Genetic Modification of Mosquitoes to Reduce Mosquito Borne Diseases

Research questions:
Which Method/s of Genetic Engineering Mosquitos Would be Best Suited to Reducing the
Cases of Mosquito Borne Diseases Across the Globe?

World studies theme: Health and Development

Subject lenses: Biology and Geography

Global issue: High Death Count Worldwide Caused by Mosquito Borne Diseases

Local manifestation: Implementation of each method based on cost and their effect on the
surrounding environment

Word count (without words excluded from word count): 3982

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Table of Contents
I-Introduction:...................................................................................................................3
II-Method of Collecting Data:............................................................................................5
III-Analysis.......................................................................................................................6
Method 1: Sterile insect technique (SIT)....................................................................................6
Method 2: Use of Genetically Modified Asaia Bacteria:............................................................9
Method 3: Use of Wolbachia Infection.....................................................................................13
VI-Discussion..................................................................................................................16
Discussing cost-effectiveness.....................................................................................................16
The Sterile Insect Technique........................................................................................................................17
Paratransgenesis Using Asaia Bacteria.........................................................................................................18
Wolbachia.....................................................................................................................................................18
Advantages and Disadvantages of Each Methods’ Costs.........................................................19
Discussing the Environmental Impact......................................................................................20
The Sterile Insect Technique........................................................................................................................20
Paratransgenesis Using Asaia Bacteria.........................................................................................................20
Wolbachia.....................................................................................................................................................21
Summary of Each Methods Impact on the Surrounding Ecosystem.......................................21
V-Conclusion...................................................................................................................22
VI-Evaluation..................................................................................................................23
Bibliography....................................................................................................................25

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I-Introduction:
Mosquito borne diseases pose a huge threat in our world today. These diseases include

malaria, zika virus, yellow fever, chikungunya, dengue, and more; most of which are spread

by mosquitoes of the Anopheles or Aedes genus, specifically Aedes. Aegypti (Centre for

Disease Control and Prevention, 2022). Unfortunately, current methods to prevent

transmission, such as bed nets, insecticides, and mosquito traps are not very effective and in

2018, approximately 830, 000 people died from mosquito borne diseases, mainly due to

malaria, zika virus and dengue. (WHO, 2022). Although there is an anti-malarial drug, which

kills the malaria parasite and prevents it from multiplying, it is becoming less effective as

prohibition proteins in the parasite prevent the drug from working (WHO, 2022).

Additionally, because mosquito borne diseases are generally found in less developed

countries, many people cannot afford these drugs or do not have access to them (Joy, 2018).

A study by Mara Ipa found that in Indonesia, over 20% of adults diagnosed with malaria, are

unable to receive anti-malarial drugs for treatment (Ipa, 2021). To tackle these diseases, more

complex methods involving genetic modification are needed, such as the sterile insect

technique (SIT), paratransgenesis using asaia, and the introduction of a Wolbachia infection.

SIT is a technique where many sterilized insects (by means of genetic modification) are

released into the wild to mate with the wild type mosquitoes without producing offspring

(Wang, 2021). Paratransgenesis is a method which prevents the transmission of malaria by

modifying bacteria from the mosquito to release proteins that inhibit the survival of

Plasmodium, before reintroducing them to the mosquito (Ratcliff, 2022). Lastly, Wolbachia

in infected mosquitos competes for resources with the disease-causing parasites, limiting the

mosquito from hosting them and thus reducing transmission. (WHO, 2022)

This essay will focus on each of the different methods and the research surrounding each one.

After the methods are analysed individually, they will be evaluated in terms of their cost and

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how much they damage the ecosystem. Finally, the methods will be compared and contrasted

to determine Which Method/s of Genetic Engineering Mosquitos Would be Best Suited to

Reducing the Cases of Mosquito Borne Diseases Across the Globe?

4
II-Method of Collecting Data:

1-The credited organizations below are used to obtain current and reliable published data:

 Research papers were from databases including Google Scholar, PubMed, Research Gate,

PNAS, and Science Direct.

 Information on the epidemiology of diseases was from the CDC database.

 WHO databases were used for information on health and development research.

2-This essay used secondary published data. Elements such as the price of each method, the

effects on the environment and the effects on the mosquitoes themselves were all considered

when looking for research.

3-The diseases focused on throughout this essay are malaria, zika, dengue fever, west Nile

fever and chikungunya as they cause the most deaths. Research about methods used to reduce

mosquito borne diseases were limited to those involving these diseases.

