Introduction to Biotechnology –scope and importance in fisheries/ aquaculture

Biotechnology arose from the field of zymotechnology, which began as a search for a better
understanding of industrial fermentation, particularly beer.

The heyday and expansion of zymotechnology came in World War I in response to industrial needs
to support the war.

The industrial potential of fermentation was outgrowing its traditional home in brewing, and
"zymotechnology" soon gave way to"biotechnology."

1.1.2 Concepts and Terminologies

The terms biotechnology, genetic engineering and molecular biology are very much related. There
are also many terminologies that have been added to the lexicon of biotechnology. A brif on these
terminologies are summarized hereunder.

[Link]. Biotechnology

It is the application of scientific and engineering principles to the processing of materials by

biological agents to provide goods and services. It involves the use of microorganisms, such as
bacteria or yeasts, or biological substances, such as enzymes, to perform specific industrial or
manufacturing processes. Using bacteria that feed on hydrocarbons to clean up an oil spill is one
example of biotechnology.

[Link]. Genetic engineering

It is also called genetic modification, is the direct human manipulation of an organism's genome
using modern DNA technology. It involves the introduction of foreign DNA or synthetic genes
into the organism of interest.

[Link]. Molecular biology

It is the branch of biology that deals with the formation, structure, and function of macromolecules
essential to life, such as nucleic acids and proteins, and especially with their role in cell replication
and the transmission of genetic information.

[Link]. Genomics

Genomics is the study of the genome . The term genome refers to the entire gene tic content of an
organism. Genomics is the scientific discipline of mapping, sequencing, and analyzing genomes.
The entire RNA and proteincontent of an organism are referred to as transcriptome and proteome,
respectively. Genomics, in the broad sense, includes transcriptomics (study of the transcriptome)
and proteomics (study of the proteome), as the genetic signal can be modified during and after
the transcription and translation processes.
[Link]. Functional genomics
Functional genomics combine bioinformatics, DNA chip technology, animal models, and other
methodologies to identify and characterize genes that cause human disease, and are therefore prime
targets fir drug development.

[Link]. Metagenomics
Metagenomics is the cloning of genetic material from microorganisms that cannot be grown in the
laboratory into ones that can be grown so that new forms of known genes may be identified.

[Link]. Proteomics

The proteome is defined as the expressed protein complement of a cell, tissue, or whole organism.
Proteomics was first used in 1994 by Williams and Hochstrasser. The proteome, unlike the
genome, varies both temporally and between tissues as the fish grows and adapts its physiology to
meet the demands of a new environment. As proteins are the final determinant of phenotype—the
proteome that describes the abundance, identity, posttranslational modifications, and potentially
the synthesis rates of proteins—an understanding of the regulation proteome is imperative to gain
a holistic view of the animal. Proteomics use mass spectroscopy (MS) techniques to identify novel
functional proteins from genes that are expressed.

[Link]. Metabolomics

Metabolomics is the scientific study of chemical processes involving metabolites. Specifically,

metabolomics is the "systematic study of the unique chemical fingerprints that specific cellular
processes leave behind", the study of their small-molecule metabolite profiles. The metabolome
represents the collection of all metabolites in a biological cell, tissue, organ or organism, which
are the end products of cellular processes.

1.1.3. Fields of Biotechnology

Biotechnology has wide applications in all fields of science and technology. Genetic engineering is
used in the production of drugs, human gene therapy , and the development of improved plants.
Based on the field in which the principles of biotechnology are applied, new areas of research have
emerged and have developed into a multi-dicipplinary domains of their own. A brief on such
specialized fields are summarized hereunder.

