WORLD OF MICROBIOLOGY AND IMMUNOLOGY VOL 2 - PART 4 doc

Protists WORLD OF MICROBIOLOGY AND IMMUNOLOGY 460 • • plex life cycles, as they usually live in more than one host in their lifetimes. The plant-like protists, or algae, are all photosynthetic autotrophs. These organisms form the base of many food chains. Other creatures depend on these protists either directly for food or indirectly for the oxygen they produce. Algae are responsible for over half of the oxygen produced by photo- synthesizing organisms. Many forms of algae look like plants, but they differ in many ways. Algae do not have roots, stems, or leaves. They do not have the waxy cuticle plants have to prevent water loss. As a result, algae must live in areas where water is readily available. Algae do not have multicellular gametangia as the plants do. They contain chlorophyll, but also contain other photosynthetic pigments. These pigments give the algae characteristic colors and are used to classify algae into various phyla. Other characteristics used to classify algae are energy reserve storage and cell wall composition. Members of the phylum Euglenophyta are known as euglenoids. These organisms are both autotrophic as well as heterotrophic. There are hundreds of species of euglenoids. Euglenoids are unicellular and share properties of both plants and animals. They are plant-like in that they contain chloro- phyll and are capable of photosynthesis. They do not have a cell wall of cellulose, as do plants; instead, they have a pelli- cle made of protein. Euglenoids are like animals in that they are motile and responsive to outside stimuli. One particular species, Euglena, has a structure called an eyespot. This area of red pigments is sensitive to light. An Euglena can respond to its environment by moving towards areas of bright light, where photosynthesis best occurs. In conditions where light is not available for photosynthesis, euglenoids can be het- erotrophic and ingest their food. Euglenoids store their energy as paramylon, a type of polysaccharide. Members of the phylum Bacillariophyta are called diatoms. Diatoms are unicellular organisms with silica shells. They are autotrophs and can live in marine or freshwater envi- ronments. They contain chlorophyll as well as pigments called carotenoids, which give them an orange-yellow color. Their shells resemble small boxes with lids. These shells are covered with grooves and pores, giving them a decorated appearance. Diatoms can be either radially or bilaterally symmetrical. Diatoms reproduce asexually in an unique manner. The two halves of the shell separate, each producing a new shell that fits inside the original half. Each new generation, therefore, produces offspring that are smaller than the parent. As each generation gets smaller and smaller, a lower limit is reached, approximately one quarter the original size. At this point, the diatom produces gametes that fuse with gametes from other diatoms to produce zygotes. The zygotes develop into full sized diatoms that can begin asexual reproduction once more. When diatoms die, their shells fall to the bottom of the ocean and form deposits called diatomaceous earth. These deposits can be collected and used as abrasives, or used as an additive to give certain paints their sparkle. Diatoms store their energy as oils or carbohydrates. The dinoflagellates are members of the phylum Dinoflagellata. These organisms are unicellular autotrophs. Their cell walls contain cellulose, creating thick, protective plates. These plates contain two grooves at right angles to each other, each groove containing one flagellum. When the two flagella beat together, they cause the organism to spin through the water. Most dinoflagellates are marine organisms, although some have been found in freshwater environments. Dinoflagellates contain chlorophyll as well as carotenoids and red pigments. They can be free-living, or live in symbiotic relationships with jellyfish or corals. Some of the free-living dinoflagellates are bioluminescent. Many dinoflagellates pro- duce strong toxins. One species in particular, Gonyaulax catanella, produces a lethal nerve toxin. These organisms sometimes reproduce in huge amounts in the summertime, causing a red tide. There are so many of these organisms pres- ent during a red tide that the ocean actually appears red. When this occurs, the toxins that are released reach such high con- centrations in the ocean that many fish are killed. Dinoflagellates store their energy as oils or polysaccharides. The phylum Rhodophyta consists of the red algae. All of the 4,000 species in this phylum are multicellular (with the exception of a few unicellular species) and live in marine envi- ronments. Red algae are typically found in tropical waters and sometimes along the coasts in cooler areas. They live attached to rocks by a structure called a holdfast. Their cell walls con- tain thick polysaccharides. Some species incorporate calcium carbonate from the ocean into their cell walls as well. Red algae contain chlorophyll as well as phycobilins, red and blue pigments involved in photosynthesis. The red pigment is called phycoerythrin and the blue pigment is called phyco- cyanin. Phycobilins absorb the green, violet, and blue light waves that can penetrate deep water. These pigments allow the red algae to photosynthesize in deep water with little light available. Reproduction in these organisms is a complex alter- nation between sexual and asexual phases. Red algae store their energy as floridean starch. The 1,500 species of brown algae are the members of the phylum Phaeophyta. The majority of the brown algae live in marine environments, on rocks in cool waters. They