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Calming the Problem of Antibiotic Resistance
Mauj Yousif, Elise Maclean, Naz Pakkal, Kimberly Dias
11 March, 2014
Tutorial 1, TA- Rachel and Steve
Biology 1M03

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The Problem
It was only a few decades ago that antibiotics were known as wonder drugs that cured deadly
diseases. More recently, many antibiotics have become less effective and the amount of bacteria
resistant to antibiotics has increased significantly
Resistance can develop in four ways: mutation, inactivation, efflux and gene transfer. A
mutation in the DNA gyrase enzyme in bacteria prevents antibiotics from binding to its active site,
allowing the bacteria to continue to thrive. (Woodford N, 2007) When an antibiotic is used over a long
period of time, the frequency of bacteria who develop antibiotic resistance increases. For example,
Methicillin resistant Streptococcus aureus is a bacterium that is responsible for causing several difficult
to treat diseases in patients. To enumerate, MRSA is a current problem in hospitals as it has developed
immunity to certain antibiotics and can spread rapidly from one patient to another. According to a
study, 19,000 people died of MRSA during hospital stays in 2005 (Klevens et al, 2007).
This concept of antibiotic resistance originated in the 1930s. Sulfonamide drugs were the first
antibiotics that paved the path for the antibiotic revolution in medicine and started the mechanism of
resistance of bacteria. Sulfonamide resistance was originally reported in the late 1930s and similar
mechanisms of antibiotic resistance have appeared over the last 60 years. (Davies, 2010)
Over the years, different solutions have been attempted to reduce the increasing number of
resistant bacteria. One of the solutions that is still being used is referred to as a home-base strep test.
Patients often make the assumption that antibiotics are the cure for a sore throat although it is also
frequently caused by a virus which cannot be effectively treated with antibiotics. (Mersc, 2012) A
home base strep test is a diagnostic tool used to determine whether or not strep bacteria are present in
the patient’s throat. A positive result indicates the need for antibiotics. Although this test reduces the
unnecessary use of antibiotics, it is restricted in the sense that it has a single use as it only applies to
sore throat infections and is not a solution with long term benefits.
Another solution was offered by the FDA (Food and Drug Administration) that included

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guidelines for the use of antibiotics in the livestock industry. The excessive amount of antibiotics
administered to livestock creates a risk for antibiotic resistance to develop and affect the human
population. The guidelines proposed by the FDA were not enforced effectively and failed to calm the
problem of antibiotics.
Scientists agree that the failure of past attempted solutions have been ineffective due to the fact
that antibiotic resistance has more than one cause. This problem can still be reduced significantly by
controlling the way antibiotics are used. A few ways this can be done is by administering supplements
of alternative ways to consume antibiotics without extreme side effects.
The Solution
Over the past half-century, a large proportion antibiotics has been used toward livestock,
primarily for growth and therapeutic purposes. In 2001, it was estimated that 87% of all antibiotics
used annually in the USA were used for livestock, with only 13% in use for human therapeutic
purposes (Gilchrist, 2006). Australian import statistics also indicate that 55.8% of all antibiotics used in
the years 1992-1997 were used in stock feed, and used as growth promoters (Barton, 2000). This large
amount of antibiotics used for animals are used not only as growth promoters, but also to assist with
their health problems. For example, in Australia, tetracyclines, a class of antibiotics, are registered for
large-scale use to prevent respiratory diseases in pigs (Barton, 2000). To emphasize, livestock are also
fed antibiotics to assist in the digestion of certain foods, like corn, which disrupts their digestion.
(Wagner, n.d.). The intake of antibiotics results in serious threats and side-effects for livestock.
Antibiotic drugs destroy not only the harmful bacteria, but also the healthy bacteria present in the
intestines of the animal, which in turn makes them vulnerable to diseases. An important example of this
can be seen with the bacteria Clostridium difficile. It is a bacterium that causes intestinal conditions
such as inflammation of the colon and is the most frequent cause of the infectious diarrhea in hospitals.
It occurs mostly in patients who are taking certain antibiotics in high doses or over a long period of
time, which can destroy a person’s prebiotic bacteria found in the gut causing C. difficile bacteria to

