Anti-infective drugs introduction.

Antibiotics their classification,

mode of action, and combination.
Microbial resistance and the risk of
antibiotic residues in food.
Infection is the invasion of a host organism's bodily tissues by disease-causing organisms,
their multiplication, and the reaction of host tissues to these organisms and the toxins they
produce. Infections are caused by microorganisms such as viruses, bacteria and fungi.
When infection attacks the body, anti-infective drugs can suppress the infection. Three types of
anti-infective drugs exist:
1. antibacterials (antibiotics and chemotherapeutic agents),
2. antivirals
3. antifungals
ANTIBACTERIAL drug is an agent that interferes with the growth and reproduction of bacteria.
An antibiotic is a chemical substance produced by a microorganism that inhibits the growth or
kills other microorganisms. Antimicrobial chemotherapeutic is a substance produced by
chemical synthesis that kills or inhibits the growth of microorganisms.
ANTIVIRAL drugs are a class of medication used specifically for treating viral infection. Like
antibiotics for bacteria, specific antivirals are used for specific viruses.
ANTIFUNGAL drug is a medication used to treat fungal infections such as ringworm
(dermatophytes), candidiasis (thrush), or serious systemic infections (cryptococcal meningitis).

ANTIBACTERIAL DRUGS (ANTIBIOTICS AND CHEMOTHERAPEUTICS)

The noun antibiotic was first used in 1942 by Dr. Selman A. Waksman, soil microbiologist.
Dr. Waksman and his colleagues discovered several actinomycetes derived antibiotics. The word
antibiotic will be used to describe: a chemical substance derivable from a microorganism that
kills or inhibits microorganisms and cures infections.
Each class and individual antibiotic acts in a different way and may be effective against either
a broad spectrum or a specific type of disease-causing agent.
Chemotherapy is the treatment of a disease by the use of pure chemicals (synthetic
antimicrobials) which have specific antagonistic effects on the organism causing the disease.
The action is achieved by interference with the metabolic processes of the organism (either
directly or indirectly), by antagonizing enzymes or preventing coenzyme formation, thus
starving the organism of a substance essential for normal life (sulphonamides).

SOURCES OF ANTIBACTERIAL AGENTS

A constant search is required to ensure that disease treatment can continue to be effective in the
face of antibacterial resistance. The sources of antibacterial agents are:
1. Mould and fungal metabolites (Penicillium spp.); bacteria (e.g. Streptomyces spp.,
Micromonospora spp., Bacillus spp.);
2. Semi synthetic variants of natural products (amoxicillin from benzyl penicillin).
3. Synthetic chemistry (sulfonamides, chinolones)

REQUIREMENTS FOR AN IDEAL ANTIBIOTIC AGENT

1.
2.
3.
4.
5.
6.
7.
8.
9.

Selective target target unique

Bactericidal kills
Narrow spectrum does not kill normal flora
High therapeutic index ratio of toxic level to therapeutic level
Few adverse reactions toxicity, allergy
Various routes of administration IV, IM, PO
Good absorption
Good distribution to site of infection
Emergence of resistance is slow

DIVISION OF ANTIBIOTICS
1. ACCORDING TO CHEMICAL STRUCTURE:
a. Beta-lactams (penicillins, cephalosporins, carbapenems and monobactams)
b. Tetracyclines
c. Amphenicols
d. Polypeptides and glycopeptides
e. Aminoglycosides
f. Macrolides
g. Lincosamides
h. Ansamycins (rifamycins)
i. Pleuromutilins
j. ATB with various structures (aminocoumarin and steroid).
2. ACCORDIG TO EFFECT ON MICROORGANISMS
a. Bacteriostatic antibiotic suppresses the growth of bacteria (tetracycline, tylosin)
b. Bactericidal antibiotic kills bacteria (penicillin, streptomycin, bacitracin)
3. ACCORDING TO MECHANISM OF THEIR ACTION
a. Cell envelope antibiotics cell wall synthesis inhibitors: -lactams, glycopeptides,
polymyxins
b. Protein synthesis inhibitors: tetracyclines, amphenicols, aminoglycosides, macrolides,
pleuromutilins, lincosamides, steroid substances (fusidic acid)
c. Nucleic acid synthesis inhibitors: rifamycins

4. ACCORDING TO THE SPECTRUM OF ACTIVITY

a. Broad-spectrum antibiotic one that is effective against a wide range of bacteria. It
also means that it acts against both G+ and G- bacteria (tetracyclines).
b. Slightly-broad spectrum (middle/mean broad) antibiotic (ampicillin, neomycin)
c. Narrow-spectrum antibiotic one, which is effective against only specific families of
bacteria (penicillin G, streptomycin).

