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Thursday, 7 March 2019

ISOLATION, IDENTIFICATION AND CHARACTERIZATION OF Escherichia coli and Staphylococcus aureus AS SOURCES OF SKIN INFECTION.




ABSTRACT
Escherichia coli and Staphylococcus aureus are serious causes of a variety of community and hospital-acquired infections. E coli is one of the most common nosocomial pathogens that cause urinary tract infection (U.T. Is ) and enter colitis. S. aureus is  also  an  etiological  infection  agent  responsible for significant levels of morbidity and mortality. Escherichia.  coli accounts for 17.3% of infection behind Staphylococcus aureus (18.8) in recent years, the emergency of resistant Staphylococcus.  aureus and resistant Escherichia.  coli strains to many antibiotics has been observed worldwide. These have become a major concern in global public health invigorating the need for new antimicrobial compounds. Their taxonomies, morphology and biochemical characteristics. Habitat and growth characteristics, the caused infections, their treatment and resistances to antibiotics.
Furthermore, Thirty people out of which ten Students from the Oke Ogun Polytechnic Saki and the remaining twenty five are from Appinite junction area, Appinite , best impact area, Appinite,  Longest villa area, Ajegunle bike stand and Ajegunle, Gbawojo Market inside the Saki town were selected randomly for sample collection, sterile swab sticks were used to take the samples from the selected individuals. Results of the test yielded significant growth of E coli and S aureus
Isolation and identification of microorganisms particularly Escherichia coli and Staphyloccocus aureus were carried out using streak plate method, biochemical and identification test and also biostatistician analysis were also done on result of the isolate.
Key Words:  E. coli, S. aureus, Antibiotics resistance, Infections, Taxonomy, Isolation,
Number of words: 250.


TABLE OF CONTENT
TITTLE                                                                                                      i
CERTIFICATION                                                                                                ii
DEDICATION                                                                                           iii
ACKNOWLEDGEMENT                                                                          iv
TABLE OF CONTENT                                                                              v
CHAPTER ONE
1.0 Introduction                                                                                          1
1.1 Sources of skin infection                                                                      1
1.2 Location of Skin Normal Flora                                                             5
1.3 Life on the surface of the skin                                                               6
1.3.1 A Bacterial Sneeze, the Nose                                                             7
1.3.2 A Mouthwash Away                                                                         7
1.4    Normal Skin Flora                                                                                       8
1.5   Aims and Objective                                                                          9
1.5.1 Statement of Problem                                                                        10
1.5.2 Justification                                                                                       10
1.5.3 Scope of the study                                                                                      11
CHAPTER TWO
2.0 Literature Review                                                                                 12
2.1 Overview of skin flora on (E. coli)                                                                 14
      Taxonomy
      Morphology and Biochemical Characteristics
        Habitat and Growth Characteristics
         Serological characterization of Escherichia coli
2.1.2  Pathogenic of Escherichia coli                                                           17
2.1.3  Neonatal meningitis caused by E. coli                                                        17
2.1.4  Escherichia coli and Antibiotic Resistance                                        17
2.1.5  β-lactic antibiotics                                                                                      17
2.2.0  Sulfonamides                                                                                    18
2.2.1  Trimethoprim                                                                                   19
2.2.2  Quinolones                                                                                        19
2.3    Overview of skin flora on (S. aureus)                                                         20
2.3.1  Staphylococcus aureus                                                                    20
Brief history                                                                                              
          Taxonomy
          Morphology and growth characteristics
          Natural habitat
2.3.2  Virulence factors of Staphylococcus aureus                                               23
2.3.3  Pathogenesis of S. aureus                                                                           24
Deep-seated infections                               
Bacteraemia
Metastatic infections
Toxin-mediated diseases
2.4 Antibiotic resistance in Staphylococcus aureus                                             26
2.4.1  Pathogenic of Staphylococcus aureus.                                                        27