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grow (Public Health Agency of Canada, 2013). Apart from this, the large amount of antibiotic drugs
present in livestock contributes significantly to the frequency of antibiotic resistance, as the presence of
drugs in animal tissue, as well as antibiotic residue, allow for the evolution of the resistant gene in
pathogens. (Wagner, n.d.). Similarly, in the evolution of MRSA, the mutant bacteria is resistant to
multiple antibiotics. There is therefore, an urgency to control the usage of antibiotics in livestock.
Controlling the amount of antibiotics that enter animal feed can yield substantial benefits. This
can be achieved by introducing a genetically modified version of cattle food, like corn, that contains
prebiotics. Prebiotics are specialized non-digestive foods that nourish healthy bacteria already present
in the intestine. One type of prebiotics are plant fibres: they are not digested, but allow good bacteria to
grow and thrive, as opposed to antibiotics, which kill off the healthy bacteria in the intestines and gut
(“What is a Prebiotic”, 2014). Administering animals, livestock and cattle in particular, with special
doses of prebiotics would reduce the required dosage of antibiotics in their diet by a large factor. The
reduction of antibiotics in animal feed would reduce the amount of bacteria that develop resistance to
these antibiotics, and can have beneficial implications on humans, the consumers. In particular,
removing antibiotics from animal feed will diminish the amount of harmful C. difficile bacteria in the
intestines, and can also eventually eliminate the lethal MRSA bacteria from circulation.
Research has shown that prebiotic use on preruminant and ruminant animals, such as cattle,
have had benefits on their health by enhancing the healthy bacteria in their rumen (Barton, 2000).
Prebiotics can be administered to cattle in several methods, depending on their need. In a study done by
Franklin et al in 2005, adult ruminants were given about 10 extra grams of prebiotics added to their
average daily intake, and the results indicated that the overall effect on them was positive (Barry, n.d.).
These results were similar to the results from another study which showed that the inclusion of
prebiotics in the milk of pre-ruminant calves led to increased body weight, and better feces consistency.
The increase in body weight may be due to increased fermentation at the small intestine followed by
increased flow of microbial nitrogen at the large intestine, as well as stable microflora composition at

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the rumen, small and large intestines of calves (Samanta et al., 2013). The results of these studies
indicate an overall positive result in livestock digestion due to prebiotic supplementation. A prebiotic
supplement, such as Prebiotin Prebiotic Fibre, can be introduced into animal feed such as corn, to
ensure proper digestion (“What is a Prebiotic...”, 2014). Farmers and cattle rearers should be highly
encouraged to administer the right amount of dosage of these prebiotic supplements to the cattle, in
order to minimize the dosage of antibiotics for animals. This strategy will have long term benefits, in
reducing the frequency of antibiotic resistance, but other strategies can show more immediate results.
Human Alpha-lactalbumin (HAMLET) is a breast milk protein that can selectively kill bacteria,
which acts on an individual basis to reduce antibiotic resistance. In some bacteria such as
S.pneumoniae and S.aureus it does this by binding to and stopping the activity of biological pumps
required for the flow of ions into and out of the cell. HAMLET is capable of stopping the two enzymes
that participate in glycolysis. Additionally, in the bacteria that HAMLET kills, it causes the same chain
of chemical reactions to occur as when bacterial cells undergo apoptosis. There is an inflow of calcium,
and the serine or threonine kinase is activated which results in the cell rupturing (“Protein Complex
Found ...”, n.d.). When HAMLET is used in combination with antibiotics, the concentration required
to treat infections is reduced significantly. In the absence of HAMLET the minimal inhibitory
concentration (MIC) of penicillin G was 0.01µg/ml for penicillin-sensitive S.pneumoniae, when using
HAMLET it was reduced five-folds to 0.002 µg/ml. Pneumococcal strains of bacteria which are
resistant to penicillin G require MIC of 4 µg/ml but in the presence of HAMLET the strain became
more susceptible to penicillin showing a 20-fold decrease in MIC to 0.2 µg/ml (Marks, Clementi and
Hakansson, 2012). Bacteria does not develop resistance to HAMLET. S.aureus developed resistance to
vancomycin, the antibiotics of last resort, although when used with HAMLET, S.aureus no longer
developed resistance. This is due to the fact that HAMLET is a naturally occurring protein complex
found in human breast milk and is not nearly as toxic as the high-powered antibiotics required to kill
already resistant bacteria. In fact, HAMLET causes bacteria to regain sensitivity to antibiotics (“Protein

