NAME: BRENDA ATIENO
SCHOOL: PURE AND APPLIED SCIENCE
DEPARTMENT: BIOCHEMISTRY
ADM NO. : I28/4889/2021
UNIT TITLE. : BIOSAFETY AND BIOETHICS
UNIT CODE. : SBC 447
SUBJECT. : ASSIGNMENT
[Link] Issues raised about GMO
Genetic modification is a special set of gene technology that alters the genetic machinery of such
living organisms as animals, plants or microorganisms. Combining genes from different
organisms is known as recombinant DNA technology and the resulting organism is said to be
‘Genetically modified (GM)’, ‘Genetically engineered’ or ‘Transgenic’.
The biggest threat caused by GM foods is that they can have harmful effects on the human body.
It is believed that consumption of these genetically engineered foods can cause the development
of diseases which are immune to antibiotics. Besides, as these foods are new inventions, not
much is known about their long term effects on human beings. As the health effects are
unknown, many people prefer to stay away from these foods. Manufacturers do not mention on
the label that foods are developed by genetic manipulation because they think that this would
affect their business, which is not a good practice. Many religious and cultural communities are
against such foods because they see it as an unnatural way of producing foods. Many people are
also not comfortable with the idea of transferring animal genes into plants and vice versa. Also,
this cross-pollination method can cause damage to other organisms that thrive in the
environment. Experts are also of the opinion that with the increase of such foods, developing
countries would start depending more on industrial countries because it is likely that the food
production would be controlled by them in the time to come.
Genetic Contamination/Interbreeding Introduced GMOs may interbreed with the wild-type or
sexually compatible relatives. The novel trait may disappear in wild types unless it confers a
selective advantage to the recipient. However, tolerance abilities of wild types may also develop,
thus altering the native species’ ecological relationship and behaviour.
Competition with Natural Species Faster growth of GMOs can enable them to have a
competitive advantage over the native organisms. This may allow them to become invasive, to
spread into new habitats, and cause ecological and economic damage.
Increased Selection Pressure on Target and Nontarget Organisms Pressure may increase on
target and nontarget species to adapt to the introduced changes as If to a geological change or a
natural selection pressure causing them to evolve distinct resistant populations.
Ecosystem Impacts The effects of changes in a single species may extend well beyond to the
ecosystem. Single impacts are always joined by the risk of ecosystem damage and destruction.
Impossibility of Follow up Once the GMOs have been introduced into the environment and
some problems arise, it is impossible to eliminate them. Many of these risks are identical to those
incurred with regards to the introduction of naturally or conventionally bred species. But still this
does not suggest that GMOs are safe or beneficial, nor that they should be less scrutinized.
Horizontal Transfer of Recombinant Genes to Other Microorganisms One risk of particular
concern relating to GMOs is the risk of horizontal gene transfer (HGT). HGT is the acquisition
of foreign genes (via transformation, transduction, and conjugation) by organisms in a variety of
environmental situations. It occurs especially in response to changing environments and provides
organisms, especially prokaryotes, with access to genes other than those that can be inherited
[17, 18].
[Link] is risk assessment of GMO conducted?
Directly observable risks are assessed and managed through heuristic processes. This direct
observation may sometimes be insufficient to establish the nature and extent of risk. In such
cases, we rely on other institutions, especially reputation and the rule of law [35]. Biosafety
issues pertaining to the marketing of GMOs have received increasing attention by national and
international agencies and regulatory bodies worldwide [2, 4, 36]. These are based on a common
set of principles built on the accumulation of experience and scientific knowledge over the past
decades. Risk assessment intends to quantify risks and evaluate the probabilities of possible
outcomes on the basis of scientific data. It is a fundamental part of improving quality, being the
quality of products or the quality of life, and plays a central role in the innovation required to
maximize benefits. A critical step in risk assessment is identification of circumstances that may
give rise to an adverse effect(s) (risk identification or “what could go wrong’’ step) [25]. The
level of risk is then estimated from both the likelihood (“how likely is it to happen’’ step) and
severity/consequences (“would it be a problem’’ step) associated with the circumstances of
concern. This is then followed by characterization of the risk based on evaluation of likelihood
and consequences of the identified adverse effects being realized (“what is the risk’’ step) [11].
The international agreements such as Cartagena Protocol on Biosafety, Convention on Biological
Diversity (CBD), and the International Plant Protection Convention (IPPC) address the
environmental aspects of GMOs. The Article 15 of the Cartagena Protocol on Biosafety (CBD
2000) implies risk assessment to be in compliance with criteria of science and transparency using
already existing and recognized techniques. The characterization process should adopt a
multidisciplinary approach that
(i)
Analyses methodologies in statistics,
(ii)
Considers the individual components employed to produce the GMOs (such as characteristics of
the donor organism, vector, and inserted DNA),
(iii)
Evaluates the final result in its totality (characteristics of the organism with new traits,
information related to intended use, and characteristics of the potential receiving environment),
(iv)
Considers relevant information produced from both public and private research institutes and
from international agencies.