ANTIBACTERIAL SPECTRUM
Antibiotics differ in the range of bacteria against which they are active. The most common
distinction is between those active against Gram-positive and Gram-negative microorganisms.
Typically a 'broad-spectrum' antibacterial will affect a range of micro-organisms, including
some Gram-positive and Gram-negative bacteria.
Thus benzylpenicillin affects only certain Gram-positive organisms (narrow spectrum),
mecillinam affects only certain Gram-negative organisms (narrow spectrum), while
oxytetracycline, amoxycillin and chloramphenicol affect a range of Gram-positive and Gramnegative organisms (broad spectrum).
The spectrum of certain antibacterials may also extend to include mycoplasmas (e.g. the
fluoroquinolones) or protozoa (e.g. dimetridazole).

ANTIBACTERIAL ACTION
Antibacterial action can be bacteriostatic or bactericidal.
The following antimicrobials at usual concentrations are generally bacteriostatic:
tetracyclines, amphenicols, macrolides, lincosamides, pleuromutilins, sulphonamides. These
produce stasis of bacterial growth in vitro. This means that, in vivo, the bacteria are made
susceptible to the body's defense mechanisms. Their successful use does, of course, depend on
these defenses being competent to achieve this effect, and this may be questionable in animals
that are very old, very young or greatly weakened and debilitated by the disease that is being
treated.
In general, the following common antimicrobials at usual concentrations are bactericidal:
penicillins (including semisynthetic), cephalosporins, aminoglycosides, polypeptides,
glycopeptides, potenciated sulfonamides, quinolones and nitrofurans. These produce actual
death of the cell in vitro, so when used clinically they should produce their therapeutic effect
without the aid of the body's defense mechanisms. Thus they are indicated in the
immunocompromised animals referred to above or where the immune system may be
depressed, e.g. in severe virus disease.
Classification of antimicrobials as bactericidal or bacteriostatic can also be misleading
because "bactericidal" drugs can be rendered bacteriostatic if sufficient drug concentrations are
not achieved at the site of infection.

MECHANISM OF ACTION OF ANTIBACTERIALS

CELL WALL SYNTHESIS INHIBITION
Some of the antibacterial compounds interfere with the cell wall synthesis by weakening the
peptidoglycan structures in bacterial cell wall, by this integrity of bacterial cell wall structure
weakens and eventually disrupts. Mammalian cells only have plasma membrane so these
antibiotics specifically target only bacterial cells. That is these antibiotics do not induce any
negative effect on the host mammalian cells. Several agents affect cell-wall synthesis, the most
important being penicillins and cephalosporins.
The major constituent of the cell wall of Gram-positive organisms is a mucopeptide and
that of a Gram-negative organism is a combination of a mucopeptide and a lipoprotein. The cell
wall is built up of layers of N-acetylmuramic acid and poly-N-acetylglucosamine molecules,
which unite in long strands with trailing polypeptides. In the final step cross linkages are
formed.
Gram-positive cell structure
The Gram-positive cell wall is thick and consists of 90% peptidoglycan. Teichoic acids link various
layers of peptidoglycan together. Teichoic acids also regulate the autolysin activity in this complex
equilibrium.
The cytoplasmic membrane (which defines the intracellular space) consists of a lipid bilayer;
intrinsic proteins which are hydrophobic (mostly enzymes involved in respiration and transmembrane
transport); extrinsic proteins which are hydrophilic; Penicillin-Binding Proteins (PBPs): periplasmic
space proteins involved in peptidoglycan synthesis (glycosyltransferase, transpeptidase and
carboxypeptidase activities)
Gram-negative cell structure
The outer membrane is made up of phospholipids, endotoxin or lipopolysaccharide (LPS) - plays
an important role in the antibiotic entry into the cell; proteins including the porins (complexes of three
proteins) form aqueous channels that provide a route across the outer membrane for all the water-soluble
compounds needed by the bacterium. The periplasmic space contains peptidoglycan 5-20% of cell
wall; various enzymes (in particular, -lactamases). The cytoplasmic membrane (which defines the
intracellular space) consists of: a lipid bilayer; intrinsic proteins which are hydrophobic (mostly
enzymes involved in respiration and transmembrane transport); extrinsic proteins which are
hydrophilic; Penicillin-Binding Proteins (PBPs) - periplasmic space proteins involved in peptidoglycan
synthesis (glycosyltransferase, transpeptidase and carboxypeptidase activities)

