Emerging Infectious Diseases

 

Lecture 5 & Sample Questions

Y.J. Johnson

 

Antibiotic: Use, Misuse, and Bacterial Resistance

 

Objectives

1.      Discuss the roles of antibiotics in medicine and animal production

2.      Describe the mechanisms by which bacteria express and transfer antibiotic resistance

3.      Determine the role of antibiotic misuse in the development of resistant bacterial strains

4.      Discuss the implications of bacterial resistance for human and animal health

 

 

Antibiotic: Use, Misuse, and Bacterial Resistance

 

I.  How is this relevant to emerging infectious diseases ?  The term 'emerging disease' refers to:

A.     previously undiscovered pathogens, or

B.     known pathogens, previously controlled or treated that have:

1.     acquired new forms of virulence, or

2.     acquired new means surviving standard treatment

 

II.                   Terminology

A.     Antimicrobial - an agent that kills microorganisms or suppresses their growth

B.     Antibiotic - a chemical substance produced by a microorganism which has the capacity, in dilute solutions, to inhibit the growth of or kill other microorganisms

C.    Antibiotic Resistance - the ability of a microorganism to withstand the effects of an antibiotic that is lethal to most members of its species.  It refers to acquired genotypic changes which persist during cultivation in the absence of the antibiotic.

1.     It is a naturally occurring phenomenon

a)     antibiotic resistance genes have been characterized in coliforms from glacial water and ice estimated to be more than 2000 years old

b)     hence there is evidence of resistance prior to the use of antibiotics

2.     Antibiotic resistance is a  manifestation  of genetic diversity

3.     Those organisms with the genetic potential to withstand exposure to antibiotics that are lethal to most members of its species have a competitive advantage - natural selection.

4.     Antibiotic life-span

a)     Given the above, we can expect any antibiotic to have a limited effective lifespan

b)     The concern is that there has been a pattern of an increased rate of resistance developing within the past decade, thus antibiotics have a much shorter effective lifespan

 

III.                  Uses of Antibiotics

A.     In the US, antibiotic use is approximately evenly divided between human and agricultural use.

B.     The bulk of agricultural antibiotic use is for intensive livestock production

C.    Human and pet uses of antibiotics

1.     Therapeutic uses - treatment of an existing infection

2.     Prophylactic uses - preventing an infection

D.    Animal production uses of antibiotics

1.     Therapeutic uses - approximately 10% of the antibiotics used

2.     Prophylactic uses

3.     Growth promotion

a)     Approximately 90% of the antibiotics used in animal production are for prophylactic or growth promotion purposes

b)     Growth promotants consist of low-level, long-term mass administration of antibiotics usually via feed or water

c)      Growth promotants -

·        boost an animal's genetic potential for growth

·        result in improved feed conversion

·        reduce waste product output from intensive livestock production

·        there is a reported 3 - 5% increased rate of gain and feed efficiency attributed to the use of antibiotic growth promotants

E.     Crop Production - primarily spraying of fruit trees

 

IV.               Issues in the Use of Antibiotics

A.      How does exposure to antibiotics facilitate development of resistant strains?

1.      Given a population of bacteria exposed to an antibiotic; most of the bacteria are sensitive however a small number of spontaneously resistant mutants are in the population.  Those bacteria that are sensitive to the antibiotic die and the resistant strain survives and reproduces.  In the absence of the antibiotic the resistant strain continues to reproduce.  Over time the bacterial population is dominated by the antibiotic resistant mutant.

B.      Paradoxically both under and over use of antibiotics promotes development of resistance

1.    Examples of overuse of antibiotics in human and animal populations

a)     Treatment of mild self-limiting infections with antibiotics

b)     Use of powerful broad-spectrum antibiotics to treat an infection susceptible to a limited spectrum antibiotic.

c)      Long-term, low dose administration for prophylaxis (usually with debilitated, chronically ill patients)

d)     Prescribing antibiotics to patients with viral infections (often  in response to patient/client demands)

2.    How does overuse promote resistance?

a)     Exposing normal bacterial flora to antibiotics confers a selective advantage to resistant strains

b)     The resistant strains then colonize the host

c)      Resistant strains may now pass from the host into the environment

d)     In the environment, other bacterial species are exposed to the resistance genes and they are transferred to other species

3.    Examples of under-use of antibiotics in human and animal populations

a)     Failure to complete the full treatment regimen

·        Poor patient compliance - when the symptoms abate, the patient stops taking the medication

·        Economics/Access to medication - attempting to 'stretch' a single prescription to more than 1 illness or more than 1 patient

b)     Poor product quality - This is particularly important in developing countries

·        Expired products may be distributed or imported into countries with limited 'hard currency'.

