Assignment in Medical Microbiology Lecture Pauline Ambat Arizaldo Castro Mary Joyce Menil Karizza Mae Samoranos HUB42

1. Describe the lines of proof Robert Koch used to definitively associate the bacterium Mycobacterium tuberculosis with the disease tuberculosis. How would this proof have been flawed if any of the tools he developed for studying bacterial diseases had not been available for his study of tuberculosis? (5 pts) Robert Koch studied the microorganisms responsible for anthrax, plague, cholera, malaria, sleeping sickness and several animal diseases. His name is usually associated with the development of staining and culture methods for bacteria and for the isolation of the slow growing etiological agent of tuberculosis Mycobacterium tuberculosis. This work had led to the formulation of Kochs postulates, which set out the conditions that must be met to confirm that a given bacterium is the cause of a particular disease. Later on, Koch examined patients and found the bacterium as blue rods against brown tissue. He cultured the tuberculosis cells on coagulated blood serum isolating pure Mycobacterium tuberculosis cultures that he then injected to guinea pigs which eventually acquired the disease. He successfully re-isolated Mycobacterium tuberculosis from the guinea pig thus, satisfying all of his postulates. Basically, his postulates were proven and he concluded that M. tuberculosis is the causative agent of tuberculosis. Through this, Koch made two important discoveries. First is that he discovered a way of staining human tissue for microscopic investigation showing Mycobacterium tuberculosis as thin blue rods on a brown background of human cells. Second is that he found out that M. tuberculosis is a slow growing, highly fastidious bacterium and would mature on coagulated serum. Koch began by examining tuberculosis patients for the presence of M. tuberculosis. If some of the techniques he used to study bacterial diseases were not available during his time like bacterial staining and cell culturing techniques, his evidences would have been flawed. If staining was not yet developed during his time, he would most probably not be able to visualize the rods from his collected diseased tissue. Without such visualization, he would not be able to have any organism to point out as the causative agent of tuberculosis. Also, his evidences would have been faulty if the organism could not be cultured outside the host and no then existing techniques will help him carry out such culturing. This means that the organism cannot grow without infecting humans. Since one of his premises is the transfer of an identified pathogenic microorganism from one host to another to cause the same disease, culturing and isolation methods were indeed pivotal to his work. In addition, if the organism is inoculated to other host and it doesnt produce the same effect as the original host had then there might be a problem. Last predicament that he might have encountered is the presence of another organism causing the same disease. His discussion and proofs would have been defective if another organism was detected aside from M. tuberculosis which can cause tuberculosis. An example is M. bovis which can also produce tuberculosis.

2. Imagine that all microorganisms suddenly disappeared from earth, why do you think that animals would eventually disappear? What will happen to plants? If by contrast, all higher organisms suddenly disappeared, what would happen to microorganisms? (10 pts) Microorganisms play a key role in the preservation of life on Earth. If by any chance they will disappear suddenly from this planet, animals will eventually disappear because of several reasons. In ecology, microbes play an important function as decomposers of dead organisms and this includes fungi and bacteria. Microorganisms are part of biological organization particularly the biotic component. They obtain energy from their environment that helps in the general flow of energy in communities and in most ecosystems. They release nutrients into the environment that are needed by other organisms. Microorganisms do this by eating dead organic matter. Microbial decomposers process the release of inorganic nutrients as they break down organic compounds during digestion. Simultaneous to this breakdown process is the use of the nutrients extracted by microbial decomposers for their own growth. In fact, there is a succession of different microflora that will act on dead organisms for their breakdown. If microorganisms will disappear, dead animals will not be decomposed and this will compromise energy flow hindering the processes that other dependent organisms must undertake. Without them, the world would quickly overrun with discarded food scraps, raw sewage and dead organisms. The loss of these microorganisms will also compensate the health of organisms that have a mutual relationship with a number of their representatives. An example is ruminants whose digestion is being aided by microorganisms. Another is human beings with harmless microorganisms found in their digestive tracts aiding in their digestion. Plants will also disappear because of lack of nutrients from dead organisms that will be returned by these microbial decomposers to the cycle of life. In contrast, if higher organisms will disappear, microorganisms will also be affected because of higher forms of organisms serve as their food source and habitats. With the absence of higher forms of organisms such as animals, the survival of microorganisms is also put at risk. Along with the disappearance of microorganisms, plants are in danger of extinction as well. Plants rely on Nitrogen for their health and growth. They only depend on soil for its supply because they cannot get Nitrogen directly from the environment. Through the process of fixation, Nitrogen from the atmosphere becomes available to the plants and this process takes place in mediation of microorganisms. Certain microbes, for instance Rhizobium and Cyanobacteria can convert the atmospheric Nitrogen into Nitrates and Nitrites, which are part of their metabolism and make it available for the plants. Also, plants will die because microorganisms are responsible for building fertile soil for plants to grow in. If microorganisms will disappear, no soil will be made for the terrestrial plants to live in. Meanwhile, if all higher organisms suddenly disappeared microorganisms cannot survive also. In order to survive and grow, microorganisms require a source of energy and nourishment. They need to be in environments that have optimum temperature, moisture, pH, and Oxygen, or other gases like Carbon dioxide. As there are billions of microorganisms on our planet, it can be said that different species need a wide range of things to survive, including decomposing dead matter into food. However, it is possible that some microorganisms like those belonging to Domain Archaeabacteria will be able to survive without the presence of higher forms of organisms such as