[Link] of adoption of GMO in Kenya
Kenya enacted the Biosafety Act in 2009 and established NBA in 2010. Since its establishment,
NBA has reviewed over 28 contained use applications. These are applications done in the lab.
We have approved 14 confined field trials and reviewed two environmental release applications
for Bt cotton and Bt maize. Bt cotton has moved on and we expect national performance trials to
commence (soon). Bt maize still has some issues to [Link] recently lifted a ban on the
cultivation and importation of genetically modified crops amid the worst drought in 40 years and
soaring food prices. This includes white maize, the country’s main staple. The decision was
welcomed by scientists who see GM crops as the answer for food security.
[Link] of good Biological techniques
Cell Lysis
To separate compounds from cellular environments, one must first break open (lyse) the cells.
Cells are broken open, in buffered solutions, to obtain a lysate. There are several ways of
accomplishing this.
Fractionation and Chromatography Techniques
Fractionation of samples, as the name suggests, is a process of separating out the components or
fractions of the lysate. Fractionation typically begins with centrifugation of the lysate. Using
low-speed centrifugation, one can remove cell debris, leaving a supernatant containing the
contents of the cell. By using successively higher centrifugation speeds (and resulting g forces) it
is possible to separate out different cellular components, like nuclei, mitochondria, etc., from the
cytoplasm.
Electrophoresis
Electrophoresis uses an electric field applied across a gel matrix to separate large molecules such
as DNA, RNA, and proteins by charge and size. Samples are loaded into the wells of a gel matrix
that can separate molecules by size and an electrical field is applied across the gel. This field
causes negatively charged molecules to move towards the positive electrode. The gel matrix,
itself, acts as a sieve, through which the smallest molecules pass rapidly, while longer molecules
are slower-moving.
Detection, identification and quantitation of specific nucleic acids and proteins
One way to detect the presence of a particular nucleic acid or protein is dependent on transferring
the separated molecules from the gels onto a membrane made of nitrocellulose or nylon to create
a “blot” and probing for the molecule(s) of interest using reagents that specifically bind to those
molecules. The next section will discuss how this can be done for nucleic acids as well as for
proteins.
Transcriptomics
Consider a matrix containing all of the known gene sequences in a genome. To make such a
matrix for analysis, one would need to make copies of every gene, either by chemical synthesis
or by using PCR. The strands of the resulting DNAs would then be separated to obtain single-
stranded sequences that could be attached to the chip. Each box of the grid would contain
sequence from one gene. One could analyze the transcriptome – all of the mRNAs being made in
selected cells at a given time.
Isolating Genes
Methods to isolate genes were not available till the 1970s, when the discovery of restriction
enzymes and the invention of molecular cloning provided, for the first time, ways to obtain large
quantities of specific DNA fragments, for study. Although, for purposes of obtaining large
amounts of a specific DNA fragment, molecular cloning has been largely replaced by direct
amplification using the polymerase chain reaction described later, cloned DNAs are still very
useful for a variety of reasons.
Polymerase Chain Reaction (PCR)
The polymerase chain reaction (PCR) allows one to use the power of DNA replication to amplify
DNA enormously in a short period of time. As you know, cells replicate their DNA before they
divide, and in doing so, double the amount of the cell’s DNA. PCR essentially mimics cellular
DNA replication in the test tube, repeatedly copying the target DNA over and over, to produce
large quantities of the desired DNA.
Reverse Transcription
In the central dogma, DNA codes for mRNA, which codes for protein. One known exception to
the central dogma is exhibited by retroviruses. These RNA-encoded viruses have a phase in their
life cycle in which their genomic RNA is converted back to DNA by a virally-encoded enzyme
known as reverse transcriptase. The ability to convert RNA to DNA is a method that is desirable
in the laboratory for numerous reasons.
FRET
The fluorescence resonance energy transfer (FRET) technique is based on the observation that a
molecule excited by the absorption of light can transfer energy to a nearby molecule if the
emission spectrum of the first molecule overlaps with the excitation spectrum of the second.
This transfer of energy can only take place if the two molecules are sufficiently close together
(no more than a few nanometers apart.
Genome Editing (CRISPR)
The development of tools that would allow scientists to make specific, targeted changes in the
genome has been the Holy Grail of molecular biology. An ingenious new tool that is both simple
and effective in making precise changes is poised to revolutionize the field, much as PCR did in
the 1980s. Known as the CRISPR/Cas9 system, and often abbreviated simply as CRISPR, it is
based on a sort of bacterial immune system that allows bacteria to recognize and inactivate viral
invaders.