·        Poor quality control - Substandard or adulterated products may be distributed or imported into countries without the infrastructure to adequately monitor and enforce product quality regulations.

c)      Group dosing - In intensive livestock production systems therapeutic group dosing of animals via feed or water results in uneven consumption of the medication, animals that consume less may not receive the desired dose.

4.    How does under-use promote resistance?

a)     As a result of under-use of a therapeutic dose of an antibiotic, there is a failure to achieve an adequate concentration of the antibiotic at the site of the infection.

b)     Alternatively, there may be a failure to maintain an adequate concentration of the antibiotic at the site of infection.

c)      As a result, only the most sensitive organisms are eliminated from the host.

d)     As in the over-use scenario, a selective advantage is then conferred to resistant mutants.

e)     The host is colonized by resistant strains

f)        Resistance genes may then be transferred to the hosts' normal bacterial flora and to the pathogen population.

g)     Resistant organisms may then pass from the host and spread to the environment.

5.    The use of antibiotics as growth promotants has become controversial due to concerns with antibiotic resistance.

a)     Growth promotants result in long term exposure to antibiotics during the animals' productive life.

b)     Growth promotants are group-dosed, usually in the form of feed additives

c)      Antibiotic growth promotants are used at sub-therapeutic levels.

6.    Effect of antibiotic growth promotants on host bacterial flora

a)     The effect is analogous to the under-use scenario: a sub-therapeutic concentration inhibits the sensitive flora; selects for the resistant flora; the resistant flora multiply within the host, pass into the environment and transfer resistance to other bacterial species in the host and environment.

b)     The impact of growth promotants is exacerbated by the fact that the host flora is exposed over a long span of time, perhaps through the entire rearing period.

 

V.                 How Do we Measure Resistance?

A.     In vitro - Tracking changes in the in vitro indicators of resistance allows one to monitor patterns of resistance and offers guidance in making clinical decisions.

1.     Minimum Inhibitory Concentration

·        The lowest concentration of an antibiotic that will inhibit visible growth of the isolate after incubation.

2.     Breakpoint Concentration

·        A specific concentration of antibiotic that is used to classify and isolate as sensitive or resistant.

3.     Zone of Inhibition

·        Using the disc diffusion method, the size of the zone containing no bacterial growth is used to determine the level of susceptibility to the antibiotic.

B.     In vivo - Clinical Outcome:  This is much more complex than in vitro determinations of resistance because it is dependent upon the interaction between Agent, Environment, and Host

1.     Agent factor: Type of bacterium determines whether the agent is intrinsically resistant to the antibiotic and its virulence factors.

2.     Environmental factors: Location of the infection in the host body; Distribution of the antibiotic in the host body; and Concentration of the antibiotic at the site of infection - all impact the efficacy of the antibiotic.  Infections in regions with low tissue perfusion or barriers to antibiotic diffusion may require increased dosages, more powerful antibiotics, or antibiotics with the diffusion characteristics necessary to cross physiological barriers.

3.     Host factor: Patient immune status: determines whether or not the infection can be successfully treated with a bacteriostatic treatment regimen versus a bacteriocidal treatment regimen.

 

VI.               Molecular Epidemiology of Resistance

A.     Intrinsic Resistance

1.      Species that are inherently resistant to specific antibiotics, i.e.

a)     This may be due to the permeability of the bacterium for a given class of antibiotic

b)     This bacterium may not possess the target site of action for a given class of antibiotic

2.      One clinical effect of intrinsic resistance is seen with Opportunistic Infections

a)     Use of an antibiotic may result in an overgrowth of inherently resistant organisms.

b)     Seen commonly when a patient develops Candidiasis (yeast infection/Thrush) secondary to antibiotic therapy.

B.     Acquired Resistance - When we speak of 'increasing antibiotic resistance' or resistant strains of bacteria as an 'emerging disease' we are referring to acquired resistance.

1.     Acquired resistance is when a bacterial strain that was sensitive to a particular antibiotic, develops resistance.  There are 2 ways in which this may occur:

a)     Mutation

·        This is a spontaneous event

·        It occurs regardless of antibiotic presence

·        It confers a selective advantage in the face of subsequent exposure to the antibiotic

b)     Acquisition of  New DNA - via 4 means: plasmid, transposon, bacteriophage, or transformation

 

VII.              Methods of Transfer of  Acquired Resistance

A.     Plasmids are self-replicating circular DNA that are smaller than the bacterial genome.  They can carry and transfer multiple resistance genes but they have a limited range of spread between bacterial species.

B.    Transposons are known as "jumping genes" and may be transferred from cell to cell.  They can either integrate into the recipient's plasmid or into the bacterial genome.  Unlike plasmids, they can spread resistance across a wide range of bacterial species.

C.    Bacteriophages are viruses that infect bacteria.  They are capable of transferring resistance as they infect the bacterial cell.  This method of transfer of resistance is important for Staphylococci spp.

