Multidrug Resistant Staphylococcus Aureus in Poultry Droplets

Multidrug Resistant Staphylococcus Aureus in Poultry Droplets in Ebonyi State

Chapter One

Introduction

Staphylococcus aureus is a facultative anaerobic Gram positive coccal bacterium. It is non-motile, non-spore forming and catalase positive bacteria. The cell wall contains peptidoglycan and teichoic acid. The organism is resistant to temperatures as high as 50 0 c to high salt concentrations and drying. Colonies are large, 6-8 mm in diameter, smooth and translucent. S. aureus is frequently part of skin normal flora found in the nose and on skin and in this way 20 % of healthy human are carriers of Staphylococcus aureus(Van Belkum et al., 2009).

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Staphylococcus aureus is also a part of normal flora found in the axillae, the inguinal and perineal areas and the anterior nares. There are three patterns of carriage:

  • Those who always carry a strain
  • Those who carry the organism intermittently with changing strains
  • A minority of people who never carry Staphylococcus aureus.

Persistent carriage is more common in children than in adults. Nasal carriers may be divided into persistent carriers with high risk of infection & intermittent or non-carriers with low risk of infection (Van Belkum et al., 2009). The ability to clot plasma is the most widely used criterion for Staphylococcus aureus identification. Bound coagulase (clumping factor) reacts with fibrinogen to cause organisms to aggregate. The next factor is extracellular stahylocoagulase, which reacts with prothombin to form staphylothrombin that converts fibrinogen to fibrin.

1.1     Aims and Objectives

The aim of this study is to determine multidrug resistant Staphylococcus aureus in poultry droplets in Ebonyi state.

This study has the following objectives:

To isolate Staphylococcus aureus from poultry droplets, and to determine multi-drug resistant strain of Staphylococcus. aureus through susceptibility testing.

LITERATURE REVIEW

Staphylococcus aureus is a facultative anaerobic Gram positive coccal bacterium. One of the reasons for Staphylococcus aureus to be the most common staphylococci to cause staphylococcal infection is the presence of carotenoid pigment staphyloxanthin and this pigment acts as virulent factor, with an antioxidant action that helps the microbe evade death by reactive oxygen species used by the host immune system (Clauditz et al., 2006).

However, recent study carried by translational Genomics Research Institute revealed that half of the meat and poultry in US grocery store were contaminated with S. aureus with more than half of these bacteria resistant to antibiotic (Clauditz et al., 2006).

Staphylococcal infection is associated with the carriage of the organism. The organism is then disseminated through hand carriage to body (it is either through overt breaks in dermal surfaces, such as vascular catheterization or operative incisions, or through less evident breakdown in barrier function, such as eczema or shaving-associated microtrauma).

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Abscess is a distinctive feature of staphylococcal infection, which consists of a fibrin wall surrounded by inflamed tissues enclosing a central core of pus containing organism and leukocytes. From this point the organism may be disseminated hematogenously, even from the smallest abscess. The ability to elaborate proteolytic enzymes facilitates the process. This may result in pneumonia, bone and joint infection and infection of the heart valves.  

The organism produces toxins that can cause specific diseases or syndromes and can participate in the pathogenesis of staphylococcal infection. Enterotoxin- producing strains of Staphylococcus. aureus cause one of the most common food-borne diseases.

The organism causes staphylococcal scalded skin syndrome (SSS) in neonates and children, which is also known as Ritter disease. The organism produces an exfoliative toxin produced by strains belonging to phage group II. Initial features include fever, erythema and blisters which finally rupture and leave a red base.

The most feared manifestation of Staphylococcus. aureus toxin production is toxin shock syndrome (TSS). It was frequently associated with women using tampons during menstruation. Since the early 1990’s, half of the cases have not been associated with menstruation that produce the exotoxin TSST-1, but strains that produce enterotoxin Band C may cause 50 % case of non-menstrual TSS. These toxins are superantigens, T-cell mitogens that bind directly to invariant regions of histocompatibility complex class II molecules resulting in an expansion of clonal T-cells, next by a massive release of cytokines, which mediates the TSS.

However, it has been reported in hospitalized patients that S. aureus is intermediately resistant to vancomycin, which suggest that full resistance to vancomycin may eventually emerge. Although interspecies transfer of vancomycin resistance genes from vancomycin-resistant enterococcus was originally considered as the cause of this phenomenon, none of the species isolated have carried vanA, vanB, vanC1 vanC2 or vanC3 genes. Hence the clinical isolates with intermediate resistance to vancomycin were from patient who had undergone prolonged vancomycin therpy for methicillin resistant staphylococcus aureus (MRSA) (Garcia et al., 2011).

Recently, emergence of newly described mec genes and chromosomal cassettes from bovine sources has been described. Thus, the issue of antimicrobial resistance in Staphylococcus aureus continues to evolve (Garcia et al., 2011).

  • Virulence Factors of Staphylococcus aureus.

Staphylococcus aureus has the following virulence factors: Toxins, Protein A and Pigment.