4-All data used for this essay was published in the last ten years.

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III-Analysis

Different uses of genetic modification to reduce mosquito borne diseases will be investigated to

determine its effectiveness and worldwide applicability and answer the research question. Finally, the

methods will be compared to conclude which one or combination of methods would be best at

reducing cases of mosquito borne diseases on a global scale.

Method 1: Sterile insect technique (SIT)

By releasing sterile male mosquitoes into the environment, the population of mosquitoes and the cases

of disease spread by them reduces (Wang, 2021). The SIT uses CRISPR/Cas-9 gene editing

technology, which permits scientists to remove the b2-tublin gene from male mosquitos, resulting in

infertile males.

This method has been used in the US, Malaysia, Brazil, and other parts of Latin America. It has also

been used for other insects and has proven to be effective (WHO, 2020). However, this is a temporary

solution as when the sterile mosquitoes are not released anymore, the population will eventually

rebound. This was demonstrated in a simulation where researchers used mathematical models based

on supplementary information from other sources and research projects to show the effect of SIT on

mosquito populations. (Stone, 2013)

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 Legend:

Red Line: Cage where trial

occurred

Blue Line: Second Cage where

trial occurred

Black Line: Mathematical model

of scenario

Grey Lines: Random Simulations

of Scenario
Populations of females, immatures and
WT males increase again

Release of sterile males stops so their

population decreases sharply
(Stone, 2013)

Figure 1: Graph Showing the Impact of the SIT on the Population size of mosquitos.

The solid green, black and red lines represent the populations of the wild type mosquitoes (male,

female, and larvae) and the blue line represents the population of the sterile males, which are released

weekly (explaining why their population fluctuates as time goes on). Whereas the dashed lines

represent the populations of the wild type mosquitoes if SIT had not been implemented. From the

graph, when the sterile mosquitoes are released the population of all three wild populations begin to

decrease steadily until the releases stop on day 240 of the simulation. Once this occurs, the

populations of wild mosquitoes begin to recover, eventually returning to normal. This simulation

shows clearly that populations of mosquitoes and their larvae will decrease as soon as the sterile male

mosquitoes are being released, then rebound when the release stops, increasing the spread of mosquito

borne diseases and making this method less suitable for reducing cases of these diseases across the

globe.

This technique was applied in real life in another study (Kyrou, 2018). In this study, researchers

modified A. gambiae mosquitoes to carry the dsxFCRISPR allele using CRISPR/Cas-9 technology (which

reduces the fertility of the mosquitoes). The modified mosquitoes were then crossbred to produce the

heterozygous offspring used for the trial. This trial had an initial modified allele frequency of 12.5%.

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The mosquitoes were then allowed to breed and lay eggs, of which 650 were randomly selected to

continue the next generation. In each generation, the larvae were analysed and screened for the

presence of the dsxFCRISPRh allele and its frequency. The data below shows the results of this trial.

(Kyrou,2018)

Figure 2: Dynamics of the spread of dsxFCRISPRh allele and effect on population reproductive capacity.

As seen in graph A, the frequency of the gene reached 100% in generation 7 (blue) and 11 (red).

These results match the model and most of the simulation predictions which also reach 100% in a

similar period (as seen by the grey and black lines).

However, in graph A, a dip in the red line is observed but not the blue line. Additionally, in both

graphs, the mosquitoes in the second cage reached the maximum (graph A) and minimum (graph B)

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of the graph before the mosquitoes in the red cage. However, the reasoning for both these occurrences

was not explained in the study. It can also be observed that as the gene frequency increased

throughout the generations, the egg output decreased, eventually reaching zero, causing the population

of the mosquitoes to decline as older mosquitoes die. The consequences of this are explained later in

this essay.

Although this method is used in some areas today, it is very carefully monitored to prevent a

population collapse, which isn’t very practical, reducing its suitability for worldwide application.

(WHO, 2022)

Method 2: Use of Genetically Modified Asaia Bacteria:

The process of paratransgenesis is explained using Figure 3 and the paragraph below it.

(1)

(3)

(4)
(2)

Figure 3: Diagram of the process of paratransgenesis in mosquitos.