[Link]. Agricultural biotechnology

Agricultural biotechnology is dated back to 10,000 BC when farmers began to select the most
suitable plants and animals for breeding. Soon thereafter, Sumerians used yeast, a type of fungus,
to make beer and wine in Mesopotamia. In the 1860s, Gregor Mendel crossed different pea plants
and identified the principles of inheritanceand marked the beginning of conventional
biotechnology. Major advances in plant breeding followed the revelation of Mendel’s discovery.
Breeders brought their new understanding of gene tics to the traditional techniques of self-
pollinating and cross-pollinating plants.
Recognising desirable traits and incorporating them into future generations is very important in
plant breeding. A few of these traits can arise spontaneously through a process called mutation,
but the natural rate of mutation is very slow and unreliable to produce all plants that breeders are
looking for. In the late 1920s it was discovered that exposing plants to x-rays and
chemicals could increase the rate of genetic variation, thereby increasing the pool of characteristics
that breeders and farmers could choose from when looking for beneficial features for crop
breeding. Examples of plants that were produced via mutation breeding include varieties of wheat,
barley, rice, potatoes, soybeans and onions.

Experts in United States anticipate the world’s population in 2050 to be approximately 8.7 billion
persons. The world’s population is growing, but its surface area is not. By increasing crop yields,
through theuse of biotechnology the constant need to clear more land for growing food is reduced.

Countries in Asia, Africa, and elsewhere are grappling with how to continue feeding a growing
[Link] are also trying to benefit more from their existing [Link] holds
the key to increasing the yield of staple crops by allowing farmers to reap bigger harvests from
currently cultivated land, whilepreserving the land’s ability to support continued farming.

Malnutrition in underdeveloped countries is also being combated with biotechnology. The

Rockefeller Foundation is sponsoring research on “golden rice”, a crop designed to improve
nutrition in the developing world. Rice breeders are using biotechnology to build Vitamin A into
the rice. Vitamin A deficiency is a common problem in poor countries. A second phase of
theproject will increase the iron content in rice to combat anemia, which is a widespread problem
among women and children in underdeveloped countries.

Similar initiatives using gene tic manipulation are aimed atmaking crops more productive by
reducing their dependence on pesticides, fertilizers and irrigation, or by increasing their resistance
to plant diseases. Increased crop yield, greater flexibility in growing environments, less use of
chemical pesticides and improved nutritional content make agricultural biotechnology, quite
literally, the future of the world’s foodsupply.

The plant biotechnology has following applications:

1. Plant Cell and Tissue Culture .

2. Production of pesticide, herbicide and salt tolerant plants. For example, an “insect protection”
gene (Bt) has been inserted into several crops - corn, cotton, and potatoes - to give farmers new
tools for integrated pest management. Bt corn is resistant to European corn borer. This inherent
resistance thus reduces a farmers pesticide use for controlling European corn borer, and in turn
requires less chemicals and potentially provides higher yielding Agricultural Biotechnology.

[Link]. Animal biotechnology

The animal biotechnology has the following application.

1. The development of vaccines to protect animals from disease,

2. The production superior calves through superovulation and embryotranfer technology,

3. The production of several calves from one embryo (embryo splitting/ cloning ),

4. Increase of animal growth rate,

5. Rapid disease detection by molecular immunological techniques.

6. Monoclonal antibody production

7. Recombinant vaccine production, and

8. Transgenic technology in animal production.

In summary, modern biotechnology offers opportunities to improve product quality, nutritional

content, and economic benefits. The genetic makeup of plants and animals can be modified by
either insertion of new useful genes or removal of unwanted ones. Biotechnology is changing the
way plants and animals are grown, boosting their value to growers, processors, and consumers.

[Link]. Industrial Biotechnology

Industrial biotechnology applies the techniques of modern molecular biology to improve
the efficiency and reduce the environmental impacts of industrial processes like textile, paper and
pulp, and chemical manufacturing. For example, industrial biotechnology companies develop
biocatalysts, such as enzymes, to synthesize chemicals. Enzymes are proteins produced by all
organisms. Using biotechnology, the desired enzyme can be manufactured in commercial
quantities.
Biotechnology also produces biotech-derived cotton that is warmer, stronger, has improved
dye uptake and retention, enhanced absorbency, and wrinkle- and shrink-resistance.
Some agricultural crops, such as corn, can be used in place of petroleum to produce
chemicals. The crop’s sugar can be fermented to acid, which can be then used as an intermediate
to produce other chemical feedstocks for various products. It has been projected that 30% of the
world’s chemical and fuel needs could be supplied by such renewable resources in the first half of
the next century.

