Earth and Life Science

REPORT ON TECHNIQUES AND PRODUCTS OF GENETIC ENGINEERING

INTRODUCTION

Pigs are one of the most common animals that we know. They have made many contributions to people’s lives,
especially the farmers. However, it is shown that pigs also have a negative effect on us -- humans and marine life.
According to studies, pigs are one of the main contributors to water pollution. Moreover, scientists were able to solve the
problem in preventing the pigs from causing bigger problems. But is it really okay?

According to the Environmental Protection Agency or EPA, Nitrogen, and phosphorus are two nutrients that
pollute the water in more than 100,000 miles of streams and rivers in the United States, as well as nearly 2.5 million acres
of lakes, reservoirs, and ponds. According to them, Swine manure is one of the main factors responsible for this
large-scale contamination. Swine manure is a valuable by-product of the livestock and poultry industry that can be used
as a fertilizer. It is a product of the pig farming industry and is rich in both nitrogen and phosphorus. Pigs are omnivorous
mammals with thick skin that is usually sparsely coated with short bristles. They have a stout body, short legs, and thick
skin. They have two functional and two nonfunctional digits on their hooves. Pigs are found and raised all over the world,
providing humans with a variety of products such as pork, lard, leather, glue, fertilizer, and a variety of medicines. Pigs are
most commonly raised in the United States because they produce more lean meat than lard, a cooking fat. (Pig |
Description, Breeds, & Facts, n.d.) Pigs play an important role in ecosystem management and biodiversity conservation.
They create areas for new plant colonization by rooting and thus disturbing the soil. They also disperse fruit seeds, which
helps to spread the plants. Pigs also portray an important role in agricultural resources and animal models in biomedical
studies. Pigs are a useful large animal model because of their advantages over other animals. Pigs have a higher degree
of similarity to humans than rodent models in terms of body/organ size, lifespan, anatomy, physiology, and metabolic
profile. (Genetically Engineered Pigs as Models for Human Disease, 2018) Going back to Nitrogen and phosphorus,
These are nutrients that are found in aquatic ecosystems naturally. The most abundant element in the air we breathe is
nitrogen. Nitrogen and phosphorus help algae and aquatic plants grow, which provide food and habitat for fish, shellfish,
and other aquatic organisms. However, when too much nitrogen and phosphorus enter the environment - which can
happen due to a variety of human activities - the air and water become polluted. For decades, nutrient pollution has
wreaked havoc on many streams, rivers, lakes, bays, and coastal waters, causing serious environmental and human
health problems as well as economic damage. (The Issue, 2021)

Zhenfang Wu and colleagues from South China Agricultural University and other institutions in China, Canada,
and the United States – including joint first authors Xianwei Zhang and Zicong Li – report that genetically engineered pigs
that release fewer of these nutrients could be a solution to the problem.(Transgenic Pigs to the Rescue, 2018) Domestic
animals can now be genetically modified using a variety of techniques. Mammalian genetic modification began with a
pronuclear injection. In essence, a DNA construct was injected into one of the zygote's pronuclei. This technique was first
developed in mice, but it was later applied to a wide range of mammals. Gene editing is a technology that allows and
gives scientists the ability to change the DNA of an individual. This technology allows genetic material to be added,
removed, or altered at a specific location of a genome. (What Are Genome Editing and CRISPR-Cas9?: MedlinePlus
Genetics, n.d.) The use of recently developed gene editing tools to efficiently and precisely modify the pig genome has
greatly expanded the use of pig models in various research areas. Zinc-finger nucleases, transcription activator-like
effector nucleases, and clustered regularly interspaced short palindromic repeats/CRISPR-associated proteins are the
three main types of site-specific nucleases that can efficiently introduce genome modifications, such as knockouts and
knockins, into the pig genome. (Yang, 2018) Pigs can be given desired phenotypes to improve production traits such as
optimal meat production, improved feed digestibility, and disease resistance through these modifications. In 1999, a team
of three researchers led by Dr. Cecil W. Forsberg, Professor Emeritus in Molecular and Cellular Biology, developed the
first phytase pig, dubbed "Wayne." The University holds patents on the technology in the United States and China. The
University of Guelph is entitled to license money and is searching for firms to establish licensing agreements, particularly
in China. This piglet was different from the rest of his litter because he had the phytase gene, a bacterial protein, attached
to a piece of mouse DNA that was locked into one of his chromosomes. This invention is said to help change, improve,
and save lives that everyone will benefit from, however with every breakthrough comes a corresponding risk to many.