PROTEIN SYNTHESIS INHIBITION

Some of the antibiotic compounds inhibit bacterial cell multiplication by inhibiting protein
synthesis in them. Protein synthesis is a multi-step process. Majority of antibiotics inhibit the
process that occurs in the 30S or 50S subunit of 70S bacterial ribosome, this in turn inhibits the
protein biosynthesis.
Most of the antibiotics inhibits the formation of 30S initiation complex or altogether inhibits
the formation of 70S ribosome by the 30S and 50S ribosome subunits or they inhibit assembling
of amino acids into a polypeptide chain.
Tetracyclines block protein synthesis by preventing the binding of aminoacyl-tRNA in 30S ribosome
subunit. They interrupt the cycle by which amino acid is carried and attached to the ribosome. These
compounds block protein synthesis in both prokaryotic and eukaryotic system.
Aminoglycosides interfere with the formation of 30S initiation complex hence inhibits the protein
biosynthesis.
Amphenicols interrupt the transfer of a growing peptide chain to newly attached amino acids.
Macrolides interfere with the assembly of 50S subunit of ribosome hence inhibit the protein synthesis.

Lincosamides inhibit enzyme peptidyl transferase, hence prevent the protein synthesis. Macrolides and
lincosamides inhibit the movement of an amino acid to a donor site on the ribosome (translocation).

NUCLEIC ACID INHIBITION

Replication of the nucleic acids of the bacterial cell is prevented directly by nalidixic acid and
rifamycins and indirectly by the sulphonamides.
1. Effect on DNA. Sulphonamides ultimately deprive the cell of nucleic acid and the
presence of nalidixic acid prevents its replication.
2. Effect on RNA. Rifamycin specifically inhibits the bacterial enzyme concerned with the
replication of RNA. This is achieved by the binding of one molecule of the antibiotic to one
molecule of the enzyme.

CLINICAL USE OF ANTIMICROBIAL AGENTS - PRINCIPLES

Successful antimicrobial therapy is based on 4 principles:
1. Identification and characterization of the pathogen, including its antimicrobial
sensitivity, and selection of a drug based on the sites of infection and the lesions.
2. Effective concentrations of the indicated antimicrobial agent for a sufficient period at
the site of infection.
3. A dose rate, frequency, and route of administration of the antimicrobial agent, as
well as a duration of therapy, that maximizes the likelihood of a cure, prevents
relapse, and minimizes the risk of resistance without causing any harmful druginduced effects in the animal.
4. Specific and appropriate supportive therapy to enhance the animal's ability to
overcome the infection and associated disease conditions.
The immunocompetence of the animal should always be assessed, recognizing that
extraneous factors, stress, disease, malnutrition, and the effects of concurrently administered
drugs may influence the animal's ability to resist infection. Host defense systems are seriously
compromised in bone marrow depression, hypogammaglobulinemia, conditions in which
alveolar macrophages are incapacitated, tracheobronchitis, necrotic enteritis, starvation, and
other conditions. In addition, immunosuppressants (e.g., corticosteroids and antineoplastics)
compromise immune response. Any depression of immune capabilities can be expected to
modify the effectiveness of many antimicrobial agents, especially those with only bacteriostatic
activity.

REQUIREMENTS FOR SUCCESSFUL ANTIMICROBIAL THERAPY

CLINICAL DIAGNOSIS
Successful antimicrobial chemotherapy usually requires a specific diagnosis, even though a
reasonable preliminary diagnosis is often all that is possible, at least initially.
MICROBIOLOGICAL DIAGNOSIS
Treatment should be aimed at a specific pathogen whenever feasible. However,
polymicrobial infections are common. The ideal is a conclusive microbiologic diagnosis, but