[Link]. Environmental Biotechnology

Environmental biotechnology is used in waste treatment and pollution prevention.
Environmental biotechnology can more efficiently clean up many wastes than conventional
methods and greatly reduce our dependence on methods for land-based disposal.
Every organism ingests nutrients to live and produces by-products as a result. Different
organisms need different types of nutrients. Some bacteria thrive on the chemical components of
waste products. Environmental engineers use bioremediation, the broadest application of
environmental biotechnology, in two basic ways. They introduce nutrients to stimulate the activity
of bacteria already present in the soil at a waste site, or add new bacteria to the soil. The bacteria
digest the waste at the site and turn it into harmless byproducts. After the bacteria consume the
waste materials, they die off or return to their normal population levels in the environment.
Bioremediation, is an area of increasing interest. Through application of biotechnical
methods, enzymebioreactors are being developed that will pretreat some industrial waste and food
waste components and allow their removal through the sewage system rather than through solid
waste disposal mechanisms. Waste can also be converted to biofuel to run generators. Microbes
can be induced to produce enzymes needed to convert plant and vegetable materials into building
blocks for biodegradable plastics.
In some cases, the byproducts of the pollution-fighting microorganisms are themselves
useful. For example, methane can be derived from a form of bacteria that degrades sulfur liquor, a
waste product of paper manufacturing. This methane can then be used as a fuel or in other industrial
processes.

[Link]. Fisheries Biotechnology

Biotechnology is also used in the fisheries field for increasing fish production through various
techniques. Fisheries biotechnology can be broadly classified into aquaculture biotechnology,
marine biotechnology, algal biotechnology and processing biotechnology.

Since 1980s, there has been a burst of biotechnology activity in research and development related
to various fish species, in particular those used in aquaculture production. Biotechnology has
played a major role in the areas of induction and control of

maturation and spawning, sex control (andro gene sis and gynogenesis), sex inversion in
protandrous species like sea bass and protogynous species like the grouper, production of triploid,
tetraploid and transgenic fishes. Traits that are being tested in fish species such as carp, trout,
salmon and channel catfish include growth rates that are three to eleven times faster with
more efficient feed utilisation, increased tolerance to cold water and improved disease resistance.
Accelerated growth rates mean that fish reach marketable size sooner, thereby reducing overhead
costs for fish farmers. In addition, researchers use the human interferon gene to improve disease
resistance in carp, which could reduce the amount of antibiotic s needed to keep fish healthy and
reduce the costs incurred from losses due to disease.

The first (and to date only) genetically engineered fish to be sold commercially is the fluorescent
Glofish®, a zebra fish modified to glow red, which came onto the US market in 2004.

Other areas include disease diagnosis (molecular and immunodiagnostic

kits), hybridoma technology, and management (probiotics, vaccines, immunostimulants), cell
and tissue culture , conservation of germplasm (cryopreservation of fish gametes), extraction of
bioactive substances from marine organisms including marine bacteria, marine algae, marine
invertebrates and fishes.

[Link]. Other Applications

Biotechnical methods are now used to produce many protein s for pharmaceutical and other
specialized purposes. A harmless strain of Escherichia coli bacteria, given a copyof the gene for
human insulin, can make insulin. As these genetically modified (GM) bacterial cells age, they
produce human insulin, which can be purified andused to treat diabetes in humans. Products of
modern biotechnology include artificial blood vessels from collagen tubes coated with a layer of
theanticoagulant heparin.

Gene therapy – altering DNA within cells in an organism totreat or cure a disease – is one of the
most promising areas of biotechnologyresearch. New genetic therapies are being developed to treat
diseases such ascystic fibrosis, AIDS and cancer.

