ABSTRACT
Antimicrobial resistance (AMR) is a worldwide health concern and use of antimicrobials in animal production is a major cause of AMR in humans. This study was conducted to determine factors that influence choice and application of antibiotics by broiler farmers in Abia State-Nigeria by administration of structured questionnaires to 100 farms. The study also evaluated antimicrobial resistance profile of bacteria isolated from broiler-chicks in the Abia State by conducting antimicrobial sensitivity test to ten antimicrobials commonly used in poultry farming in Abia State and developed a treatment-strategy for prevention of AMR and for curing already resistant infections of salmonella paratyphoid by stabilizing Streptomycin with Medicinal Synthetic Aluminium Magnesium Silicate. All 56 poultry farms that responded to questionnaires administered to them accepted that they use antibiotics on their farms. 67% of the farmers in Abia State were males, with first degree (34.5%) small-scale farmers (46%), keeping broilers (70%) in deep litter system (70%). They said that drugs they use to treat their birds are as prescribed by Veterinary Doctors (76.4%). Drug-formulations containing tetracycline (90%) are the ones most of them use. 70.4% of the farmers had good knowledge of laws regulating use of antibiotics in raising food animals. They also know the problem of Antimicrobial resistance. Of a total of 180 samples tested, 21% yielded presumptive Salmonella isolates in cultures with the two commonly encountered zoonoses, S. typhimurium and S. enteritidis respectively, accounting for 2.63% (1) and 97.36% (37) isolates of the Salmonella isolates when serotyped. The S. tyhimurium isolate was Multi-drug resistant (MDR) being sensitive to only Ciprofloxacin. Clinical signs observed when the MDR S. typhimurium was used to challenge 3-weeks old broiler-chicks were fever, watery-mucoid diarrhea, drop in feed consumption and reluntance to move with a mortality rate of 19 %. Post mortem examination of the S. typhimuruim infected broiler chicks revealed hepatomegaly with necrotic foci, enlarged spleen and enteritis. Recommended dose of Streptomycin (25mg/kg) could not cure the infection but when the drug was stabilized with Medicinal synthetic Aluminum magnesium silicate and was supported with Vitamin C (through feed) 75% of the recommended dose (18.75 mg/kg) cleared the resistant infection (100 % infection-reduction). The recommended dose even led to increase of the infection-load (-787.4% reduction) more than -656% infection-reduction of the untreated group, suggesting that Streptomycin treatment worsened the resistant infection. Survivors of the MDR S. typhimurium infection which were fed feed fortified with additional levels of Vitamin C attained a dressed weight of 1.5kg at 44days and higher cost of producing 1 kg of chicken (Ꞥ1,175.15) when compared with 1.5kg attained at 40 days at a cost of Ꞥ955.60/kg of chicken of the group of uninfected chicks fed with same feed with the additional Vitamin C supplementation..
TABLE OF CONTENTS
PAGE
TITLE PAGE ii
CERTIFICATION iii
DECLARATION iv
ACKNOWLEDGEMENT v
TABLE OF CONTENTS vi
LIST OF TABLES viii
LIST OF FIGURES ix
LIST OF PLATES xi
LIST OF ACRONYMS xii
ABSTRACT xiii
CHAPTER 1: INTRODUCTION 1
1.1 STATEMENT OF PROBLEM 2
1.2 SIGNIFICANCEOF STUDY 4
1.3 GENERAL OBJECTIVE 6
1.4 SPECIFIC OBJECTIVES 6
CHAPTER 2: LITERATURE REVIEW 7
2.1 CHICKEN EGG AND FOOD BORNE
ILLNESS 7
2.2 FOOD BORNE ZOONOTIC BACTERIAL
PATHOGENS 11
2.3 ANTIMICROBIAL RESISTANCE 31
2.4 ANTIBIOTIC RESISTANCE TO SOME SELECTED ORGANISM IN
POULTRY 40
2.6 STRATEGIES
TO LIMIT OF OVERCOME ANTIMICROBIAL RESISTANT STRAIN 41
CHAPTER THREE: GENERAL MATERIALS
AND METHODS 53
3.1 STUDY AREA 53
3.2
SAMPLE SIZE 53
3.3
STUDY ONE: SOCIO-ECONOMIC FACTORS AFFECTING USE OF ANTIBIOTIC IN POULTRY FARMS
IN ABIA STATE. 55
3.4
STUDY TWO: PREVALENCE OF BACTERIAL ISOLATE IN BROILER-CHICKS IN ABIA STATE. 56
3.5
INFECTION OF BROILER CHICKS INFECTED
WITH STREPTOMYCIN-RESISTANT SALMONELLA
TYPHIMURIUM AND TREATMENT WITH STREPTOMYCIN-MEDICINAL
SYNTHETIC ALUMINIUM MAGNESIUM SILICATE FORMULATION. 58
3.6 PROFITABILITY
OF RAISING BROILER CHICKS THAT SURVIVED SALMONELLA
TYPHIMURIUM INFECTION FOLLOWING TREATMENT WITH STREPTOMYCIN-MSAMS
FORMULATION. 60
CHAPTER
FOUR: RESULTS AND DISCUSSIONS 61
4.1
SOCIO-ECONOMIC FACTORS AFFECTING USE
OF ANTIBIOTICS IN POULTRY FARMS IN ABIA STATE. 61
4.2
PREVALENCE OF SALMONELLA SPECIES IN BROILER CHICKS FROM POULTRY FARMS IN ABIA
STATE. 68
4.3.