DESCRIPTION OF THE TECHNIQUE AND/OR PRODUCT

Pigs' cells, like every other animal, require phosphorus to make DNA, build cell membranes, and transport energy.
The phosphorus element plays an important role in many cellular and organ functions. Domesticated pigs get their daily
dose of corn or cereal grains, but not without a struggle. (CBS News, 2010) Pigs cannot digest a type of phosphorus
found in the foods they eat, therefore farmers supplement their pigs' diets with an enzyme called phytase to help them
break down and digest the phosphorus. Phytase is an enzyme that converts phytate to usable phosphate. Nitrogen and
phosphorus are found naturally in aquatic ecosystems, where they help algae and aquatic plants grow. However, large
amounts of these nutrients can promote the growth of green and blue algae in the environment, particularly in streams,
rivers, bays, and coastal waters. These algal blooms deplete the oxygen in the water, causing aquatic life to perish. Some
algal blooms also produce toxins and promote the growth of bacteria, which can be harmful to people and animals who
come into contact with contaminated water or water that contains too much phosphorus and nitrogen. Human health can
be harmed by nitrogen and phosphorus pollution. Nitrogen-based compounds such as nitrate and nitrite frequently seep
into groundwater in rural areas, posing a health risk to children and pregnant women if consumed through drinking water.
Nitrates cause methemoglobinemia, or 'blue baby' disease, in infants, by preventing the blood from efficiently carrying
oxygen to the organs.(Transgenic Pigs to the Rescue, n.d.) Pigs excrete nearly all of the phosphorus they consume,

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which washes into the ocean and feeds bacteria and algae that cause oxygen "dead zones," which are a major killer of
marine life. In order to fix this problem and prevent it from causing more damage to Earth, scientists came up with a
solution to make the pig produce its own phytase in its salivary glands using the swine's genes When cereal grains are
consumed, they combine with phytase in the pig's saliva, and the enzyme works to break down the phosphorus in the
food throughout the digestive tract. They have invented the ‘Enviropig’. The Enviropig is the world's first pig (a Yorkshire,
to be precise) capable of digesting phytate on its own. (Carey, 2021). This allows the pigs, who are now in their eighth
generation, to get their phosphate solely from grains, excreting about 40% less phosphate. One way to control pollution is
for 'Enviropig' scientists to reduce the levels of nitrogen and phosphorus in swine manure. Pigs excrete large amounts of
these chemicals, owing to their inability to digest phytates and non-starch polysaccharides, two types of molecules found
in their feed. This means that up to 70% of the phosphorus fed to an adult pig will be excreted in the form of bodily waste.
Furthermore, because pigs are unable to digest phytates or non-starch polysaccharides, they are unable to access many
of the nutrients in their feed, limiting their energy intake. The goal of GM Enviropig was to create a product that would
reduce the amount of polluting phosphorus produced by intensive livestock operations or factory farms.

The team that invented and studied the Enviropigs paired the E. coli genes with a mouse DNA promoter, a section
of DNA that encourages replication of a specific segment. In this case, it is the bacterial genes in order to ensure that the
modification would work in mammals. The mixture was then injected into microscopic fertilized pig embryos. Early tests
revealed that the bacterial enzyme was not only incorporated into the pig genome but that it could also be passed down to
the offspring of genetically modified pigs. With these, Enviropig is now capable of absorbing more phosphorus from its
feed, which would result in less of the element being wasted and excreted. ”Pork producers are in favor of any
technologies that can increase their competitiveness,” (P.Sundberg, n.d).

