Mycobacterium tuberculosis Characteristics Mycobacterium tuberculosis is a rod-shaped (bacillus) bacterium that causes the disease tuberculosis in humans, as well

as other primates, hamsters, dogs, and guinea pigs (Figure 1). Since the organism is non-motile, it travels through the air on particles called droplet nuclei. Droplet nuclei, which range in size from 1 to 5 m, are introduced into the air when an infected person sneezes, coughs, etc. Normal air currents keep the particles airborne so that they can spread throughout an area. (See the Pathogenesis section for information on how an infection progresses.) In addition to being non-motile, M. tuberculosis is an obligate aerobe, meaning that the bacterium can only survive in an environment that contains oxygen. (See the Pathogenesis section to learn more about the environment in which M. tuberculosis lives.)

Figure 1. Scanning electron micrograph of Mycobacterium tuberculosis bacilli. Although Mycobacterium tuberculosis possesses a Gram-positive type cell wall, a cell wall with extensive peptidoglycan and no outer membrane, the bacterium does not stain with Gram stain reagents. Gram stain reagents are unable to penetrate the cell wall of the bacillus because layers of lipids surround the peptidoglycan in mycobacterium. Unlike most Gram-negative bacteria, which have a 5-20% lipid content by weight, M. tuberculosis and other mycobacterium are composed of up to 60% lipids. Many of these lipids are in the form of mycolic acids. (See the Adaptations section for more on the importance of high lipid content.) Since the Gram stain method proves ineffective on Mycobacterium tuberculosis, acid-fast staining must be used to make the bacilli visible under a microscope. In the Ziehl-Neelsen procedure, bacteria from a sputum (mucus coughed up from the lungs) sample are flooded with a basic solution of carbolfuchsin, a magenta dye. After heating the slide in a flame, the sample is washed with water and treated with acid-alcohol to decolorize the bacteria. Then, a counter stain of methylene blue is applied to the sample. When the process is complete, M. tuberculosis bacilli

appear pink because they retain the carbolfuchsin during the acid-alcohol decolorization step (Figure 2). Thus, mycobacterium is classified as acid-fast bacilli. Bacteria that are not acid-fast appear blue after the procedure.

Figure 2. Mycobacterium tuberculosis bacilli stained using the Ziehl-Neelsen method. Mycobacterium tuberculosis Pathogenesis Infection with Mycobacterium tuberculosis begins when droplet nuclei are inhaled into the upper respiratory tract through the mouth or the nose (Figure 2, box 1). From the upper respiratory tract, the bacilli travel through the bronchi until they reach the alveoli of the lungs (Figure 2, box 2). Once inside the lungs, alveolar macrophages (immune cells that engulf foreign particles) ingest the pathogenic organisms, but the mycobacterium does not die. Instead, the bacilli multiply within the macrophage hosts, causing the macrophages to rupture. The continued division of M. tuberculosis every 18 to 24 hours attracts more and more immune cells to the area. In an attempt to control the infection, some of these cells produce toxic substances that are supposed to kill the bacilli. The bacilli do not immediately die, however, so the release of toxic substances also damages the surrounding lung tissue. When macrophages and other cells of the immune system encircle this area of dead tissue, the lesion is called a tubercle or granuloma (Figure 1).

Figure 1. Mycobacterium tuberculosis bacilli visible within granuloma. This tissue sample was taken from the endometrial layer of the uterus. This is an example of tuberculosis disease persisting in an area of the body outside of the lungs. The interior of a tubercle consists of a gelatinous mass of host cells and bacilli that gives the damaged tissue a cheese-like consistency. Therefore, this type of tissue death is referred to as cessation necrosis. If the immune system is successful in preventing the M. tuberculosis bacilli from multiplying further, the caseous tubercles become walled-off and calcified (Figure 2, box 4). Although calcified lesions still contain viable bacteria, the bacteria cannot be spread to other individuals. When Mycobacterium tuberculosis lies dormant in the lungs, a person is said to have a latent tuberculosis infection. Such persons, who represent 8595% of infected individuals, show no overt symptoms of disease. In 5-15% of infected individuals, the immune system fails to prevent the infection from progressing and the interiors of the caseous lesions become liquefied. Liquefaction allows viable M. tuberculosis bacilli to spill out of the tubercles, leaving behind a cavity in the lungs (Figure 2, box 5). When these bacilli infect lower portions of the lungs or enter the bronchi the result is an active case of pulmonary tuberculosis disease. People with pulmonary tuberculosis are capable of spreading the disease to others through the bacteria in their sputum. They also manifest symptoms such as weight loss, weakness, night sweats, chest pain, and coughing up blood. If viable bacilli enter the bloodstream, M. tuberculosis can travel to organs of the body outside of the lungs (Figure 1, above and Figure 2, box 3). Known as extra-pulmonary tuberculosis, this form of the disease is rarely contagious.

Figure 2. Diagram depicting the pathogenesis of infection with Mycobacterium tuberculosis. As mentioned earlier, people with latent tuberculosis infection retain viable M. tuberculosis bacilli within their lungs, even though they are asymptomatic and not infectious. When a person with latent TB becomes immunosuppressed because of old age, lifestyle choices, illness, or a medical condition such as HIV, the dormant bacteria can reactivate within the calcified tubercles. Thus, people with latent tuberculosis infection always run the risk of developing active TB disease. (Source: http://bioweb.uwlax.edu/bio203/s2007/millard_ashl/)