frequently this must be presumptive (at least initially), and treatment must be based on
experience. Rational deduction may be necessary under field conditions. The examination of a
direct smear stained with Wright's or Gram's stain may help to establish what types of
pathogens are involved (gram-positive or gram-negative rods or cocci).
CULTURE AND SUSCEPTIBILITY TESTING
Isolation and characterization of the causative pathogen, susceptibility testing, and
determination of the MIC provide a sound foundation from which to select the antimicrobial
drug, as well as the dosage regimen. However, under field conditions, it is often difficult to
attain laboratory support for antimicrobial therapy.
MIC (Minimum Inhibitory Concentration) is the lowest concentration, which prevents
visible growth of bacteria; quantitative measure of the in vitro sensitivity of a particular
bacterium to a particular antibiotic.
APPROPRIATE SELECTION OF ANTIMICROBIAL AGENTS
Among the factors to be considered are the causative microorganisms, results of sensitivity
tests, pathogenicity of organisms, pathologic lesions, acuteness of infection, pharmacokinetics of
the drug indicated, expense, potential drug toxicity, organic dysfunctions (especially kidney and
liver function), and possible interactions with drugs administered concurrently.
Broad-spectrum antibiotics are properly used in the following medical situations:
1. Empirically prior to identifying the causative bacteria when there is a wide
differential and potentially serious illness would result in delay of treatment. This
occurs, for example, in meningitis, where the patient can become so ill that he/she
could die within hours if broad-spectrum antibiotics are not initiated.
2. For drug resistant bacteria that do not respond to other, more narrow-spectrum
antibiotics.
3. In super-infections where there are multiple types of bacteria causing illness, thus
warranting either a broad-spectrum antibiotic or combination antibiotic therapy.
CORRECT DOSAGE AND ROUTE OF ADMINISTRATION
The dosage selected should result in adequate therapeutic concentrations at the site of
infection for sufficient time without causing side effects or toxicity. For dose-dependent drugs,
higher dosages are more likely to enhance therapeutic success than shorter intervals. For lactam and other time-dependent drugs, therapeutic success appears to be greater if the
concentration remains above the MIC for about one-half to two-thirds of the dosage interval,
and efficacy is likely to be improved more by decreasing the interval than by increasing the
dose. The advocated dosage schedules should be carefully followed for at least 7 days (although
response should be apparent in 3 4 days for most infections), or longer if needed, to ensure
elimination of the pathogen and to prevent relapse, reinfection, or development of antimicrobial
resistance.
ANCILLARY TREATMENT, NUTRITIONAL SUPPORT AND NURSING CARE
Supportive treatment, optimal nutrition, and general nursing care are often critical for
successful management of infectious disease. Ancillary treatment might include the use of antiinflammatory agents, antidiarrheal preparations, expectorants, bronchodilators, inotropic
agents, urinary acidifiers and alkalinizers, immunopotentiators, and fluid and electrolyte
replacement. Attention should be given to caloric and nutrient intake, especially of protein and

vitamins. These nutrients play a cardinal role in immune responsiveness.

FACTORS INFLUENCING CLINICAL USE OF ANTIBIOTICS

A number of important details concerning absorption, circulation and metabolism of
antibiotics within the animal must be considered if the full benefit is to be obtained and the
antibiotics used correctly.
1. THE USE OF COMBINED ANTIBIOTICS AND JOINT USE OF ANTIBIOTIC AND OTHER
CHEMOTHERAPEUTICS.
2. THE BLOOD AND TISSUE LEVELS OF AVAILABLE ANTIBIOTICS ARE OF PARAMOUNT IMPORTANCE
WHEN ANTI INFECTIVE ACTIVITY IN VIVO IS BEING CONSIDERED.
The attainment of early high levels and their subsequent maintenance will depend on the
route of application and the nature of the formulation of the antibiotic. Thus the choice of
intravenous, intramuscular, subcutaneous, intraperitoneal or other parenteral route will
influence the rate of attainment and duration of activity. Further oral therapy may be used to
continue treatment or it may be used in some cases of less severity as the sole means of
treatment, particularly when an infection appears to be mainly in the gut.
3. REACTION WITH BLOOD AND TISSUE FLUIDS.
Some antibiotics such as penicillin are not decreased in potency by contact with blood,
serum, pus, cerebrospinal fluid, etc., whereas others such as streptomycin and sulphonamides
are influenced unfavorably by these fluids.
The absorption of some antibiotics from the intestine may be inhibited by the presence of
other chemical substances in the intestinal contents. Thus a high calcium level in the ingesta
(e.g. milk) will reduce the systemic availability of tetracyclines given orally. To overcome this
specially reduced calcium diet may be given for a short period.
4. PHYSIOLOGICAL BARRIERS.
There are a number of physiological barriers which some antibiotics will cross quite easily
but others only with difficulty. These barriers consist of:
a. The blood-brain barrier, which is a physiological division between the blood and
cerebrospinal fluid. Some antibiotics such as penicillin and streptomycin do not pass
the blood-brain barrier; in fact penicillin is actively pumped away from the
cerebrospinal fluid in a manner very similar to that in which it is eliminated in the
kidney. When the meninges are inflamed the pumping action is impaired and so
penicillins can enter the cerebrospinal fluid, although even then therapeutic levels may
not be achieved. Intrathecal injections should therefore be given at least to initiate
treatment. Other antibiotics such as oxytetracycline will also normally achieve
therapeutic concentrations in the cerebrospinal fluid only when the meninges are
inflamed. A further group, of which chloramphenicol is outstanding, passes the bloodbrain barrier fairly easily and achieves concentrations in the cerebrospinal fluid as
high as 50% of the blood levels.
b. The placental barrier does not normally resist the passage of antibiotics.
c. The intestinal barrier. When given orally many antibiotics are not absorbed into the

system in therapeutic concentrations. Streptomycin and neomycin are examples. The

