a. As per Balloux and Dorp (2017), the microorganisms that cause diseases to their hosts are termed as pathogenic microorganisms. There are two main types of pathogenic microorganisms that cause severe disorders to their hosts; these are bacteria, and viruses. Bacteria are the single-celled prokaryotes that do not have a well-defined genetic material. Comma-shaped bacteria, rod-shaped bacteria, spiral-shaped bacteria, and spherical-shaped bacteria are the various types of bacteria on the basis of shape. Some bacteria live as single cells, while others live in the form of colonies (Microbiology Society, 2019). Bacteria have an outer cell wall for protection; cilia and flagella that support locomotion. Viruses are the organisms that have an outer protein coat called capsid that contains the genetic material. All viruses require a host organism to reproduce and make their multiple copies (Louten, 2016).
b. The microorganisms that convert themselves into pathogens under certain circumstances are termed as opportunistic pathogens. They attack the hosts that have a suppressed immune system because it is easy to attack a weak immune system than attacking a strong immune system. These types of microorganisms thrive better when the host's body is deteriorating. For example, the patients suffering from AIDS (Acquired Immunodeficiency Syndrome) are easy targets of opportunistic microbes (Biology LibreTexts, 2019a). As per Biology LibreTexts (2019d), opportunistic pathogens differ from primary pathogens in the following ways:
c. All the following general safety rules and regulations must be followed while working with microorganisms in the biomedical laboratories (America Society of Microbiology, 2019):
Always wear laboratory coats, safety goggles, gloves, and cover the head while working in the biomedical labs. This prevents any type of direct contact with the microorganisms and reduces the chances of microbial infections.
Do not bring any personal belonging inside the lab to keep all personal belongings safe from microbial contamination.
All eatable items must be prohibited inside the labs; all the people working with the microbes should not eat anything inside the labs. This is because the microbes present in the biomedical labs can contaminate food and water.
It is advisable that people working in the microbiological labs should not touch their face, or bite their nails as the microbes can enter inside the body through these steps.
Always tie the hairs properly while working in biomedical labs as the hairs can contaminate the microbial culture.
All types of sharp materials must not be used in the labs. Only the needles and other sharp materials that are provided by the institution should be used while working in the labs.
All the disposable items must be thrown properly and safely. For example, wipe the gloves with ethanol before throwing them to avoid the risk of microbial infections.
All these rules and regulations are important while working with microbes as they reduce the risk of direct contact with harmful microorganisms.
d. As per King and Kang (2018), ultraviolet radiations are used to destroy the pathogenic microorganisms such as bacteria and fungi. There are different types of UV rays, such as UV-A, UV-C, and others. However, UV-C is most effective in destroying microorganisms as it destroys their genetic material. The UV-C rays are used in biomedical laboratories to clean the equipment and make them free of unwanted microorganisms. It helps researchers in conducting safe and effective experiments. Most of the bacteria and other organisms die due to UV-C rays, but some are still viable. The number of viable bacteria that are left after incubation with UV-C rays can be counted by the method of plate count. Plate count is one of the most commonly used methods of detecting viable bacteria in a solution. In this method, the number of actively growing bacteria is counted to identify the number of bacteria which are still alive after being irradiated by UV-C rays (Biology LibreTexts, 2019b).
a. As per Louten (2016), viruses are small organisms that need a host to reproduce. These are very small in size and have an outer protein coat that contains deoxyribonucleic acid (RNA). This outer protein coat is termed as the capsid, and it protects the viral genome from the external environment. The nucleic acid, along with the capsid, forms a structure called nucleocapsid. The viral genome is enclosed by the protein coat or capsid. There are different forms of viruses on the basis of structure. For example, the viruses that have helical capsids are termed as helical viruses. The viruses that have an icosahedral capsid are known as icosahedral viruses, while the viruses that have a complex capsid are known as complex viruses. These viruses do not have a helical or icosahedral capsid. The complex viruses are named so because they have complex structures such as protein tails (Biology LibreTexts, 2020e).
b. Viral diagnosis is the method that is used to detect any form of viral infection. There are three types of viral diagnosis; direct detection, serology tests, and viral isolation. The direct examination of viral particles and antigens in the host body is termed as direct detection. In this method, Transmission electron microscopy (TEM) is used to collect the direct images of the viruses. Another method of viral diagnosis is immunofluorescence assay in which the virus antigens and virus-related antibodies are detected to check the presence of viral infections. Immunofluorescence assay is a type of viral isolation method because the viral antigens and virus-related antibodies are isolated in this method. In this method, stains such as fluorescent antibody (FA) stains; immunoperoxidase stains are needed. However, the most common way of viral diagnosis is serology testing, in which the rising titres of antibodies are detected. The amount of antibody IgM is also detected in serology testing because the body releases IgM antibodies during viral attacks. There are different types of serology tests such as Western blotting, particle agglutination, Enzyme-linked immunosorbent assay (ELISA), and many more.