DNA fingerprinting has become one of the most powerful andwidely known applications of
biotechnology today. DNA from samples of hair, bodily fluids or skin at a crime scene are
compared with those obtained fromthe suspects.

1.1.4. Historical events related to biotechnology

The Hungarian Karl Ereky coined the word "biotechnology" in 1919 to describe a technology
based on converting raw materials into a more useful product. For Ereky, the term "biotechnologie"
indicated the process by which raw materials could be biologically upgraded into socially useful
products.

• In1920, Leads city council, U.K. established the Institute of Biotechnology.

• During1970s, Biotechnology emerged as a new discipline.
• In 1978, European Federation of Biotechnology was established.

Biotechnology is the application of scientific and engineering principles to the processing of

materials by biological agents to provide goods and service (The Organisation for Economic Co-
operation and Development, OECD, 1981). The "Scientific and Engineering Principles" refer to
microbiology, genetics,

• biochemistry,etc. and "biological agents" mean microorganisms, enzymes, plant and

animal cells.
• In 1982, Government of India set up, the National Biotechnology Board and in 1986, it be
came a separate department, Department of Biotechnology in the Ministry of Science and
Technology.
• United Nations proposed for the establishment of International Centre for Genetic
Engineering and Biotechnology (ICGEB) in 1988. It has 2 centres, New Delhi (India) and
Trietse.
• In the 1940s, penicillin was discovered in England and it was produced industrially in the
United States using a deep fermentation process. The enormous profits and the public
expectations penicillin gave rise to a radical shift in the standing of the pharmaceutical
industry. Doctors used the phrase "miracle drug".
• A number of discoveries made during the 1960s and 1970s shed light on how distinct
fragments of DNA could be isolated .
• The work of Swiss molecular biologist Werner Arber focused on specialized enzymes that
digest, or “restrict,” the DNA of viruses infecting bacteria. These enzymes were
subsequently called as “ restriction enzyme s” that could also act like molecular scissors to
cut DNA.
• In 1970 American molecular biologist Hamilton Smith and colleagues determined that
restriction enzymes could cleave DNA molecules at precise and predictable locations.
Hamilton concluded that the enzymes were able to recognize
specific nucleotide sequence s. Scientists quickly realized that restriction enzymes could
be used in the laboratory to manipulate DNA.
• In 1973 American biochemist Herb Boyer used restriction enzymes to produce a DNA
molecule with genetic material from two different sources. This splicing technique is now
known as recombinant DNA .
• Boyer inserted foreign genes into plasmid s and observed that the plasmids could replicate
to make many copies of the inserted genes. In subsequent experiments, Boyer, American
biochemist Stanley Cohen, and other researchers demonstrated that inserting a recombinant
DNA molecule into a host bacteria cell would lead to extremely rapid replication and the
production of many identical copies of there combinant DNA.
• This process, known as cloning , gave scientists the power to make many copies of desired
DNA for molecular study.
• The speed and efficiency of DNA cloning were vastly improved in the 1980s with the
invention of polymerasechain reaction (PCR). Developed by American biochemist Kary
Mullis , PCR enables scientists to produce large amounts of DNA sequence s ina test tube.
In a matter of hours, the process can produce millions of cloned DNA molecules.
• In the late 1970s and early 1980s, British biochemist Frederick Sanger and his associates
developed DNA sequencing techniques . Sanger’s methods, which used special
compounds called dideoxy nucleotides, rapidly yielded the exact nucleotide sequence of a
desired sample. With the use of automated equipment, the new techniques transformed
genetic sequencing into a speedy, routine laboratory procedure.