animals and plants. Since this group thrives in extreme conditions that are normally fatal to animals and plants, a sudden disappearance of plants and animals might not affect them at all. Overall, if one of these components becomes extinct there would be an imbalance in the biological organization. The consequences would be animals that rely on that particular part of the food chain will be unable to adapt to new or different food sources and will eventually die. Therefore, every life form has a place under the sun, because every living thing is important to someone or something else. 3. Prove that HIV is the causal agent for AIDS. (5 pts) Human Immunodeficiency Virus (HIV) from the genus Lentivirus is a retrovirus that contains identical strands of RNA, reverse transcriptase, and a phospholipid envelope with spikes termed as gp120 (glycoproteins with molecular weights of 120,000). The structural components of the virus render it the ability to produce Acquired Immunodeficiency Syndrome (AIDS), a condition manifested by infections of the fungus Candida albicans in different body areas, cytomegalovirus eye infections, tuberculosis, toxoplasmosis of the brain, Kaposis sarcoma and other rare infections of the immunocompromised. Infection begins when gp120 spikes embedded in the phospholipid envelope of the virus capacitates the virus to attach to CD4 receptors on the host cell (cells with these type of receptors include helper T cells, macrophages, and dendritic cells which are basically immune cells). Supplementary requirement for HIV infection are chemokine receptors such as CCR5 and CXCR4 (CCR5 is significant for the infection of macrophages while CXCR4 is involved mainly in the infection of T cells). Following viral attachment is host cell entry in which viral RNA is released and transcribed by reverse transcriptase into DNA that integrates to the host cells chromosomal DNA. This integrated DNA may or may not produce new HIV. In the process of being hidden inside the host cell, HIV becomes classified as a provirus which is not detected by the immune system. HIV-infected cells, instead of being killed, persist and serve as reservoir of latent HIV for years. This ability of HIV to become a provirus or latent is the basic reason why the bodys anti-HIV antibodies fail to hinder the progress of the infection. In addition other evasive mechanisms of the virus include cell-cell fusion and rapid antigenic changes. Everyday, 1 trillion HIVs are generated with short half-life of about 6 hours. These viruses must be cleared by the bodys defenses such as cytotoxic T cells, antibodies, and macrophages. However, there is a daily net loss of at least 20 million CD4 T cells due to their shortened life and the bodys inability to compensate by producing replacements for T cells. To sum it up, HIV destroys the immune system rendering the body inefficient to ward off opportunistic and normally weak pathogens thereby allowing them to establish themselves and cause infection to the body. Since this virus targets the immune cells, it incapacitates the bodys defenses by lessening their population and promoting the establishment of pathogens that will not be normally successful in establishing themselves in immunocompetent people. With such virulence, given the appropriate time, HIV will lead to the development of the complex of disorders that AIDS encompasses.

References:

(1) Smith RL, Smith TM. 2003. Elements of Ecology. 5th ed. Singapore; Pearson Education South Asia. pp 123-138 (2) Taylor GM, Stewart GR, Cooke M, Chaplin S, Ladva S, Kirup J, Palmer S, Yound DB. 2003. Koch's Bacillus a look at the first isolate of Mycobacterium tuberculosis from a modern perspective. [Internet] [cited 1 July 2012]. Available from Journal of Clinical Microbiology (3) Tortora GJ, Funke BR, Case CL. 2004. Microbiology An Introduction. 8th ed. Singapore; Pearson Education South Asia. pp. 545-553