reverse may also hold good. For example, when streptomycin is injected, very little is
excreted into the intestinal lumen except through the biliary system. This is important,
for in such cases only parenteral injection is suitable for treating systemic infections
and only the oral route can be used for treating intestinal infections. The failure of
neomycin to be absorbed is important in limiting its toxicity, which is quite
considerable by the parenteral route, but low by the oral route.
d. Serous membranes. Some antibiotics such as penicillin will not readily pass either
way across the normal pleural or peritoneal membrane, whereas others, for example
chlortetracycline hydrochloride, diffuse quite readily into the pleural and peritoneal
cavities.
e. Milk. The concentration of antibiotic in milk after parenteral administration depends
on the antibiotic involved. Benzylpenicillin and the semisynthetic penicillins are found
in the milk in considerably lower concentrations than in the serum, although the
concentration is raised in mastitis. Other antibiotics, such as spiramycin, tylosin and
the penethamate derivative of penicillin, are found in the milk in concentrations higher
than those in the plasma. Thus the latter are the most appropriate for systemic
treatment of mastitis. However, the intramammary route is generally preferred as it
gives direct access to the site of infection.
5. EFFECT ON INTESTINAL FLORA IN HERBIVORES.
Ruminants are highly dependent on the integrity of the rumenal microflora. Thus oral
administration of broad-spectrum antibiotics such as tetracyclines results in acute diarrhoea.
While this has been well known for some years, it has recently been recognized that
tetracyclines (by whatever route) can occasionally cause acute and even fatal intestinal
disturbance in horses. Although this is not yet fully understood, it probably results from a
change in the caecal flora, perhaps similar to that described below for hamsters and guinea-pigs.
Penicillin G, outstandingly safe in most species, has long been known to be toxic to guineapigs. This has been ascribed either to an allergy or to a disturbance in the intestinal flora. The
latter explanation now seems the most likely. It has been shown not only that penicillin is toxic
to the guinea-pig, but also that most broad-spectrum antibiotics produce the same effect, and
that similar toxicity is seen in the hamster.
It has been shown that the condition in hamsters is the result of overgrowth of Clostridium
difficile and that this organism is also the cause of the severe diarrhea in man. While it is not
clear if the same organism occurs in other species suffering from antibiotic-induced diarrhea,
this seems quite possible.
However, it is clear that extreme care should be taken in the use of antibiotics in herbivores.
In particular, tetracyclines, ampicillin, other penicillins, lincomycin and chloramphenicol are to
be avoided in hamsters and guinea-pigs. Ampicillin should also be avoided in rabbits, although
in this species tetracyclines seem relatively safe. Problems in herbivores are not avoided by
using the parenteral route, since toxicity can and does ensue in sensitive species.
6. ANTIBIOTICS AND IMMUNITY.
Some diseases do not recur because natural recovery from an attack results in the creation
and maintenance of adequate levels of immunoglobulins in the blood and tissues. If, however,
large doses of antibiotics or indeed of any other anti-infective agents are given, and the infection
is rapidly eliminated, the immunological reactions are not always properly stimulated and the

body is susceptible to further infection as soon as the antibiotic is withdrawn. Such situations
have arisen in the control of coccidosis by chemotherapeutic food medication.

COMBINATION OF ANTIBIOTICS
The administration of 2 or more agents may be beneficial in the following situations:
1. to treat mixed bacterial infections in which the organisms are not susceptible to a
common agent - to broaden the spectrum of activity,
2. to achieve synergistic antimicrobial activity against particularly resistant strains
(e.g., Pseudomonas aeruginosa),
3. to overcome bacterial tolerance,
4. to prevent the emergence of drug resistance,
5. to minimize toxicity, or
6. to prevent inactivation of an antibiotic by enzymes produced by other bacteria that
are present.
Additive or synergistic effects are seen when antibacterial agents are used in
combination, but antagonism may also emerge, sometimes with serious consequences.
Generally, bacteriostatic agents act in an additive fashion, whereas bactericidal agents are often
synergistic. However, the effects of several bactericidal antibiotics are substantially impaired by
simultaneous use of drugs that impair microbial growth or "bacteriostatic" drugs (eg, most
ribosomal inhibitors), so bacteriostatic and bactericidal combination may be antagonistic.
This antagonism results from the fact that many bactericidal antibiotics act only on dividing
cells. Thus bacteriostatic drugs, whose effect is to produce stasis of growth, will clearly interfere
with the bactericidal drugs. This is a general guideline only; many exceptions are known, and
confounding factors also play a role.
The most widely used combination of bactericidal drugs in Europe is penicillin G and streptomycin;
synergy can be demonstrated for this combination in vitro. A combination of bacteriostatic drugs which
produces synergy is that of sulphonamides with trimethoprim. This combination can even be bactericidal
under some conditions. The danger of combining bacteriostatic and bactericidal antibiotics was
dramatically demonstrated some years ago when it was shown that there was a higher mortality in
human patients with pneumococcal meningitis when they were treated with a combination of penicillin
and chlortetracycline rather than with penicillin alone.