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Complex ...”, n.d.). This suggests that mothers should be taught about the antibiotic properties of
HAMLET and be encouraged by healthcare professionals to breastfeed their infants. Along with the
use of prebiotics in animal feed, synthesizing HAMLET pills is another solution that can yield positive
results.
Another natural product that can be used in conjunction with HAMLET is Manuka honey
which inhibits wound infecting bacteria. In streptococci and pseudomonas, Manuka honey disrupts the
attachment of bacteria to tissues which is an essential step in the initiation of infections. This also
prevents the formation of biofilms which protect bacteria from antibiotics and allow them to become
resistant. When antibiotics are used with Manuka honey they can be made more effective against drug
resistant infections. Meticillin-resistant staphylococcus aureus (MRSA) can be made sensitive to
oxacillin when used combination with Manuka honey (“Honey Can Reverse.., n.d.). When Manuka
honey was used with rifampicin, rifampicin resistant MRSA did not develop (Mcdermott, 2013). A
Manuka honey supplement already exists, but it is clear that many individuals do not know about their
antibacterial benefits. Thus, individuals must be exposed to the honey supplement in the same way they
should be exposed to the HAMLET pills. The antibacterial component of Manuka honey is
methylglyoxal (MG), and the higher the concentration of MG present, the stronger the antibiotic effect.
The scale that rates the potency of Manuka honey is the Unique Manuka Factor (UMF). Honey with a
UMF rating of 10 or greater is considered to be effective as an antibiotic and is referred to as “Active
Manuka Honey” (“Manuka Honey: Medicinal”, n.d.). Therefore honey supplements with a UMF rating
of 10 or greater must be prescribed when individuals have bacterial infections.
Implementation and Cost
Before being able to implement the solution, the production expenses of the solution must be
examined. In this case, the cost of producing or synthesizing supplements with the HAMLET protein
should be considered as well as the importation of Manuka Honey capsules and prebiotic supplements
for the livestock.

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First, when evaluating the cost of the manuka honey it is found to be $40 for 500g on average
(Wedderspoon Organic, n.d). Pharmacies should have the capsules in stock so that when the doctors
prescribe it with antibiotics, antibiotic users will have a protection to resistant bacteria. When
assessing the cost of the prebiotic supplements needed, the average cost $29.99 for 80 capsules
(Prebiotin Prebiotic Bone Health, 2014). When importing both the prebiotic supplements and the
Manuka honey, the Canadian Government would pay the importation costs or taxes needed to import
the products into the country.
Secondly, in order to implement the use of the HAMLET protein, it is suggested that a
supplement be made out of the protein. Pharmacologists can produce HAMLET pills by isolating the
protein and purifying it. Filters and prepared gel filtration columns can be used to completely purify
this breast milk protein. This purified protein can then be replicated for large scale pill manufacturing,
and distributed to pharmacies (“Methods for Protein Purification”, n.d.). The journey to develop a new
drug is a long and hard process. There are several steps in this process which can be further divided
into seven phases. The first phase is preclinical research, where scientists will work to develop the drug
in question. After proven successes in living systems, a pharmaceutical company will file a report
called an Investigational New Drug Application with the FDA. Next, the FDA either approves the
drug or disapproves it and this step is what determines whether the drug will be able to be tested in
humans. If the drug is approved, it has to go through “Phase 1” drug trials, “Phase 2” drug trials and
“Phase 3” drug trials. Assuming that the drug is a success, a New Drug Application form is filed and
then if the FDA approves the drug, then it will be made available to patients (Fact Sheet New Drug
Development Process, n.d.). This entire process takes approximately twelve years to complete and on
average it can cost a company approximately $350 million (Herper, 2013). Normally, a pharmaceutical
company would cover the cost of creating a new drug.
In the case of HAMLET, once approved, doctors should be educated about the benefits the use
of HAMLET as an antibiotic and should prescribe it to their patients. It should be prescribed alone first