(Ratcliff, 2022)

Paratransgenesis begins with the microorganism selection from the mosquito, which in this case is the

already existing Asaia bacteria (1). CRISPR/Cas-9 gene editing technology to cut out and replace a

section of genomic or plasmid DNA to create a transgenic bacterium (2). The bacteria are then

reinserted into the mosquito (3) where the modified gene is expressed to either release peptides that

interrupt the parasite transmission or dsRNA which silences the expression of the target gene from the

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parasite (4), preventing it from navigating the mosquitos’ body and accessing nutrients. In both cases,

the parasite development is blocked. The Asaia can be transmitted horizontally (through mating) and

vertically (through smearing of bacteria on eggs), which makes it appealing to spread the parasite

preventative throughout populations. (Ratcliff, 2022)

The ability for Asaia to spread across a population was tested in a semi-field study where mosquitoes

were kept in large cages (A, B, C), of 200 male and 200 female Anopheles Gambiae mosquitos, that

simulated their natural environments. Twenty paratransgenic mosquitoes were introduced into cage A,

thirty-five were introduced into cage B, and none were introduced to Cage C. The spread of the

bacteria was then monitored over the course of 20 days, showing the results below. (Mancini, 2016)

Figure 4: Graph Showing the horizontal transmission of the Asaiagfp across a mosquito population

(Mancini, 2016)

As shown in the graph, as the days go on, horizontal transmission increases, and so does the

percentage of mosquito midguts where the modified Asaia gfp is found. There is a quicker and higher

increase in Cage B where thirty-five modified mosquitoes were released and the maximum percentage

of guts with Asaiagfp is on average 81% (for males and females, after 20 days). However, the Asaia gfp

still managed to spread relatively quickly in cage A, reaching an average of 64% guts with Asaia gfp for

males and females, after 20 days. The control (Cage C) saw no spread as no modified bacteria were

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released there. The females in Cage A and Cage B always seem to have higher Asaia gfp percentages,

however the reasoning for this is not explained in the report. The increase of Asaia gfp as the days went

on shows that horizontal transmission is a good method to spread the bacteria across a population

(Mancini, 2016).

Another test was done, in the same conditions to check vertical transmission after twenty days,

showing the following results in Figure 5.

Figure 5: Graph showing the vertical transmission of Asaiagfp across a mosquito population

(Mancini, 2016)

Again, it is evident that the bacteria are vertically transmitted. However only about 40-50% of the

larvae inherited the bacteria from their parents who all hosted the bacteria. Of the newly emerged

larvae, 75% of the females born inherited the bacteria, whereas only about 45% of the males inherited

it. This isn’t as promising and could affect how this method is applied in an environment that isn’t a

cage. (Mancini, 2016)

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Another report which comments on an experiment indicates that this method works very well to kill

the Plasmodium parasite in mosquitoes (Nicholas J. Bongio, 2015). In the experiment the Asaia’s

siderophore receptor gene (siderophores are small compounds secreted by microorganisms) was fused

with scorpine, an anti-malarial peptide isolated from scorpion venom, or anti-Pbs21 scFv-Shiva1, an

immunotoxin, such that the Asaia would secrete these fusion proteins and their toxins would interact

with the membrane of the Plasmodium parasite, causing bacterial lysis. Both these strains of Asaia

were fed to mosquitoes which carried the malaria parasite Plasmodium berghei. After two weeks, the

mosquitoes were dissected and the number of oocysts (a cyst containing a zygote formed by the

Plasmodium parasite) in the mosquitoes’ midguts were counted. The results are shown in Figure 6.

Figure 6: Graph Showing the effect of different types of chemicals released by the Asaia on the number of
oocysts per midgut observed after 2 weeks. Black line in between dot clusters represents the median oocysts
per midgut for each chemical.
(Nicholas J. Bongio, 2015)

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As seen in the graph scorpine is most effective at reducing the oocysts and preventing the

Plasmodium from reproducing and thus being transmitted to humans as it has the lowest mean

number of oocysts per midgut. The P value is also very low, which supports the experiment’s validity.

Despite this, none of the toxins reach zero oocysts, meaning the plasmodium is still able to

survive/reproduce within the mosquito in both cases. This reduces its applicability as a method of

reducing mosquito borne diseases.

