Augmentin), susceptible to Cefexime, 33% susceptible to Gentamycin,
Cefotaxime, Imipenem and Nalidixic acid. The isolates had 17% susceptible each to Cefuroxime and Nitrafuratoin while
moderate resistance to Ofloxacin (83%), Nitrafuratoin (83%), all the isolates were not susceptible (0%) at all to the
remaining eight antibiotics (Tables 3 and 4) (Figure 9).

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Fig 7: Antibiotics Susceptibility Pattern of Salmonellae spp. with Arrow Showing zone of Inhibition

Table 3: Antibiotics Susceptibility Pattern

Isolates CRO ACX ZEM LBC CXM AUG CTX IMP OFX GN NA NF
/antibiotics
Q/2/NL I R R I R R R R I S R R
A/2/BL I R S R S R R R I S R 1
HA/14/NL I R I I R R R R R R R I
U/3/NL R R S 1 I R R R I R R I
R/1/NL R R S I R R R R I R R I
HA/2/BL I R R R R R R R I I R S

Table 4: Frequencies of Resistance, Intermediate and Susceptibility of Antibiotics Agent

S/N Antimicrobial Frequencies of Frequencies of Frequencies of
Agent Sensitivity (%) Intermediate (%) Resistance (%)
1 Ceftriaxone 0.0 67 33
2 Amplicox 0.0 0.0 100
3 Cefexime 50 17 33
4 Levofloxacin 0.0 67 50
5 Cefuroxime 17 17 67
6 Amoxycillin 0.0 0.0 100
7 Cefotaxime 0.0 0.0 100
8 Imipenem 0.0 0.0 100
9 Ofloxacin 0.0 83 17
10 Gentamycin 33 17 50
11 Nalidixic acid 0.0 0 100
12 Nitrafuratoin 17 83 17

Fig 8: Antibiotics Susceptibility Pattern of Salmonellae spp.

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D. Use of Vaccines and / or Antibiotics on Day-Old-Chicks further information about the chicks. Out of the thirty (30)
by Vendors at Farm Markets and Hatcheries in Ibadan vendors, eight (8) vendors were found to administer
Vendors (n=30) which (n=10) are from each LGA and drugs/antibiotics while 2 (two) vendors administered vaccine
Hatchery Operators (n=3) were asked some questions for (Table 5) (Figure 9)

Table 5: Use of Vaccines and / or Antibiotics on Day-Old-Chicks by Vendors at Farm Markets and Hatcheries in Ibadan
LGA/Hatchery Vaccine Drug/Antibiotics
Ibadan South West (n=10) 1 5
Ibadan North East (n=10) 0 1
Egbeda (n=10) 1 2
Hatchery A (n=1) 0 0
Hatchery B (n=1) 0 0
Hatchery C (n=1) 0 0
Total 2 8

Fig 9: Use of Vaccines and / or Antibiotics on Day-Old-Chicks by Vendors at Farm Markets and Hatcheries in Ibadan

IV. DISCUSSION (2019) who reported 5% prevalence of Salmonellae spp.in

day-old-chicks in Akwa-Ibom State in Nigeria and also Jibril
The presence of Salmonellae in day-old chicks is one of et al., (2023) who reported 10.7% prevalence of Salmonellae
the most important risk factors for the introduction of spp. in day-old-chicks in three (3) states of the northwest
Salmonellae into poultry farms. Salmonellae infection does Nigeria. Result of findings were lower than that of
not only decrease production performance, it also causes Albuquerque et al., (2014) who reported 0.58% prevalence of
death of poultry birds (Wang et al., 2020). In this study, only Salmonellae spp.in day-old-chicks in Brazil. Salmonellae
three (3) major farm markets where day-old-chicks are sold infection is still one of the most important problems of
in Ibadan which are located in Oluyole, Ibadan South-West poultry worldwide despite various prevention and control
LGA, Monatan, Ibadan North East LGA and Ice-block along measures including vaccine and drug use (Wang et al., 2020).
new Ife-road, Egbeda LGA, Ibadan were identified. Studies has shown that Salmonellae illness in humans have
Considering the importance of day-old-chicks to been linked to zoonotic transmission from day-old-chicks via
development of poultry industry, there is a need to have more poultry products to humans and day-old-chicks have
well–organised farm markets where day-old-chicks are sold previously been associated with multistate outbreaks of
in Ibadan. salmonellosis (Jibril et al., 2023).

In this study, out of the 135 day-old-chicks examined The results of this study showed that day-old-chicks
for the presence of Salmonellae species, 2.9% (4/135) from Hatchery A had the highest (13.3%) while the other two
prevalence were from the Farm markets and 1.5% (2/135) hatcheries had lowest (0.0%) prevalence of Salmonellae. This
from the hatcheries with an over-all prevalence of 4.4% could be due to better hygienic practices by Hatcheries B and
(6/135).This findings were lower than that of Ezeibe et al., C. Kuria (2023) reported that there can be transmission of

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Salmonellae from the hatching environment to the chicks.  Government should make and enforce laws to restrict
The higher (10.0%) prevalence of Salmonellae infection in non-licensed veterinary medical practitioners from using
day-old-chicks from the farm market located in Egbeda LGA antibiotics indiscriminately in poultry to prevent
compared to the other two farm markets within Ibadan South antimicrobial resistance.
West and Ibadan North East LGAs could probably be due to  Farmers and vendors need to be educated on the resultant
poor biosecurity by the vendors at Egbeda LGA. effect of irregular use of antibiotics on the birds at day-
old
This findings from the antimicrobial susceptibility test
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