2.1.1  Toxins

Staphylococcus aureus is capable of secreting several exotoxins, which are categorized into three groups. Many of these toxins are associated with specific diseases.

  • Superantigens

Staphylococcus  aureus has superantigen activities that induce toxic shock syndrome (TSS). This group includes the toxin TSST-1, which causes TSS associated with tampon use. This is characterized by fever, erythematous rash, hypotension, shock, multiple organ failure and skin desquamation. The staphylococcal enteroxins, which cause blood poisoning characterized by vomiting and diarrhea 1-6hrs after ingestion of the toxin, are included in this group (Lui et al., 2005).

  • Exfoliative Toxins (EFT)
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These toxins are implicated in the disease staphylococcal scalded-skin syndrome (SSS) which occurs most commonly in infants and young children. It also may occur as epidemics in hospital nurseries. The protease activity of the exfoliative toxin cause peeling of the skin observed with SSS.

  • Other toxins

Staphylococcal toxins that act on cell membranes are alpha toxin, beta toxins, delta toxin and several bicomponent toxins. The panto-valentine leukocidin (PVL) is associated with severe necrotizing pneumonia in children; the genes encoding the components of PVL are encoded on a bacteriophage found in CA- MRSA strains (Zhn et al., 2008).

2.1.2  Protein A

It is anchored to staphylococcal peptidoglycan pentaglycine bridges (chains of five glycine residues) by the transpeptidase sortase A protein A, an lgG binding protein, binds to the Fc region of an antibody.

In fact, studies involving mutation of genes coding for protein A resulted in a lowered virulence of Staphylococcus. aureus as measured by survival in blood, which has led to speculation that protein A contributed virulence requires binding of antibody Fc regions. Protein A in various recombinant forms has been used for decades to bind and purify a wide range of antibodies by immunoaffinity chromatography. Transpeptidase, such as the sortase responsible for anchoring factors like protein A to the staphylococcal peptidogycan are being studied in hopes of developing new antibiotics to target MRSA infection (Zhn et al., 2008).

2.1.3  Role of Pigment in Virulence

Some strains of S. aureus are capable of producing staphyloxanthin-cartenoid pigment that acts as a virulence factors. It possesses an antioxidant action that helps the microbe evade death by reactive oxygen species used by the host immune system (Lui et al., 2005).

Staphyloxanthin is responsible for its characteristics golden color. When comparing a normal strain of S. aureus with a strain modified to lack staphyloxanthin, the wild type pigmented strain was more likely to survive incubation with an oxidizing chemical, such as hydrogen peroxide, than the mutant strain was. Colonies of the two strains were also exposed to human neutrophils. The mutant colonies quickly succumbed, while many of the pigmented colonies survived. Wounds on mice were incubated with the two strains. The pigmented strains created lingering abscesses. Wounds with the unpigmented strains healed quickly. These tests suggest that the staphyloxanthin may be key to the ability of S. aureus to survive immune system attacks. Drugs designed to inhibit the bacterium’s production of the staphloxanthin may weaken it and renew its susceptibility to antibiotics (Lui et al., 2005). In fact, because of similarities in the pathways for biosynthesis of staphyloxanthin and human cholesterol lowering therapy was shown to block Staphylococcus. aureus pigmentation and disease progression in a mouse infection model (Lui et al., 2008).

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2.2.0  Staphylococcus aureus Resistance to Antibiotics

It was suggested  by Abigail and Dixie (2005) that penicillinase may have emerged as a defense mechanism for the bacteria in their habitats, as in the case of penicillinase-rich S. aureus living with penicillin-producing trichophyton. Search for a penicillinase ancestor has focused on the class of proteins that are capable of specific combination with penicillin. The resistance to cefoxitin and clindamycin in turn was speculatively attributed to Braine’s and Hartnell’s contact with microorganisms that produce them naturally or to random mutation in the chromosomes of Clostridium strains.

However, there is evidence that heavy metals and some pollutants may select for antibiotic-resistant bacteria, generating a constant source of them in small number (Abigail and Dixie, 2005).

2.2.1  Staphylococcal Resistance to Penicillin

This is mediated by penicillinase (a form of β-lactamase) production: an enzyme that cleaves the B-lactam ring of the penillin molecules, rendering the antibiotic ineffective. Penicillinase-resistant β-lactam antibiotics, such as methincillin nafcillin, oxacillin, cloxacillin, dicloxacillin and flucloxacillin are able to resist degradation by staphylococcal penicillinase.

2.2.2  Resistance to Methicillin 

Staphylococcal resistance to methicillin is mediated through the mec operon, part of the staphylococcal cassette chromosome mec (SCC mec). Resistance is conferred by the mecA gene, which codes for an altered penicillin-binding protein (PBP2a) that has a lower affinity for binding β-lacterms (penicillins, cephalosporins and carbepenems). This allows for resistance to all β-lactam antibiotics, and obviates their clinical use during MRSA infections. As such, the glycopeptides vancomycin is often deployed against MRSA.

Multidrug Resistant Staphylococcus Aureus in Poultry Droplets in Ebonyi State

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