Method 3: Use of Wolbachia Infection

Wolbachia is an intracellular bacterium which can infect most arthropods. It can be passed down

when females lay eggs. When a mosquito is infected, Wolbachia competes with some types of

pathogens including those that cause malaria, dengue, zika virus and west Nile virus for fatty acids,

upregulate the mosquitos’ immune responses or interact with the pathogen itself to limit its ability to

infect, reproduce and spread. (WHO, 2021)

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Field trials by Peter A Ryan and colleagues in 2019 studied the impact of spreading a Wolbachia

infection in a population of Aedes Aegypti mosquitoes in northern Australia on Dengue cases. They

released 1,000 female mosquitoes infected with fluorescent Wolbachia in each of the 62 regions and

tracked the infection rate and dengue cases for 100 weeks.

Figure 7: Graph showing the impact of spreading a Wolbachia infection in a population of Aedes Aegypti mosquitoes in
northern Australia on % Ae. Aegypti infected (Graph A) and the number of areas with reported dengue cases during and
after release (Graph B). Grey boxes in graph A represent the interquartile range

(Ryan, 2019).

Graph A shows a sharp spike in the percentage of infected mosquitoes after the initial release of

Wolbachia. This is partly due to the continued release over the next 15 weeks, however even after the

releases stop, there is a steady increase until infection percentage reaches 100% at 60 weeks,

continuing to fluctuate until it steadies at 68 weeks. The interquartile range stays relatively small,

however there is a sharp increase at approximately 86 weeks. Graph B shows an immediate steady

decline in regions with dengue cases after the releases. This plateaus at 50 weeks, however, the cases

never reach zero, suggesting that this method is not 100% effective. (Ryan, 2019)

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A second semi field study done by several researchers uses Wolbachia infected mosquitoes. (Liu,

2022). The researchers transinfected 200 female Aedes. Aegypti mosquitos with the Wolbachia strain.

They were also infected with four different strains of dengue, (DENV 1-4) and left to incubate for 7

and 14 days. After, they were dissected to check how much the dengue was able to replicate. The

results are summarised in the Figure 9, showing the plaque forming unit (PFU/ml) to represent the

number of viral particles capable of forming plaque within one millilitre. The two values (e.g., 2.9 x

104 and 177) for each region of the body (Midgut, Saliva Glands, Saliva) and strain of dengue

represent the PFU/ml for the Wildtype controls (upper value) and the Wolbachia mosquitoes (lower

value). (Liu W. L, 2022).

Figure 8: Table Showing the impact of wALB Wolbachia on the number of viral particles capable of forming
plaque in 1ml (PFU/ml), 7 and 14 days after infection. PBM: Post Blood Meal (7PBM = 7 days Post Blood
Meal)

(Liu, 2022).

The wAlB Wolbachia strain was very effective in reducing the replication of the dengue as the

PFU/ml for all body areas decreased significantly from the wild type mosquitoes to the modified

mosquitoes. This is evident as the difference in the upper and lower values of PFU/ml for each body

area and strain of dengue is very high. (e.g., from 1.9 x 10 4 PFU’s/ml in the saliva glands of wild

mosquitos after 7 days, versus the 69 PFU’s/ml in the saliva glands of wAlbB-Tw mosquitos in the

same amount of time). This difference was most significant in the Midguts and Saliva Glands which

had much higher PFU’s/ml in the wild type mosquitoes than the saliva. The data also shows how

depending on the dengue strain, the results of Wolbachia intervention can vary, as DENV-1 and

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DENV-2 are most affected by the Wolbachia and have lower PFU’s/ml than DENV-3 and DENV-4,

which may be more resilient strains. However even though the Wolbachia reduces the replication of

dengue significantly, it is not 100% effective as the dengue is still present, meaning the dengue can

still be transmitted and infect more people. This reduces its appeal for application in the real world.

(Liu W. L, 2022)

VI-Discussion

Discussing cost-effectiveness

Mosquito borne diseases are most commonly found in the regions of Sub-Saharan Africa, Latin

America, The Caribbean, South/Southeast Asia, and Australia. (Centre for Disease Control and

Prevention, 2020). Many countries in these areas are not very economically developed, and have

medium to low GDP per capita, as explained using Figure 10.

GDP per capita demonstrates a country’s prosperity and wealth, thus it can be used to determine

approximately how readily the country can invest in these methods for the prevention of mosquito

borne diseases.

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Figure 10: Graph Showing the approximate GDP per capita values for each country in 2020

(Our World in Data, 2020)

By looking at the map, it is evident that most of the regions/countries mentioned above, where

mosquito borne diseases are most prevalent, have a medium/low GDP per capita (represented by the

light colours in Figure 10). Thus, the methods used should be cost effective.