ANTIMICROBIAL SENSITIVITY PROFILE OF
FIELD SALMONELLA TYPHIMURIUM ISOLATES
TO ANTIMICROBIAL AGENTS USED IN POULTRY FARMS IN ABIA STATE. 68
4.4. EVALUATION
OF SALMONELLA SPECIE PROFILE OF THE
CHICKS BEFORE THE EXPERIMENT. 72
4.5.
PHYSICAL AND GROSS LESION OF SALMONELLA TYPHIMURIUM IN EXPERIMENTALLY
INFECTED BIRDS 72
4.6 RESPONSE
OF BROILER CHICKS INFECTED WITH STREPTOMYCIN-RESISTANT SALMONELLA TYPHIMURIUM AND TREATED WITH STREPTOMYCIN-MEDICINAL SYNTHETIC ALUMINIUM MAGNESIUM SILICATE
FORMULATION AND VITAMIN C. 72
47. COST
BENEFIT RAISING SURVIVOR BROILER-CHICKS TO 2KG LIVE WEIGHT FOLLOWING INFECTION
WITH RESISTANT SALMONELLA TYPHIMURIUM
AND WHEN TREATED WITH STREPTOMYCIN STABILIZED WITH MEDICINAL SYNTHETIC
ALUMINIUM MAGNESIUM SILICATE.
81
CHAPTER
5: CONCLUSION AND RECOMMENDATION 90
REFERENCE 91
APPENDIXES 123
LIST OF TABLES
4.1
Farm Characteristics and Management
of Poultry Farms in Abia State. 63 4.2 Socioeconomic
Factors affecting Use of Antibiotics in Poultry Farms in Abia State. 64
4.3 List
of Drugs Given to Birds in Poultry Farms Abia State 65
4.4 Farmers
Practice of Poultry Disease Prevention and Control in Poultry Farms Abia State. 66
4.5 Farmers
Knowledge of Antibiotics and Antibiotic Resistance in Poultry Farms in Abia State . 67
4.6 Prevalence
Salmonella Isolate from Apparently
Healthy Broilers in Poultry Farms in Abia State. 69
4.7 Mortality
Rate and Weekly Weight Gain of Broilers Infected with Streptomycin Resistance S. typhimurium and Treated with Graded
Doses of Streptomycin. MSAMS Formulation and Vitamin C. 76
4.8 Mean Colony Forming Units (CPU X 106/ml) of Streptomycin-Resistant Salmonella Typhimurium Per ml of Bile of
Chicks Treated with Streptomycin and Streptomycin-MSAMS
Formulation. 80
4.9 Infection
Reduction Rates (%) of Streptomycin-Resistant Salmonella Typhimurium in Broiler Chicks Treated with Streptomycin
and Streptomycin-MSAMS
Formulation. 82
4.10 Cost
Benefit Analysis 84
LIST OF FIGURES PAGE
2.1:
Mechanism of drug resistance by Gerard, D. W. 34
3.1 Map of Abia State. 54
4.1 Frequency of Antimicrobial
sensitivity of salmonella isolates
from Broiler birds in poultry farms in Abia State. 71
4..2 Mean daily body temperature of
broiler chicks experimentally infected with streptomycin resistance S. typhimurium and treated with graded
doses of streptomycin. MSAMS formulation and vitamin C. (P ≤ 0.05) 74
4.3
Mean daily feed intake of broiler chicks experimentally infected with
streptomycin resistance S. typhimurium
and treated with graded doses of streptomycin. MSAMS formulation and vitamin C.