D.    Transformation occurs when DNA that is released from a dying bacterial cell is taken-up by competent bacteria.  This method of transfer of resistance is important in the environment (i.e. soil).  It is an important factor in the transfer of penicillin resistance in S. pneumoniae

 

VIII.            Mechanisms of Acquired Resistance

A.     Modification or destruction of the antibiotic

1.     Beta-lactamases (over 200 types)

·        Enzymes that cleave beta-lactam rings

·        Confers resistance to penicillins, cephalosporins, carbapenems, monobactams

2.     Aminoglycoside-modifying enzymes (at least 20 enzymes)

·        Found in Gram negative and Gram positive orgranisms

3.     Others

·        Acetyltransferase -  chloramphenicol resistance

·        Esterase  -  macrolide resistance

B.    Reduced Uptake of the Antibiotic

1.     In Gram negative bacteria this is mediated via porins

·        Porins are aqueous channels that provide a hydrophilic route into the periplasmic space

·        Genetic mutations result in reduced number of porins or altered shape

·        As a result, permeability of the antibiotic into the bacterium is reduced.

C.    Increased Efflux of the Antibiotic

·        ATP binding proteins, membrane located efflux proteins and diffusion are 3 mechanism for increasing the efflux of antibiotic from the bacterial cell.

D.    Production of altered/new target site - examples

1.     Alteration in penicillin binding protein

2.     Production of a new cytoplasmic membrane protein

3.     Altered target enzymes

4.     Altered ribosomal RNA binding site

E.     Over-expression of the drug target - this is only described in Mycobacterium spp.

 

IX.               Current Public Health Concerns related to Antibiotic Resistance

A.     Enteric bacteria

1.     Multi-drug resistant S. typhimurium DT 104

·        Food borne or due to exposure to infected animals

·        90% of all S. typhimurium DT 104 isolated from humans are multi-drug resistant

·        resistant to chloramphenicol, streptomycin, sulfonamides, tetracycline

·        resistance to trimethoprim and fluoroquinolones are now being reported

2.     Campylobacter jejuni

·        Usually food borne disease

·        Poultry products are the most common source

·        Ciprofloxacin resistance has risen to 10%

·        The rises in resistance from human isolates has paralleled increasing resistance from chicken isolates

B.     Mycobacterium tuberculosis

1.     8 million new cases annually worldwide

2.     Tuberculosis is the most common infectious cause of adult death worldwide

·        3 million deaths annually

3.     Most deaths are NOT due to bacterial resistance but failure to properly treat susceptible strains.

4.     Multi-drug resistant Tuberculosis

·        Resistant to at least isoniazid and rifampicin

·        Now reported in 104 countries with a worldwide distribution

C.    Gram Positive Bacteria

1.     Methicillin resistant S. aureus

·        The infection is readily transmissible and 33% of the population carries the agent on healthy skin.

·        It is the second most common cause of bacteremia.

·        It causes superficial and deep skin, wound, and tissue infections.

·        An exponential rise in resistance has been reported between 1989 - 1996.

·        It is only susceptible to glycopeptides

2.     Strep. pneumoniae

·        It is the most common cause of community acquired pneumonia

·        It also causes meningitis and otitis media

·        It is the third most common cause of bacteremia

·        There has been a reported rising increase in in vitro resistance

3.     Enterococcus spp.

·        These organisms are considered normal gut flora

·        They are responsible for both community acquired and nosocomial infections

·        Vancomycin resistance has risen from 0.4% to 10% between 1989 - 1995

 

X.                 The Future

A.     World Health Organization  (WHO) Recommendations - In response to the growing concern over the increasing rate of acquired bacterial antibiotic resistance, the WHO held a meeting on the subject in Berlin, Germany in October of 1997.  The conference report, entitled "The  Medical Impact of Antimicrobial Use in Food Animals" makes the following recommendations:

1.     Terminate use of human antibiotics for growth promotion

2.     Establish threshold resistance levels

3.     Develop Risk Assessment and Post-Approval Monitoring Procedures

4.     Replace antibiotic growth promoters

5.     Record antibiotic import and export data

6.     Develop international residue standards

B.     Conclusions

1.     Increasing antibiotic resistance is a growing threat to public health

2.     Human and agricultural use and misuse of antibiotics has hastened the emergence of resistant bacterial strains

3.     Veterinarians and physicians must respond to the problem

 

 

Lecture 5 Sample Questions

 

1.      All of the following are TRUE statements EXCEPT:

 

a)     Treatment of self-limiting infections with antibiotics, is an example of antibiotic overuse.

b)     A broad-spectrum antibiotic should always be the first choice selected by a veterinarian or physician for treating an infectious disease.

c)      Exposure of normal bacterial flora to antibiotics selects for antibiotic resistance.

 

2.      Antibiotic growth promotants:

 

a)