CHAPTER THREE
3.0     Materials and Methods                                                                    29
3.1     Materials to be used                                                                          29
3.2     Research Methodology                                                                     30
3.2.1 Collection of Sample                                                                         30
3.2.2  Culture Media                                                                                   31
3.2.3  Sterilization of materials                                                                             31
Glassware
Inoculating chamber
3.2.4- Media and Diluents                                                                           31
3.2.5  Culture of organisms                                                                        32
          Preparation of media
3.3.1  Cultivation of bacteria                                                                      32
3.3.3  Streak Plate Method                                                                         33
Procedure for Streaking Method
3.3.4           The isolates were characterized using the following test                             33
3.3.5  Cultural and morphology characteristics                                         34     
3.4     Characterization and Identification of Microorganism                      34
3.4.1  Staining procedure                                                                                     34
3.4.2    Simple staining                                                                               34
3.4.3  Gram Staining                                                                                   35
3.4.4  Acid Fast Staining                                                                                      35
3.5     Biochemical Test                                                                               35
3.5.1 Catalase Test                                                                                    35
3.5.2  Oxidase Test                                                                                    36
3.5.3   Indole Test                                                                                      36
3.5.4   Methyl Red And Voges—Proskauer Test                                       36
3.5.5    Citrate Test                                                                                     37     
3.5.6    Statistical Analysis                                                                         37
           CHAPTER FOUR
4.0       Result and Discussion                                                                     38
4.1        Result                                                                                             38
4.1.1      Occurrence of Isolates                                                                                      39
4.1.2      Table I: Characterization and Identification of bacteria isolate from    normal skin flora within Saki town                                                                39
4.1.3      Table II:  Microbiological counts of E. coli and S. aureus within Saki   metropolis (Using ANOVA method).                                                               40
4.1.4       Table III: Occurrence of   bacteria species in some of normal skin
flora of human within Saki town                                                      41
4.1.5      Table IV: Characterization and Identification of E. coli and
          S. aureus                                                                                           42


CHAPTER FIVE
5.0     Discussion                                                                                        43
5.1     Findings                                                                                            45
5.2      Recommendation and Conclusion                                                   47
5.2.1  Recommendation                                                                              47
5.3     Conclusion                                                                                        48
REFERENCES
         











                  

CHAPTER ONE
1.0            INTRODUCTION
Sources of skin infection
Infectious diseases are leading cause of global mobility and mortality (Ryan, 2014). In 1990, infections cause 16 million deaths, and in 2010, the number of deaths had fallen to 15 million [W.H.O 2013]. The spread of infectious diseases result as much from changes in human behavior. Including lifestyle and land use pattern, increased trade and travel, and inappropriate use of antibiotic drugs as form mutations in pathogens [Ngwai, 2010].Staphylococcus aureus and Escherichia coli are a major cause of various humans and animals infections. The first causes skin and soft tissues infections, surgical site infections and bone and joint infections. Staphylococcus aureus is a common cause of hospital acquired bacteraemia and it is associated with hospital acquired respiratory tract infections [Ryan, 2014]. E. coli is the most common cause of urinary tract infections (UTI) in humans [Foxman, 2010], and is a leading cause of enteric infections and systemic infection [Ngwai, 2010].
The systemic infections include bacteriamia, nosocomial pneumonia, cholecystitic, cholangitiz, and peritonitis, cellulites, osteomycelitis and infection arthritis. E. coli is also leading because of new meningitis [Kim, 2011]. A wide range of antimicrobial agents effectively inhibit the growth of E. coli. The β-lactase, fluorguninolones, amino glycosides and trimethoprim-sulfame thoxoizole are often used to treat community and hospital infections due to E. coli (Fast M-Bunzeluk, 2012). But antimicrobial resistant isolates, especially those that are fluoroguinolone resistant and those producing bit ended spectrum β-lactamonses have increased significantly during the 2,000s and in certain area in many nococomial and community-acquired E. coli are now resistant the several important antimicrobial classes (Ortega, 2009).
            Penicillinase-resistant penicillin’s, flucloxacilin, dicloxacillin) remain the antibiotics of choice for the management of serious mehticillin susceptible S. aureus (M55A) infections, but first generation cephalosporin (Cefaxiporin, Cephalothin and cephalexin), clindamycin, lincomycin and erythromycin have important the suspected roles in less serious MSSA infections such as skin and soft tissue infections or in patients with penicillin hypersensitivity. All serious MR5A infections should be treated with parental vancomycin or if the patient is vancomycin allergic, (Wang H. Dzink, 2011).
            Antibiotic’s resistant staphylococci are major public health concern since the bacteria can be easily circulated in the environment. Infections due to methicililn-resistant Staphylococcus aureus (MR5A) have increased worldwide during the past twenty years (Pitout, 2012). Some report of S. aureus isolate with intermediate or complete resistance to vancomycin portend a chemotherapeutic era in which effective bactericidal antibiotics against this organism may no longer be readily available (Pitout, 2012). Multiple during-resistant S. aureus have been frequently recovered from foodstuffs (Acc. Ferreira, 2011), water and bio-film formulation (Doughti, 2011). Nasal mucosa of humans  (Karezmarezyk, 2012).
            This paper aims to review the taxonomies, morphology and biochemical characteristics, habitant and growth characteristics, the caused infections the treatment and resistance to antibiotics of these two bacteria.
            Even though an individual has a “normal” skin flora it often happen that during his or her life there are fluctuation in the composition of this florin due to general health conditions, diet, hormonal activity, age and many other factors.
            The micro organisms most o which are bacteria comprise the normal skin flora also termed the normal micro biota. They inhibit mainly the inner surfaces of the body such as the mucous membrane of the oral cavity and genitourinary tract most are highly adopted to survival and growth in this area despite physical and chemical condition that discourage many other kind of micro-organisms.
            Therefore the termed normal skin flora implies that these microbes are harmless, and for the most part that they do not cause disease and are oven beneficial. Most are commensally, they benefit from the association with the host they benefit the host in some fashion while thriving in the host body. It is of interest to learn what these beneficial effects are and how they can be lost due to change in the normal skin flora cause by the use of the antibiotics or other means,.
            In addition some normal skin flora organism can be opportunistic pathogen that is they may cause infection if tissue occurred or at specific site or if the resistance of the body to infection is decrease. This is especially importance because in recent years there has been a rising incidence of infection and from this micro organism arises.