Ideally, antimicrobial selection should be based on mechanisms of action that are different
and on spectra of activity that are complementary. -lactams are often selected because their
action is unique and not only complements other drugs but also facilitates movement of other
drugs through the damaged cell wall into the microbe. Examples of combination therapy for
mixed infections include the use of clindamycin, metronidazole, or the semisynthetic penicillins
for their anaerobic coverage in combination with aminoglycosides for their gram-negative
efficacy.
Preventing the development of resistance with combination antimicrobial therapy is best
exemplified by the use of carbenicillin or amikacin together with gentamicin or tobramycin for
the treatment of Pseudomonas infections.
Bacterial enzymatic inactivation of -lactam antibiotics, such as the penicillins and
cephalosporins, can be decreased by concurrent administration of a -lactamase inhibitor, such
as clavulanic acid or sulbactam.

RESISTANCE OF MICROORGANISMS TO ANTIBACTERIAL AGENTS

The emergence of bacteria resistant to antimicrobial agents within an animal population or
during therapy is of great concern. When resistance develops, a previously used therapeutic
approach may no longer be successful, and a suitable alternative antimicrobial drug must be
sought. Additionally, there frequently is concern from an epidemiologic and public health point
of view.
ANTIBIOTIC RESISTANCE
1. Natural (intrinsic) implies an intrinsic property in an organism that confers resistance;
found where an antibiotic is ineffective against a particular species of bacteria, such as
where benzylpenicillin is by nature ineffective against E. coli (this is a reflection of the
inability of benzylpenicillin to penetrate the bacterial cell wall, and so reach the target
enzymes situated there).
2. Acquired - suggests that an organism has obtained, by one mechanism or another, the
means to survive exposure to an antimicrobial agent; occurs when a previously
susceptible population becomes resistant following exposure to antibiotic. Most simply,
this occurs where small numbers of bacteria genetically able to resist the antibiotic were
contained within the original population. The removal of the susceptible bacteria by the
antibiotic gives the resistant bacteria a population advantage, they multiply and soon the
previously susceptible population becomes replaced by a resistant population. This socalled one-step process can be seen with streptomycin, where resistance development is
rapid.
MECHANISMS BY WHICH BACTERIA MANIFEST RESISTANCE
1. Organisms may produce enzymes, constitutive or inducible, which inactivate the drug
(e.g., penicillins, cephalosporins, aminoglycosides, chloramphenicol). Or defective
production of autolytic enzymes ("tolerance"), e.g., penicillins and cephalosporins. Or
decreased enzyme affinity, e.g., trimethoprim.
2. The permeability to or uptake of the drug by organisms may be decreased (inhibition or
changes in membrane transport systems to prevent entry of the antibacterial, e.g.,
aminoglycosides) or transport out of the cell may be increased (induction of membrane
transport systems to remove the antibacterial, e.g., tetracyclines).
3. Alteration of the drug receptor or binding site (specific configuration of target sites) may
result in reduced drug affinity at target loci (e.g., oxacillin, cloxacillin, macrolides,
lincomycin, streptomycin).
4. The organism may develop alternate metabolic or synthetic pathways to bypass or
repair the effects of the antimicrobial (e.g., sulfonamides, trimethoprim).
5. Development of impermeable cell walls with extremely narrow porins, e.g.,
Pseudomonas aeruginosa in response to many antibiotics.
In each of these cases, a modification of protein synthesis and enzyme activity is necessary to
confer resistance; thus, this adaptation is genetically determined.
Bacteria have 2 types of genetic structures that may confer resistance-chromosomes and
plasmids. Both consist of double-stranded DNA, and both are associated with the bacterial inner
cell membrane at some time. Plasmids are not essential for survival but do carry genetic
determinants that confer both antibiotic resistance and virulence on bacteria. Plasmid-mediated