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as it can kill bacteria by itself. If this does not work to treat an infection, then antibiotics can be
prescribed in combination with the HAMLET supplement.
Ramifications
As no solution is ever perfect, there are many biological ramifications that must be considered
when striving to implement any solution. Future studies will have to be undertaken in order to measure
the effects of using HAMLET, Manuka honey and prebiotic supplements as suggested above.
Immediate results may be measured at the hospital level, where physicians and pharmacologists can
record the effects of their prescriptions on patients. For large-scale results, the effects of altering the
contents of cattle feed by increasing their prebiotic intake will have to be monitored over a long term
period. Overall, this solution to the problem of antibiotic resistance is one of long-term benefits,
although the effects of using HAMLET and Manuka honey can be seen sooner than the effects of
reducing antibiotic intake in livestock.
Some potential consequences of this solution is that synthesizing a new drug with the
HAMLET protein is found to be a long, expensive process and this may make it harder to execute the
solution. If HAMLET is approved for use, antibiotics will be used in lower dosages if at all. This will
decrease the costs that are necessary to produce treatments for bacterial infections. It is also impossible
to predict what repercussions might occur if the HAMLET supplement or Manuka honey is misused
once it is prescribed. Additionally, it is extremely challenging to monitor the misuse of the HAMLET
protein and the Manuka honey once it is prescribed as it is up to the person to use it correctly.
Although, if the public uses it for it’s intended purpose, there is clearly a positive consequence, the
issue of antibiotic resistance will be eliminated.
On the same note, reducing the antibiotics used on livestock and using prebiotic supplements
instead, result in positive ramifications. The use of prebiotics can help promote good digestion in
livestock and make them healthier. Notably, healthy animals are better equipped to resist pathogens
and as a result, animals require fewer antibiotics. Consequently, by diminishing the antibiotic

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resistance in livestock will bring about similar effects in the human population.
To elaborate, antibiotic resistant bacteria rely on the presence of antibiotics for survival. When
antibiotics are minimal or non-existent in the environment, the energy spent on antibiotic resistance
becomes a liability rather than a survival trait (Wagner, n.d.). Normal bacteria are able to live and
reproduce better than antibiotic-resistant bacteria, which makes it easier for widespread diseases to be
treated. Reduced antibiotics will not destroy as much healthy bacteria in the colon, and will therefore
suppress harmful bacteria like ,C.difficile which causes diarrhea (“Fact Sheet: Clostridium Difficile”,
2013). The reduction of antibiotics in animal feed is a solution with long term benefits, and will affect
the entire human population as a whole.
Conclusion
The problem of antibiotic resistance cannot be alleviated, but can be calmed. The frequency of
antibiotic resistance development in bacteria can be greatly reduced if the dosage of antibiotics
administered to livestock is decreased, as well as through the introduction of alternative methods for
reducing the need for antibiotics. Prebiotic supplements may be incorporated into animal feed to
improve their digestion, which will minimize their need for antibiotics. HAMLET and Manuka Honey
supplements may be used along with or instead of antibiotics, which also reduces the administered dose
of antibiotics. These supplements incorporated together can drastically reduce the frequency of
pathogens developing antibiotic resistance, simply because of a reduced amount of antibiotics exposed
to mutations.
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