• [Link]. Other significant events
• 6000B.C- Bread making (involving yeast fermentation)
• 1857AD- Pasteur proves that yeasts are living cells that cause alcohol fermentation
• 1928– Alexander Fleming discovers penicillin from Penicillium notatum
• 1953-DNA structure and function elucidated
• 1970-Smith et al. report restriction endonuclease from Haemophilus influenzae that
recognizes specific DNA targetsequence s
• 1972- Walter Fiers and his team at the Laboratory of Molecular Biology of the University
of Ghent ( Ghent ,Belgium ) were the first to determine the sequence of a gene : the gene
for bacteriophage MS2 coat protein .
• 1973- Tong et al. injected mRNA and rRNA from mature eggs of crucian carp and
common carp into newly fertilized crucian carp eggs, in order to induce character variation
in goldfish.
• 1973- Tong and Niu, transplantednuclei between gold fish (Carassiusauratus)
and Rhodeus sinensis forthe purpose of studying the developmental variations between the
integratednuclei and the pure heterologous nuclei, and the effects of cytoplasm on
thenucleus.
• 1974- Parker defined probiotics are “organisms and substances which contributeto
intestinal microbial balance”.
• 1976 - Walter Fiers andhis team determine the complete nucleotide -sequence
of bacteriophage MS2-RNA
• 1975-Kohler and Milstein report monoclonal antibodies
• 1977- DNA is sequenced forthe first time by Fred Sanger , Walter Gilbert , and Allan
Maxamworkingindependently. Sanger's lab sequence theentire genome of bacteriophage
Φ-X174 .
• 1979- Paulien Hogeweg coined the term bioinformatics for the study of informatic
processesin biotic systems.
• 1980 - Gordon etal. revolutionized the procedure for producing transgenic animals based
onthe microinjection ofclone d DNA into the pronucleus of fertilized eggs at theone-cell
stage.
• 1982 - Induced the first viable tetraploid fish, rainbow trout
• 1982-Palmiter et al., produced firsttransgenic mouse; rat gene transfer red to mouse
• 1983 - Kary B. Mullis discoversthe polymerase chainreaction enabling theeasy
amplification of DNA
• 1984-Transgenic pig, rabbit, and sheep by microinjection of foreign DNA into eggnuclei
• 1984- Maclean and Talwar reported microinjection of cloned DNA intorainbow trout
(Oncorhynchus mykiss)eggs.
• 1985-First transgenic fish was produced, Zhu produced transgenic goldfish
• 1986 - Fletcher et al.,showed that AFP injection to seawater-acclimatized rainbow trout
lowered thefreezing point of the whole fish in proportion to the circulating anti-
freezeprotein concentration.
• 1986- Chen et [Link] cell nuclei from a grass carp blastula cell line
intounfertilized, enucleated eggs of crucian carp, thus creating the first "test-tube fish".
• 1989-The human gene that encodes the CFTR proteinwas sequenced by Francis
Collins and Lap-Chee Tsui . Defects in this gene cause cystic fibrosis
• Human Genome Project started
• 1990- Shujian et al. transplanted cell nuclei of themutant cell line (AHZC- 88), which
• was resistant to the grass carphemorrhagic virus, into unfertilized grass carp eggs
• using electric fusion, and raisedthree of the fish to the fry stage.
• 1995-The genome of Haemophilusinfluenzae is the first genome of a free living organism
to be sequenced.
• 1996- Saccharomyces cerevisiae is the first eukaryote genomesequence to be released.
• 1998-The first genome sequence for a multicellular eukaryote, Caenorhabditis elegans , is
released.
• 1999- Zhiyuan Gong et al., at the National University of Singapore produced transgenic
Zebrafish by inserting a gene called green fluorescent protein (GFP), originally extracted
from a jellyfish , that naturally produced brightgreen bioluminescence .
• 2001-First draft sequences of the human genome are released simultaneously by
the Human Genome Project andCelera Genomics .
• 2003- Successful completion of Human Genome Project with 99% of the
genomesequenced to a 99.99% accuracy .
• 2003- Gong et al., developed transgenic zebrafish (Danio rerio) for ornamental
andbioreactor system by strong expression of fluorescent proteins in the skeletalmuscle.