resistance (R-factor or acquired resistance) is far more complex. Plasmids may contain 20-500
genes that can carry resistance to a number of different antibacterial agents (3-6 is common; up
to 9 have been recorded) and specific virulence factors. The 3 possible mechanisms by which
plasmids may migrate from one bacterium to another are transformation, transduction, and
conjugation.
MECHANISMS BY WHICH BACTERIA DEVELOP RESISTANCE
1. Mutation. Within a large population of bacteria, chromosomal mutations may occur,
which confer resistance either slowly, in a step-wise fashion with each succeeding
generation of the mutant more resistant or rapidly, in a single step in which the
bacterium is resistant after the initial mutation. Mutation is a random event.
Antimicrobials do not induce mutations but may exert a selecting out of resistant strains
by suppression of susceptible bacteria. Mutated bacteria are often metabolically
deranged and are at a selective growth disadvantage; they usually disappear with time
in the absence of the antimicrobial agent.
2. Conjugation. Certain Gram(-) bacteria undergo conjugation, a type of reproduction in
which genetic material is transferred from cell to cell via a pilus that is encoded by a
resistance transfer factor (RTF) on a plasmid (the DNA passes from the donor cell to the
recipient via a bridge formed during direct cell-to-cell contact). Resistance factors (R-factors) from plasmid DNA and/or chromosomal DNA may encode for resistance to
multiple drugs and may be rapidly transferred to the bacterial population. This is
termed infectious drug resistance or transferable drug resistance and has been
observed clinically in enteric infections with Salmonella spp., Shigella spp., and
Escherichia coli. General facets of conjugation make it an important process for gene
transfer under natural conditions. Many types of bacteria can act as recipients, and
resistance can pass freely from organisms normally saprophytic in the gut of animals to
pathogenic bacteria. Conjugation allows the passage of a number of distinct genes at one
time. Thus, resistance to several antibiotics, all mediated by different biochemical
means, may be acquired in a single step.
3. Transduction. The process of transference of drug resistant genes by bacteriophage
(that makes use of its specialized molecular equipment adapted for inserting DNA into
recipient bacteria) is termed transduction. Normally, it is phage DNA that is transferred;
in certain cases, however, some DNA from the episome in the bacterial cell replaces the
proper phage nucleic acid sequence. It may be important in the development of resistant
strains of Staphylococcus aureus.
4. Transformation. Bacteria may incorporate DNA encoding for drug resistance from their
environment after its secretion or release by resistant organisms (naked DNA seems to
pass from the donor to the recipient through the growth medium.). Acquisition of
resistance by this mechanism is relatively infrequent.
Genetic sequences capable of coding for resistance can migrate from a plasmid to a
chromosome and, then back to the plasmid. These sequences are then transpositional and are
known as transposons. A number of transposons responsible for the transfer of R-factor
resistance also have been isolated, characterized, and identified.
The clinical relevance of plasmid-mediated resistance principally concerns the following:

1.

intestinal infections, in which the reservoir of R-factors may be carried by saprophytic

flora in the gut;
2.
the use of low levels of antibiotics (as in animal feeds) or improper dosing regimens,
which may lead to a high incidence of R-factors in a given population (low subtherapeutic concentrations of antibiotics used as growth promoters in farm animals
may assist in the induction of resistance, too); and
3.
the indiscriminate use of antibiotics, which may eliminate the effectiveness of many
antimicrobial agents in the future
When antibiotics are removed from an environment containing a resistant population the
selection pressure is withdrawn and the previously susceptible bacteria (or at least those which
have survived) can now compete on equal terms with the resistant population. In some cases,
this may allow a gradual return to a susceptible population. Such reversion to susceptibility has
been demonstrated, for instance with respect to ampicillin resistance. On the other hand,
tetracycline resistance, once established, has been shown to persist even after the tetracycline
has been removed from the environment.
The use of antibiotics in food animals, including use as growth promotants, may contribute to
the transfer of resistance genes among bacteria and ultimately from food animals to humans,
where the organisms become pathogenic. The bacterial resistance created in the animal
following veterinary use of a drug or drug class may result in resistance to human drugs of the
same class. Whereas the organism developing resistance might be nonpathogenic, transfer of the
resistance gene to other bacteria in the human intestinal tract may result in a pathogenic
organism becoming resistant and ultimately in therapeutic failure in the human patient.
The following guidelines will help to minimize the emergence of bacterial resistance:
1. A broad-spectrum antibacterial agent should not be used if a narrow-spectrum agent is
also active against the causative organism.
2. Information regarding endemic infections and sensitivity patterns should be obtained
and considered when choosing an antibiotic.
3. Appropriate dose rates should always be followed.
4. When a combination regimen is used to prevent the development of resistant strains,
individual agents should be used at full dosage.
5. Antibacterials for topical application should be selected from those against which
development of resistance is uncommon.
6. To the extent that it is consistent with reasonable practice, every effort should be made to
use antibiotics only when the medical indications are clear and to avoid overuse of newer
agents when already available agents are effective.