The serology method is used for detecting the virus infections that cause measles, hepatitis A, hepatitis B, and many more (Brahamchari, 2019).
c. As per AFMC (2016), the best way to prevent all types of viral infections is to maintain personal hygiene. It is very important to follow a healthy lifestyle to reduce the risks of viral infections. For example, a healthy diet, regular exercise, personal fitness, drinking clean water, and hygiene are important as they boost up the body's immune system. A person with a healthy immune system is at a lower risk of a viral infection than people with a poor immune system.
a. The cells that are isolated directly from living organisms such as humans and animals is termed as primary cell lines. These are important for the purpose of research. The primary cell lines obtained by the donors are used to understand the biological processes in the in-vitro conditions. These cell lines are used for tissue engineering studies. The primary cell lines are also used for conducting regenerative studies (Weigand et al., 2016).
b. The process of isolating cells from humans or animals and then proliferating them is termed as primary cell culture. In this method, the isolated cells are kept in a solution to induce proliferation. The proliferated cells are then transferred into new media solutions for continued growth. The tissues isolated from the organs can be grown in-vitro by following the steps of primary cell culture. The process of primary cell culture is performed in three major steps. The first step is isolation in which a part of the organ is isolated and removed out by using mechanical and enzymatic dissociation. Trypsinization is one of the most commonly used methods of enzymatic dissociation used for the development of primary cell culture. The second step is culturing in which the isolated tissues are cultured in the culture media. And the final step is the proliferation in which the cultured tissues are added in new growth media to induce continued growth (Bhatia, Naved & Sardana, 2019).
c. Leonard Hayflick was the scientist who introduced the concept of Hayflick phenomena and related it with cellular ageing. This phenomenon helped the scientist in understanding the effects of cellular ageing from birth till death. Hayflick phenomena or Hayflick limit is defined as the number of times human cells divide before the termination of cell division (Ndifon & Dushoff, 2016).
d. Stem cells differ from the differentiated cells in the following ways (Lakna, 2017):
a. Deoxyribonucleic acid (DNA) is a long, double-stranded molecule that can be cut into smaller fragments by using restriction enzymes. The enzymes that are used to cleave DNA is termed are termed as restriction enzymes. These enzymes are of two types, endonucleases and exonucleases. The restriction enzymes that cut the DNA at the terminal ends are called exonucleases enzymes, while the ones that cleave the DNA within the strand are called endonucleases. The restriction enzymes are highly specific and cleave the DNA only at specific reorganization sites. Different restrictions enzymes are available to cut the DNA; for example, EcoRI and HindIII. The process of separating the DNA fragments based on their molecular weight is termed as gel electrophoresis. Agarose gel is used to separate the DNA fragments as it has very few charged particles. It is also very easily cast, and the DNA sample that has DNA fragments are loaded in this agarose gel. The DNA fragments start moving in the agarose gel, and their movement is based upon their molecular weight. The DNA fragments that have low molecular weight travel faster while the ones with heavy molecular weight move slower. The DNA fragments are stained with a solution of Ethidium bromide (EtBr) for the process of visualization. The DNA fragments that are stained with EtBr appear orange when placed in the UV box. They reflect an orange color in the presence of UV light. In this way, the separated DNA fragments are visualized after gel electrophoresis (Biology LibreTexts, 2019c).
b. As per Science Learning Hub (2017), PCR or Polymerase Chain Reaction is defined as the molecular biology technique which is used for DNA amplification. It is used by the researchers to amplify the DNA sample. The process of PCR helps the scientists and researchers to make many copies of the small DNA sample. The method of PCR is important in research and medical diagnosis as it is difficult to conduct long term researches by using a small DNA sample. Researchers need a sufficient amount of DNA samples to conduct complex experiments. The basic requirements of PCR are the DNA segment that is to be amplified, primers, nucleotides, restriction enzymes, and Taq polymerase. The process of PCR starts with denaturation, in which the two strands of DNA are separated from each other. The next process is known as annealing, in which primers are added to the DNA to induce polymerization. The final step is termed as an extension in which Taq polymerase and nucleotides are added to the DNA to increase its length and to form its copies. The results of PCR can be viewed by using the method of electrophoresis. The newly produced DNA fragments appear as bands on the gel, and these bands can be viewed in the UV box after performing gel electrophoresis.
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American Society for Microbiology. (2019). Guidelines for biosafety in teaching laboratories. Retrieved from https://www.asm.org/getattachment/3c1eb38c-84d7-472f-aa9b-5d695985df21/ASM-Biosafety-Guidelines.pdf
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Bhatia, S., Tanveer, N. & Sardana, S. (2019). Introduction to pharmaceutical biotechnology. Bristol, United Kingdom: IOP publishing Limited
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Ndifon, W. & Dushoff, J. (2016). The Hayflick limit may determine the effective clonal diversity of naive T cells. The Journal of Immunology, 196(12), 4999-5004. https://doi.org/10.4049/jimmunol.1502343
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