Enviropig has its own advantages and disadvantages. Canada's hog farming industry is in a serious economic
crisis. For some years, farmers have been losing money on every hog they sell, relying solely on off-farm work and
government subsidies. Management of phosphorus overproduction is another expense of production, but the GM pig will
also cost something. Companies may charge a fee per hog, similar to how Monsanto charges a price per acre for the
usage of its proprietary GM crops. Rather than assisting hog growers in dealing with economic challenges, Enviropig
threatens to increase prices while severely undermining domestic and export customer confidence in Canadian pork
products. More than any other country, Canada's hog producers rely on export sales. Canada is the world's third-largest
pork exporter, accounting for 20% of global pork commerce. In contrast, the phytase feed additive is currently inexpensive
and being used by intensive livestock farms as a quick technical fix to fulfill phosphorus pollution requirements. On the
other hand, it can also help farmers. According to National Geographic, Enviropig has the ability to help farmers comply
with the so-called “zero discharge” rules in the country United States, this allows no nitrogen or phosphorus runoff from
the animal operation. (Minard, 2021)

CONTROVERSIES SURROUNDING THE TECHNIQUE AND/OR PRODUCT

With the information above being said, there are some controversies that the implementation of Enviropig. The
following are the factors that led Enviropig to be shut down: Firstly, switching to Enviropigs will be expensive for farmers
since they rely mostly on off-farm work and government subsidies. The process of creating a genetically modified pig or
Enviropig costs an extortionate amount due to the fact that this procedure is new and can only be done by the experts --
with that, we can say that it really is expensive just looking at the people who do this. This procedure also requires years
of study, research, and trial and error in order to serve reliable measures that will ensure the safety of the living things that
require this genome editing. Secondly, for FDA approval, drug makers must undertake lab, animal, and human clinical
testing and submit their results to the FDA in order to obtain FDA clearance. The FDA will then evaluate the data and may
approve the medication if it finds that the drug's benefits exceed the dangers for its intended usage. In Enviropig’s case,
they were not given FDA approval. Lastly, the number of people that disagree with this experiment. Ontario Pork, a hog
business association, has ceased supporting GM pig research at the University of Guelph in Ontario. The institution has
now ceased active research and has terminated its GM pig breeding program. To decrease phosphorus in its feces, the
pig was genetically modified using mouse genetic material. It might have been the world's first GM food animal to be
authorized. This happened in April 2012, because no other source of funding could be identified, the surviving pigs were
killed. However, the Animal Genetic Resources Program in Saskatoon has frozen and preserved Enviropig DNA samples.
(“EnviropigTM” Background, 2019)

These controversies have led the university where the study on the Enviropigs was conducted to close and stop
the study due to the fact that there is no one to fund them. “I had the feeling in seven or eight or nine years that transgenic
animals probably would be acceptable. But I was wrong. It’s time to stop the program until the rest of the world catches
up”. (Forsberg, 2013) Maybe this invention is meant to be invented in the future when people are more open.

LATEST DEVELOPMENT ON THE TECHNIQUE AND/OR PRODUCT

Genome editing comes with a big risk. As said above, the Enviropig was shut down due to a lack of funding
money which led to the euthanasia of the pigs that survived. However, A team of researchers at the University of Guelph
has created a new strain of transgenic "Enviropigs." The Cassie line is the name of the new pig line. Cassie line has the
same ability to digest high levels of phosphorus in plant matter. (Scientists Improve Transgenic “Enviropigs,” n.d.) It is said
that Fifty to seventy percent of the phosphorus in grain is in the form of phytic acid, a molecule that pigs cannot digest. As
a result, many farmers must supplement pig diets with an enzyme known as phytase. Its goal is to break down nutrients to
help pigs digest more nutrients. The transgenic pig produces phytase in its salivary glands, obviating the requirement for
extra supplements or enzymes in the diet. This transgenic is made up of the Escherichia coli phytase coding sequence,

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which is controlled by the mouse parotid secretory protein promoter to drive phytase production in the saliva. The
transgene inserted in the chromosome is present in the CA line as a concatemer of three copies, two in a head-to-tail
orientation and one in a reverse orientation.