(P ≤ 0.05) 75
4.4
Infection Reduction Rate (%) of resistant Salmonella
typhimurium Infection Treated with Streptomycin-MSAMS Formulation. 83
LIST OF PLATES
E
4.1:
Scanty colonies of salmonella species
on Salmonella Shigella Agar medium from 21 days old broiler chicks
prior to experimental infection. 70
4.2:
Insignificant growth of salmonella
isolates from apparently healthy broiler in poultry farms in Abia State on
Salmonella Shigella Agar. 73
4.3a:
Enlarged congested liver of broilers infected with streptomycin resistance S. typhimurium and not treated.
77
4.3b: Enlarged congested liver showing necrotic foci
of broilers infected with streptomycin resistance S. typhimurium and not treated. 78
4.4: Duodenum of broilers infected with streptomycin
resistance S. typhimurium and not
treated showing Haemorrahgic and catarrah enteritis. 79
.
LIST OF ACRONYMS
AMR: Antimicrobial resistance.
APEC: Avian pathogenic E. coli.
CAMP: Christie-Atkinson-Munch-Peterson.
CDC:
Center for disease and control.
DI: Day the birds were infected.
D1PI: Day 1 post infection.
D2PI: day 2 post infection.
D3PI: Day 3 post infection.
D4PI: Day4 post infection.
D5PI: Day 5 post infection.
D6PI: Day 6 post infection.
D7PI: Day 7 post infection.
DNA: Deoxy ribonucleic acid.
ESKAPE:Enterococcus faecium,
Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter
baumannii, Pseudomonas aeruginosa and Enterobacter spp.
CFU: Colony forming unit.
CLS: Clinical Laboratory Standards
Guideline.
CRE: Carbapenem-resistant
Enterobacteriaceae.
ELISA Enzyme-linked immunosorbent assay
ESBL: Extended-spectrum β-lactamases.
FAO: Food Agriculture Organization.
GBS: Guillain–Barre syndrome.
HA-MRSA:Hospital acquired Methicillin resistant Staphylococcus arueus
IU: Infected and untreated.
IS100: Infected and treated with
recommended dosage of Streptomycin (25mg/kg).
IS100M: Infected and treated with the
Streptomycin-MSAMS at the recommended dose (25mg/kg).
IS75MC: Infected and treated with
Streptomycin-MSAMS at 75% of Streptomycin (18.75mg/kg) and Vitamin C
(0.4mg/kg).
LMIC: Low- and medium-income countries.
LGA: Local government area.
NDR: Multdrug resistance.
MDR-TB: Multidrug-resistant Mycobacterium
tuberculosis.
MRSA: Methicillin resistant Staphylococcus arueus
MSAMS: Medicinal Synthetic Aluminum
Magnesium Silicate.
NCDC: Nigerian
center for disease control.
PALCAM:
Polymyxin acriflavin lithium-chloride Ceftazidime aesculin mannitol.
PCR: Polymerase chain reaction.
RT-PCR: Real-time polymerase chain
reaction.
UU: Uninfected untreated.
UUC: Uninfected broiler chicks on feed
sulemented with vitamin C(0.4mg/kg).
VRE:
vancomycin-resistant enterococci.
VRSA:
vancomycin-resistant S. aureus.
VISA:
vancomycin-intermediate S. aureus.
WHO: WorldHealth
Organization.
XDR-TB: extensively drug-resistant
Mycobacterium tuberculosis (XDR-TB)
CHAPTER 1
INTRODUCTION
Poultry is one of the most widespread
type of meat consumed worldwide (Heise et
al., 2015; Okorie-kanu et al., 2016; FAO, 2019). Poultry flocks
are often raised under intensive conditions using large amounts of
antimicrobials (AMs) to prevent and treat diseases as well as for growth
promotion (Agyare et al., 2018). An
antibiotic is a drug that kills (bactericidal) or stops the growth of bacteria
(bacteriostatic) (WHOa, 2020; Wikipedia, 2021). Antibiotics are chemicals
produced by microorganism such as bacteria and fungi (Jacob, 2015).
Antimicrobials are used in poultry farms for therapeutic, preventive or
prophylactic purposes and growth promotion (Agyare et al., 2018; Glasgow et al.,
2019; Wikipedia, 2021). In many developing countries, antibiotic-use in food
animals remains unregulated and there is evidence that lack of education on
proper use of antibiotics, limited awareness, in adequacies
in management and animal husbandry
practices, lack of hygiene and biosecurity measures have contributed to the
high use of antibiotics to prevent or treat disease outbreaks in poultry
(Oluwawemimo et al., 2016; Glasgow et al., 2019).