FREQUENT VISITOR
THE MAJOR MICROBIAL SYMBIOTANT FOUND ON OR IN THE SKIN OF HUMAN.
COURTESY OF B.C BLOCK AND J DUCAS, MAN MICROBES AND MATTER (2010).




1.2     Location of Skin Normal Flora
Microbes that colonize the human body during birth or shortly thereafter remaining throughout life are referred to as normal floral (Scheupelin, 2010). Normal floral can be found in many sites of the human body including the skin (especially the moist areas, such as the growing and between the toes), respiratory tract (particularly the nose).  Urinary tract and the digestive tract (primarily the mouth and the colon). On the other hand, areas of the body such as the brain, the circulatory system and the lungs are intended to remain sterile (microbe free).
Figure 1: Location of normal microbial floral. Each of these areas of the body contain their own microenvironment and various inhabitants of microbes. The human body provides many unique environments for different bacterial communities to live. In this context, scientists refer to the human body as the host. A positive host’s microbe relationship is usually described as either mutuality or communalistic. In mutualism both the host and the microbes benefit.
Which is in contract to commensalisms, where one partner of the relationship Bonnets (usually the microbe) and the other partner neither bereted nor harmed. In many cases it may be difficult to establish whether a particular host microbes relationship should be considered mutuality or communalistic, since scientists are only beginning to understand the role of normal flora in human health. In other words, individual microbes may be carrying out important functions within our bodies that we have not yet discovered. Just as host—microbe relationships can be positive or neutral, they can also be negative. Such a host—microbe relationship is usually described as parasitic or pathogenic. In a parasitic relationship the microbe bonnets at the expense of the host and similarly in a pathogenic relationship the microbe causes damage to the host. In both cases the cost to the host can vary from slight to fatal. Whether a host—microbe relationship is “positive” or “negative” depends on many factors. And in most cases the relationship will actually remain positive. The host provides a niche and nutrition for the colonizing microbe and the microbe, occupies a space that a potential parasite or pathogen might otherwise colonize. In these cases microbial communities may even aid in digestion or synthesize nutrients for the host. However, life is not always perfect, and in certain situations good-standing members of your normal flora can cause disease or invading pathogens can displace them. The result will be disease. To illustrate some of these scenarios let’s take a closer look at microbial communities found in different areas of the human body.






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