The Sterile Insect Technique

A cost effectiveness assessment done by multiple organisations including the World Health

Organisation and the International Atomic Energy Association compares SIT to previous methods

used to reduce mosquito-borne diseases. This calculation looks at costs of a combined SIT and

incompatible insect technique (IIT) which is like the sterile insect technique but uses Wolbachia to

render mosquitoes infertile rather than genetic modification. The HC males named by their

developers, Hydrocarbon Search Partners, were released periodically and the cost of their release was

calculated per hectare (ha) of space they are released in.

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The cost assessment is also based on how the cost of this method is predicted to change over the next

few years.

Figure 11: Cost Assessment of Sterile Insect Technique done by WHO

(World Health Organization, 2021)

Paratransgenesis Using Asaia Bacteria

Unfortunately, because this method is so new, there is little relevant data on the economic effect this

method would have on the countries it is applied in.

Wolbachia

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 A report by PLOS Global Public Health looked at the economic impact of dengue in Singapore from

2010 to 2020 and how this value would change if Wolbachia was used to decrease transmission of

disease. The results are shown in Figure 12.

Figure 12: Table showing the estimated economic cost of dengue and the costs that could be averted at different levels of Wolbachia
efficacy

(Soh, 2021)

Advantages and Disadvantages of Each Methods’ Costs

Method of Reducing Cost Advantages Cost Disadvantages

Mosquito Borne Illness

Sterile Insect Technique Trials are inexpensive, with an average total Method is not sustainable so long-term

cost of 13.55 USD per hectare per year costs will accumulate

Costs of this method are predicted to

decrease even further in the next 10 years

(from 216.5 USD /ha/year in the first two

years, to 115.5 USD /ha/year in the 8 years

after)

Paratransgenesis Using N/A N/A

Asaia Bacteria

Wolbachia At minimum efficacy (40%), this method Even at maximum efficacy (90%), over

still saves 328 million USD over the last ten 250 million USD is spent due to dengue

years. This value increases as efficacy over the last 10 years, which is a lot.

increases.

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 The method is sustainable, so costs don’t

recur as frequently.

Figure 13: Table Showing the Advantages and Disadvantages of Each Methods' Costs

Discussing the Environmental Impact

The Sterile Insect Technique

The use of SIT to control mosquito populations could be effective in reducing the spread of mosquito-

borne diseases, however, it could also have negative impacts on the environment. Mosquitoes and

their larvae serve as a food source for many other species and their decline could disrupt the food

chain. Additionally, mosquitoes are important pollinators and a decrease in their population could

affect the health of plant species.

Paratransgenesis Using Asaia Bacteria

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A study found that in most species of bacteria in the Asaia genus, there is a gene called the pyrethroid

hydrolase gene that plays a crucial role in establishing a symbiotic relationship between the bacteria

and mosquitoes. (Commandatore, 2021). However, when paratransgenesis occurs, the DNA sequence

that releases proteins that interrupt the hosting of Plasmodium parasites also promotes the pyrethroid

hydrolase gene, leading to resistance to pyrethrin’s, which is a key ingredient in insecticides. This

causes a problem as around 38% of the world's land is used for agriculture and it is distributed mainly

in South and East Asia, Australia, and tropical areas of Latin and South America. (Food and

Agriculture Association of the United Nations, 2020). This is shown in Figure 14 below.

Figure 14: Map showing the distribution of agricultural land use in 2018. (Ritchie, 2020)

Wolbachia

According to research, it appears that Wolbachia has minimal impact on mosquitoes or their

ecosystem. The National Environment Agency has stated that Wolbachia is safe for humans, wildlife,

and plant life, as it is a naturally occurring bacterium found in over 60% of insect species and cannot

survive outside of an insect host. Additionally, humans and animals have had long-term exposure to

Wolbachia without any reported illnesses. (Natural Environment Agency, 2022)

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 Summary of Each Methods Impact on the Surrounding Ecosystem

Method of Reducing Impact on Surrounding Ecosystem

Mosquito Borne Illness

Sterile Insect Technique Decreased mosquito populations take away a food source from their predators, causing

their populations to decline as they die of starvation. This could start a ripple effect

which would spread across the ecosystems in which SIT is applied.

Mosquitos are pollinators so as their populations decrease, so do the number of plants

fertilised which could lead to declining plant species/populations.