Antimicrobial
resistance (AMR) is a worldwide health concern and use of AMs in animal
production is a major cause of AMR in humans (Agyare et al., 2018). Antimicrobial resistant poultry pathogens may result
in treatment failure leading to economic losses as well as a source of
resistant bacteria phenotypes (including zoonotic bacteria) that may represent
a risk to human health (Van et al., 2012;
Okorie-kanu et al., 2016).
A number of studies have demonstrated increases in
resistance over time for bacterial organisms of Veterinary and public health
importance such as Salmonella species
(spp), Mycoplasma spp, Gallibacterium
spp, Escherichia coli spp, Ornitobacterium spp, Bordetella spp,
Enterococcus spp, Clostridium spp, Mycoplasma spp, Erysipelothrix spp,
Pasteuraella spp, Riemerella spp etc. (Anyanwu and Obette, 2015; Bortolaia et al., 2016). The indiscrimate use of
AMs in animal farming is likely to accelerate the development of AMR in
pathogens as well as in commensal organisms. In addition to the concerns due to
the emergence of AMR in bacteria from poultry production, there are also human
health concerns about the presence of AM-residues in meat (Reig and Toldra,
2008) and eggs (Goetting et al.,
2011; Okorie-kanu et al., 2016;
Agyare et al., 2018). Additionally,
AMR in poultry pathogens is likely to lead to economic losses resulting from
expenditure on ineffective AMs as well as the burden of resistant poultry
diseases.
1.1
STATEMENT
OF PROBLEM
A
lot of essential antibiotics are employed in poultry production in several
countries; threatening the safety of their products and increasing possibility of
development and spread of microbial resistance in poultry settings (Agyare et al., 2018).
A
surge in the development and spread of antibiotic resistance has become a major
cause for concern globally (Ventola, 2015; Oloso et al., 2018; WHOb, 2020). Over the past few decades, no major new
types of antibiotics have been produced and almost all known antibiotics are
increasingly losing their effectiveness against pathogenic microorganisms
(Agyare et al., 2018; WHOb, 2020).
It
is known, that worldwide, more than 60% of available antibiotics find use in
animal production for therapeutic and non-therapeutic purposes (Lander et al., 2012; Agyare et al., 2018). A lot of essential
antibiotics are employed during poultry production in several countries;
threatening the safety of such products (through antimicrobial residues) and
the increased possibility of development and spread of microbial resistance in
poultry settings (Lander et al.,
2012; Agyare et al., 2018).
Nigeria
is confronted with the burden of AMR (FMAEH, 2017; Oloso et al., 2018). The Nigerian center for disease control (NCDC) in
collaboration with other organizations is making efforts to develop an approach
to combat AMR using evidence based methods. Meanwhile, NCDC (2017) reported
that Nigeria has experienced huge resistance to antimicrobials in humans
especially in sepsis, respiratory and diarrheal infections.
Effectiveness
of currently available antibiotics is decreasing, due to the increasing number
of resistant strains of disease-causing infections (Ventola, 2015; Okorie-kanu et al., 2016; WHOb, 2020). In developed
countries, strict control of antibiotic use coupled with effective surveillance
of antibiotic resistance pattern in the population have successfully reduced
prevalence of AMR (Okorie-kanu et al.,
2016; WHOa, 2020). Again, the microbial loads of table eggs and other poultry
products are routinely evaluated before they are sold in developed countries
such as USA, Canada and Japan (Okorie-kanu et
al., 2016). The situation in developing countries like Nigeria is however
different. In Nigeria, antimicrobial agents are readily available to the public
in local drug stores without prescription (Okorie-kanu et al., 2016). Such practice has led to antimicrobial misuse
(Landoni and Albarallos, 2015), widespread AMR and treatment failure in
Veterinary and Medical practice. The problem is further compounded by the
virtual absence of effective legislation and programme for periodic evaluation
of the susceptibility of bacteria organisms of economic and public health
importance to available antimicrobials in Nigeria. Currently, the factors that
influence the choice and use of antibiotics by poultry farmers in Abia State is
poorly understood. To study the misuse of antibiotics by poultry farmers, it is
important to understand the factors that influence their attitude towards
antibiotic use (Xu et al., 2020).
1.2 SIGNIFICANCEOF STUDY
Poultry production is one of the
growing industries in Nigeria due to increased demand for animal protein
following rise in human population (Rashid et
al., 2003; Alahira, 2013). More than 180 million chickens are raised
intensively annually as a source of income, food or both (Alahira, 2013; FAO,
2019). Apart from direct consumption of eggs as food, eggs are also used in the
preparation of several commercial and homemade products such as mayonnaise,
cake, sandwich and pastries (Bettie, 2018). Poultry eggs are highly nutritive
but vulnerable to microbial contamination (Poliana et al., 2019; Rehault-Godbert et
al., 2019). The consumption of contaminated animal products give rise to as
food borne diseases (Ifedike et al., 2012;
Ovisogie et al., 2016).