However research on the Cassie line has come to an end, researchers have gathered sperm from the pigs and
will be able to produce future Enviropigs. By this, once we will be needing them, we can easily have them. It is also
important because this can be a solution to a problem in the future.

PERSONAL REFLECTION
Genetical engineering indeed has many benefits. One such includes the invention of genetically modified pigs
invented in 1999, the ‘Enviropigs’, studying this made me more knowledgeable about pigs and I also got to know their
function and why is it really important for us, humans, to be able to preserve them. However, pigs are one of the main
contributors to the water pollution that causes marine life to end and perish. This problem should be addressed by the
officials since these two -- pigs and marine life helps us in many big ways -- one of these is pigs being one of the ‘guinea
pig’ or the living things that undergo clinical trial risking their lives in order to save numbers of lives since their internal
organs are the same as a human’s. Indeed, there are many risk factors with genome editing as well as ethical problems,
however, if it has the ability to save the Earth as well as the people who live in it, why not take the risk? We should all see
the bigger picture that this ‘Enviropig’ will give us.

I believe that sometime in the future, once people are finally open, they will be helpful. Contributing to 71% of
water to Earth's surface is contributing to the betterment of the world. The intention of this genetically engineered pig is
good in that it has the goal for marine life to survive. Therefore, I believe that this research should continue.

REFERENCES (APA CITATION STYLE):

Carey, B. (2021, April 26). Enviropig: A Bioengineered Pig That Excretes Fewer Pollutants. Popular Science.
https://www.popsci.com/science/article/2010-09/greener-bacon/

CBS News. (2010, April 5). Genetically Engineered Pigs, Earth-Friendly Poop.
https://www.cbsnews.com/news/genetically-engineered-pigs-earth-friendly-poop/

Genetically engineered pigs as models for human disease. (2018, January 1). PubMed Central (PMC).
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5818075/

Grens, K. (2018, July 31). The Superpowers of Genetically Modified Pigs. The Scientist Magazine®.
https://www.the-scientist.com/notebook/the-superpowers-of-genetically-modified-pigs-64513

The Issue. (2021, August 31). US EPA. https://www.epa.gov/nutrientpollution/issue

Minard, A. (2021, May 3). Gene-Altered “Enviropig” to Reduce Dead Zones? Culture.
https://www.nationalgeographic.com/culture/article/100330-bacon-pigs-enviropig-dead-zones

pig | Description, Breeds, & Facts. (n.d.). Encyclopedia Britannica. Retrieved October 8, 2021, from
https://www.britannica.com/animal/pig-mammal-group

Pollack, A. (2012, April 4). Gene-Altered Pig Project in Canada Is Halted. The New York Times.
https://www.nytimes.com/2012/04/04/science/gene-altered-pig-project-in-canada-is-halted.html

Scientists improve transgenic “Enviropigs.” (n.d.). ScienceDaily. Retrieved October 8, 2021, from
https://www.sciencedaily.com/releases/2013/03/130307124802.htm

Transgenic pigs to the rescue. (n.d.). PubMed Central (PMC). Retrieved October 8, 2021, from
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5963916/#bib9

Yang, H. (2018). Genome Editing of Pigs for Agriculture and Biomedicine. Frontiers.
https://www.frontiersin.org/articles/10.3389/fgene.2018.00360/full

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