RISK OF ANTIBIOTIC RESIDUES IN FOOD

The use of antibiotics in food-producing animals can lead to residue occurrence in food of animal
origin contributing to resistance or adverse effects/toxicity. Generally we recognize:
Therapeutic use of ATB - the risk of antibiotic resistance is relatively small where the
drugs are correctly used under veterinary supervision,
Prophylactic use healthy animals are being treated (antibiotics as feed additives) much resistance among bacteria banned in EU!
Metaphylaxis where disease is already present in a group of animals (e.g., diarrhoea in a

litter of pigs) an inevitable and reasonable use of the drugs.

ANTIMICROBIAL FEED ADDITIVES
Maintenance of healthy animals requires prevention of infection by pathogenic
organisms. In addition, specific alteration of a host's microflora may have beneficial effects on
animal production by alteration of ruminal flora, resulting in changes in the proportions of
volatile fatty acids produced during ruminal digestion. Thus, antimicrobial compounds may
improve production efficiency or growth of healthy animals fed optimal nutritional regimens.
Production-enhancing antimicrobial compounds can be classified as ionophore (e.g., monensin,
lasalocid) or nonionophore (e.g., virginiamycin, zinc bacitracin) antibiotics. Antimicrobial
compounds are administered in the feed at low dose rates relative to high doses required for
therapeutic effects.
There is circumstantial evidence that use of subtherapeutic doses of antimicrobials
creates selective pressure for the emergence of antimicrobial resistance, which may be
transmitted to the consumer from food or through contact with treated animals or animal
manure.
BAN ON ANTIBIOTICS AS GROWTH PROMOTERS IN ANIMAL FEED IN EU
Antibiotics have been widely used in animal production for decades worldwide. Added in low
doses to the feed of farm animals, they improve their growth performance. However, due to the
emergence of microbes resistant to antibiotics which are used to treat human and animal
infections ("anti-microbial resistance"), the Commission decided to phase out, and ultimately
ban, the marketing and use of antibiotics as growth promoters in feed. Antibiotics are now only
allowed to be added to animal feed for veterinary purposes. This decision was based on opinions
from the Scientific Steering Committee, which recommended the progressive phasing out of
antibiotics used for growth stimulation, while still preserving animal health.
Regulation (EC) No 1831/2003 of the European parliament and of the Council on additives
for use in animall nutrition:
Feed additives means substances, micro-organisms or preparations, other than feed material
and premixtures, which are intentionally added to feed or water in order to: favourably affect
the characteristics of feed, animal products, the colour of ornamental fish and birds; satisfy the
nutritional needs of animals; favourably affect the environmental consequences of animal
production, favourably affect animal production, performance or welfare, particularly by
affecting the GI flora or digestibility of feedingstuffs, or have coccidiostatic or histomonostatic
effect.
Antibiotics, other than coccidiostats or histomonstats, shall not be authorised as feed
additives.
Phasing out: Antibiotics, other than coccidiostats and histomonostats, could be marketed and
used as feed additives only until 31 December 2005; coccidiostats and histomonostats as feed
additives are phasing out of the use by 31 December 2012.
An EU-wide ban on the use of antibiotics as growth promoters in animal feed entered into
effect on January 1, 2006. The last 4 antibiotics which were permitted as feed additives to help
fatten livestock (monensin, salinomycin, avilamycin, flavofosfolipol) are no longer allowed to be
marketed or used from this date. The ban is the final step in the phasing out of antibiotics used
for non-medicinal purposes. It is part of the Commission's overall strategy and EU's food safety
strategy to tackle the emergence of bacteria and other microbes resistant to antibiotics, due to
their overexploitation or misuse.

PROBIOTICS
Probiotics promote the establishment and development of a desirable intestinal microbial
balance in the animal. There is a delicate balance between normal and pathogenic
microorganisms. This balance can be upset by poor husbandry conditions, disease, or stressors
(e.g. transport). Bacteria that produce lactic acid can, in general, be beneficial to the animal;
certain yeasts may also be beneficial.
Their ability to increase growth and promote health are claimed to be due to one or more of
the following factors: preventing colonization of the gut by pathogenic coliforms, altering GI
absorption rate, and inhibiting bacterial growth and influencing the balance of bacteria in the
gut.
The probiotic feed additives consist of selected strains of lactobacilli and streptococci that
alter the microbial species present in the GI system to the benefit of the treated animal.
Unicellular yeasts are also used. The production benefits are variable, and positive responses are
more likely when a stressful management change may result in a change in balance of gut
microflora.
Probiotics can help overcome the negative effects of certain conditions that detrimentally
modify the gut flora. Thus, they are useful in some cases to minimize GI upsets or to help
overcome stress due to weaning or transport. The unicellular yeast fungus may also have
beneficial effects on rumen fermentation and thereby improve digestion and feed efficiency.
The effect of probiotics in older animals may be reduced due to the well-established, balanced
population of micro flora that is less sensitive to minor detrimental husbandry challenges.