Food
borne diseases cause an estimated 48 million illnesses and 3000 deaths in the
United States (Onyeneho and Hedberg, 2013; CDC, 2021). Reliable statistics on
food borne diseases are not available in most developing countries due to poor
or non-existent reporting systems (Ifedike et
al., 2012; Onyeneho and Hedberg, 2013). Thus, disease burden and
mortalities from food borne diseases could be higher in developing countries
like Nigeria where little or no control measures are put in place (Ifedike et al., 2012; Okorie-kanu et al., 2016). Normal intestinal commensals such as E. coli (Tadesse et al., 2012; Simoneit et
al., 2015; Luna-Galazet et al., 2016),
Enterococcus and Staphylococcus (Bortolacia et
al., 2015) as well as zoonotic pathogens such as Salmonella spp (Luna-Galazet
et al., 2016; Rehault-Godbert et al., 2019) are the most common and
leading causes of foodborne disease outbreaks worldwide (Ifedike et al., 2012; khedr et al., 2015; Okorie-kanu et
al., 2016; Rehault-Godbert et al., 2019).
They constitute major public health burden and represent a significant cost in
Medical and Veterinary care in many countries (Mclinder et al., 2014; Hoffmann et
al., 2015).
Antimicrobial
resistance is a global scourge (Micheal et
al., 2014; WHO, 2014). Antimicrobial resistance of pathogenic bacteria has
been partly attributed to the misuse of antimicrobials in Medicine and
Agriculture (Fasure et al., 2012;
Ventola, 2015; Xu et al., 2020; WHOc,
2020). Antibiotics are used by the poultry industry to enhance growth, feed
efficiency and to treat diseases (Agyare et
al., 2018; Wikipedia, 2021). Antimicrobial usage in poultry production
encourages efficient production allowing the consumer to purchase at a
reasonable cost, high quality meat and eggs (Donoggue, 2003; Agyare et al., 2018). However, the widespread
use of antimicrobial has made poultry a major reservoir of resistant microbial
(Crumpet al., 2002; Sule and Ilori,
2017; Agyare et al., 2018). The
reservoir of resistant bacterial in food animals implies a potential risk of
transfer of resistant bacteria from food animals to humans (Raufu et al., 2014; Xu et al., 2020). This transfer of resistant bacteria has severe
health implications including treatment failures which has led to some deaths
and increased cost of human and veterinary therapies (Marshal and Levy, 2011;
Agyare et al., 2018). Since
contaminated products of food animals are implicated as the most common cause
of foodborne infections, the microbial load of animal products should be
routinely evaluated before sales as done in developed countries (Oviasogie et al., 2010; Okorie-kanu et al., 2016). In addition, there is
need to evaluate possible mechanisms to enhance the effectiveness of existing
antimicrobials against human and veterinary resistant organisms. Such
mechanisms include antimicrobial stabilization leading to reduced rate of drug
metabolism, clearance and prolonged activity, in vivo. It had been shown that Medicinal Synthetic Aluminum
Magnesium Silicate (MSAMS) increased clearance percentage of Helignosomoides bakeri, Salmonella pullorum,
Plasmodium berghei (Ezeibe and Ogbonna, 2016; Ezeibe et al., 2019; Ezeibe et al., 2020).
1.3
GENERAL OBJECTIVE
The study is designed to determine
the factors that influence choice and application of antibiotics in poultry
farms in Abia State, evaluate the antimicrobial resistance profile of Salmonella typhimurium isolated from
broilers chicks from poultry farms in Abia State, and develop effective methods
of treatment of the bacterium to prevent development of AMR.
1.4 SPECIFIC OBJECTIVES
i. To determine socio-economic factors that
influence choice and application of antibiotics in poultry farms in Abia State.
ii.
To isolate and biochemically characterize Salmonella
isolate from broiler chicks in poultry farms in Abia state.
iii.
To evaluate the antimicrobial sensitivity of Salmonella typhimurium isolated from broiler chicks from farms in
Abia state, to commonly used antimicrobial drugs in poultry farms and identify
their resistant phenotypes.
iv.To evaluate the therapeutic effectiveness of
Streptomycin mostly being resisted when fortified with MSAMS and used to treat
chicks experimentally infected with resistant Salmonella typhimurium and fed withfeed supplemented with Vitamin
C/
v. Calculate the profitability of raising broiler
chicks thar survive Sallmonella paratyphoid following treatment with Streptomycin
mostly being resisted when fortified with MSAMS.
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