Paratransgenesis Using Paratransgenesis contributes to insecticide resistance in mosquitos so they can infiltrate

Asaia Bacteria farms and spread diseases between plants, harm livestock (consistent blood-sucking

leads to loss of weight and health issues) and harm humans who run the farms. This

could lead to decreased food yield (contributing to food shortages) and poverty for

farmers. (Commandatore, 2021)

Wolbachia No known effects on surrounding animal or plant life. (Natural Environment Agency,

2022)

Figure 15: Table Showing the Impact of each Method on the Surrounding Ecosystem

V-Conclusion

The essay concludes by identifying the best method for reducing mosquito-borne diseases from the

research gathered. Each method has its own advantages and disadvantages. The SIT (Sterile Insect

Technique) and Wolbachia technique are well-researched and have been implemented in several

countries, while Paratransgenesis, which uses new technology, has not been trialled as much.

Therefore, from a reliability perspective, the SIT and Wolbachia method are the most effective.

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However, a disadvantage of the SIT method is that it is not sustainable as the population of

mosquitoes rebound as soon as the release of sterile mosquitoes is halted. On the other hand, both

Wolbachia and Paratransgenesis using Asaia use bacteria that can be transmitted between offspring

and spread in populations, making their effects longer lasting. This was demonstrated in an Australian

study in which the effects of Wolbachia continued up to 84 weeks after the release of the infection

stopped. However, another study found that traces of Wolbachia were still found after the infection

had been implemented, indicating that the mosquito could still spread the dengue, reducing the

sustainability of the method. Likewise, when Asaia was modified, it was found that although it greatly

reduced Plasmodium development, the mosquito could still carry and spread the disease. Additionally,

studies have shown that only 40-50% of offspring inherit the Asaia bacteria from their parents,

indicating that the method of using Asaia is more sustainable than SIT but less sustainable than using

Wolbachia. In terms of cost, both the SIT and Wolbachia methods show promise, but it is difficult to

compare the cost data as the report by WHO on SIT looks at the direct cost of implementing the

method, while the data for Wolbachia looks at how much cost could be avoided if the method is

applied. Despite this, the SIT method is less sustainable, which would result in higher long-term costs

than Wolbachia. There was no data found for Paratransgenesis, likely due to its recent development.

Furthermore, the ecological impact of the SIT and Paratransgenesis using Asaia is quite severe,

whereas Wolbachia has no known impacts. The SIT reduces mosquito populations, affecting the food

chain, and plants pollinated by mosquitoes, leading to a decrease in animal and plant populations. On

the other hand, Asaia, when modified expresses a gene which increases insecticide resistance in

mosquitoes, allowing them to spread into farmland, harming wildlife, humans, and potentially causing

detrimental damage to the economy. Due to its limited testing, a solution to this issue has not been

found yet, whereas for the SIT it is possible to still use it while closely monitoring the surrounding

environment.

Overall, while all these methods are promising, the Wolbachia method stands out the most. Although

it is not as trialled or cost effective as the SIT, it is the most sustainable and poses the least risk to the

surrounding ecosystem. It is also trialled enough to be approved and applied. However, in less

economically developed countries, the SIT could be used as a short-term solution while economic

23
growth continues, as it has low short-term costs. On the other hand, Paratransgenesis was eliminated

as it isn’t trialled nearly enough and still needs to forgo a lot of development before it can be safely

used and is not as sustainable as the Wolbachia method.

VI-Evaluation

It is important to evaluate the significance of sources and data when drawing conclusions for this

essay. It is assumed that these are the only methods that need to be considered to reduce mosquito-

borne diseases around the world. However, there may be other methods that were not considered in

this essay, which discredits the conclusion about the most suitable method.

Secondly, although there is data to support each argument, most of it is derived from cage trials and

simulations rather than real-world data. This reduces the validity of the data as although the cages

simulate the outside world as closely as possible by controlling many variables such as humidity, food

availability, etc. there are always factors that cannot be accounted for. Even though a certain method

may be successful in a caged trial, it may not fare so well outside a lab environment. This could affect

the conclusion.

As mentioned previously, all data was published within the last 10 years which increases the validity

of this essay.

This report also focused primarily on Biology and Geography, yet this question also involves subjects

like business, economics, politics and more, which should be studied further. Although the

conclusions drawn from this essay can support decisions made regarding mosquito borne diseases,

much more research needs to be done.

24
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