Protein Cleavage
Because of their large size, intact proteins can be difficult to study using analytical techniques,
such as mass spectrometry. Consequently, it is often desirable to break a large polypeptide down
into smaller pieces. Proteases are enzymes that typically break peptide bonds by binding to
specific amino acid sequences in a protein and catalyzing their hydrolysis.
Membrane Dynamics (FRAP)
Understanding the dynamics of movement in the membranes of cells is the province of the
Fluorescence Recovery After Photobleaching (FRAP) technique. This optical technique is used
to measure the two dimensional lateral diffusion of molecules in thin films, like membranes,
using fluorescently labeled probes. It also has applications in protein binding.
[Link] levels I experienced during attachment
In order to prevent the contamination of personal clothing, protective laboratory coats, gowns, or
uniforms were recommended for me .
While conducting tests with a high possibility of aerosol formation, I used protective eyewear .
The laboratory supervisor implemented the policies regarding the access control to the
laboratory.
Activities like eating, drinking, smoking, handling contact lenses, applying cosmetics, and
storing food were not permitted in laboratory areas.
I used to wash my hands after using viable microorganisms and before leaving the laboratory.
I used to do Mechanical pipetting instead of mouth pipetting.
All contaminated cultures, glassware, plastic ware, and biologically contaminated waste were
treated as bio-hazards and thus, autoclaved.
I used to decontaminate work places with disinfectant at the end of the day and after any spills or
splashes.
I used to treat used hypodermic syringes and needles, Pasteur pipettes, razor blades,
contaminated broken glass, and blood vials are medical waste and discard in puncture-resistant
sharps disposal containers.
I used to wear protective coat while entering the laboratory and then remove and keep in the
laboratory post work.
Some days I used to work in a BSC class 2a
[Link] of Infection
The following are the primary routes of transmission that can result in laboratory acquired
infections.
Injection (percutaneous)
Contaminated sharp objects (e.g. needle, scalpel)
Animal bites, scratches
Through broken or abraded skin (including rashes, eczema, split cuticles, etc.)
Absorption (mucous membrane contact)Routes of infection
Splashes to the eyes, nose, mouth
Hand to face movements (i.e., applying cosmetics, cell phone usage, etc.)
Ingestion
Eating/drinking
Applying cosmetics
Contact with tear ducts
Inhalation (aerosols)
Liquid disturbance
Syringe preparation
Dried animal excretions
Leakage from injection site
The diagram below illustrates different routes of infection
6)Constitution and role of Institutional Biosafety Committee
Functions of Management
Ultimate responsibility is to ensure the working Environment is safe
a) Undertake risk assessment on human health and environmental safety
b) Appoint IBC/BSO to advise on risk assessment
c) Ensure adequate containment facilities
d) SOPs are in place to control any risk to workers
e) Formulate and implement local rules, Emergency plans and procedures
f) Adequate training for the level of risk
Composition of IBC
a) Biosafety officer (s)
b) Scientist (s)in the relevant field
c) Representative (s)of technical staff
d) Representative (s) of lab management
e) Representative of the community
f) Representative (s)of the relevant (RA)
Functions of IBC
a) Facilitate applications preparation
b) Advise institution on Biosafety matters
c) Assist institutions to establish monitoring plan for risk assessment & management
d) Ensure compliance to NBA approval conditions
e)Review and ascertain suitability of containment (Physical & Biological)
f)Advise institution & PI on mitigation measures in case of accident
Biosafety Officer
Overall to secure Biosafety of the Installation
a) Supervise contained use risk evaluation
b) Coordinate notifications and applications related with contained use
c) Train staff & Control GMO storage and transport
d) Manage waste treatment
e) Supervise necessary action in case of accident
f) Ensure traceability of information
g) Organize and participate in internal audits
h) Maintenance & control of equipment
Principal Investigator
Responsible for full compliance
a) Adequately trained in biological techniques
b) Provide Lab. Staff with protocols (potential biohazards and necessary precautions)
c) Instruct and train Lab. Staff on biosafety
d) Inform Lab. Staff reasons for serum collection
e) Supervise Lab. Staff to follow safety practices
f) Integrity of biological & physical containment
g) Comply: permit & shipping condition of GMO
h) Correct work errors and conditions.
References
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effects of GM crops, Proceedings of the Royal Society B: Biological Sciences. (2007) 274, no.
1617, 1475-1479, 2-s2.0-34447320436, [Link]
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33. Losey J. E., Rayor L. S., and Carter M. E., Transgenic pollen harms monarch larvae, Nature.
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