contain chlorophyll as well as a yellow-brown carotenoid called fucoxanthin. The largest of the brown algae are the kelp. The kelp use holdfasts to attach to rocks. The body of a kelp is called a thallus, which can grow as long as 180 ft (60 m). The thallus is composed of three sections, the holdfast, the stipe, and the blade. Some species of brown algae have an air blad- der to keep the thallus floating at the surface of the water, where more light is available for photosynthesis. Brown algae store their energy as laminarin, a carbohydrate. The phylum Chlorophyta is known as the green algae. This phylum is the most diverse of all the algae, with greater than 7,000 species. The green algae contain chlorophyll as their main pigment. Most live in fresh water, although some marine species exist. Their cell walls are composed of cellu- lose, which indicates the green algae may be the ancestors of modern plants. Green algae can be unicellular, colonial, or multicellular. An example of a unicellular green alga is Chlamydomonas. An example of a colonial algae is Volvox. A Volvox colony is a hollow sphere of thousands of individual cells. Each cell has a single flagellum that faces the exterior of the sphere. The individual cells beat their flagella in a coordi- womi_P 5/7/03 11:11 AM Page 460 Protists WORLD OF MICROBIOLOGY AND IMMUNOLOGY 461 • • nated fashion, allowing the colony to move. Daughter colonies form inside the sphere, growing until they reach a certain size and are released when the parent colony breaks open. Spirogyra and Ulva are both examples of multicellular green algae. Reproduction in the green algae can be both sexual and asexual. Green algae store their energy as starch. The fungus-like protists resemble the fungi during some part of their life cycle. These organisms exhibit properties of both fungi and protists. The slime molds and the water molds are members of this group. They all obtain energy by decom- posing organic materials, and as a result, are important for recycling nutrients. They can be brightly colored and live in cool, moist, dark habitats. The slime molds are classified as either plasmodial or cellular by their modes of reproduction. The plasmodial slime molds belong to the phylum Myxomycota, and the cellular slime molds belong to the phy- lum Acrasiomycota. The plasmodial slime molds form a structure called a plasmodium, a mass of cytoplasm that contains many nuclei but has no cell walls or membranes to separate individual cells. The plasmodium is the feeding stage of the slime mold. It moves much like an amoeba, slowly sneaking along decay- ing organic material. It moves at a rate of 1 in (2.5 cm) per hour, engulfing microorganisms. The reproductive structure of plasmodial slime molds occurs when the plasmodium forms a stalked structure during unfavorable conditions. This structure produces spores that can be released and travel large distances. The spores land and produce a zygote that grows into a new plasmodium. The cellular slime molds exist as individual cells during the feeding stage. These cells can move like an amoeba as well, engulfing food along the way. The feeding cells repro- duce asexually through cell division. When conditions become unfavorable, the cells come together to form a large mass of cells resembling a plasmodium. This mass of cells can move as one organism and looks much like a garden slug. The mass eventually develops into a stalked structure capable of sexual reproduction. The water molds and downy mildews belong to the phy- lum Oomycota. They grow on the surface of dead organisms or plants, decomposing the organic material and absorbing nutri- ents. Most live in water or in moist areas. Water molds grow as a mass of fuzzy white threads on dead material. The difference between these organisms and true fungi is the water molds form flagellated reproductive cells during their life cycles. Many protists can cause serious illness and disease. Malaria, for example, is caused by the protist Plasmodium. Plasmodia are sporozoans and are transferred from person to Diatoms, an example of protists. womi_P 5/7/03 11:11 AM Page 461 Protoplasts and spheroplasts WORLD OF MICROBIOLOGY AND IMMUNOLOGY 462 • • person through female Anopheles mosquitoes. People who suffer from malaria experience symptoms such as shivering, sweating, high fevers, and delirium. African sleeping sick- ness , also known as African trypanosomiasis, is caused by another sporozoan, Trypanosoma. Trypanosoma is transmitted through the African tsetse fly. This organism causes high fever and swollen lymph nodes. Eventually the protist makes its way into the victim’s brain, where it causes a feeling of uncon- trollable fatigue. Giardiasis is another example of a disease caused by a protist. This illness is caused by Giardia, a sporo- zoan carried by muskrats and beavers. Giardiasis is character- ized by fatigue, cramps, diarrhea, and weight loss. Amoebic dysentery occurs when a certain amoeba, Entamoeba histolyt- ica, infects the large intestine of humans. It is spread through infected food and water. This organism causes bleeding, diar- rhea, vomiting, and sometimes death. Members of the kingdom Protista can also be very ben- eficial to life on Earth. Many species of red algae are edible and are popular foods in certain parts of the world. Red algae are rich in vitamins and minerals. Carageenan, a polysaccha- ride extracted from red algae, is used as a thickening agent in ice cream and other foods. Giant kelp forests are rich ecosys- tems, providing food and shelter for many organisms. Trichonymphs are flagellates that live in the intestines of ter- mites. These protozoans break down cellulose in wood into carbohydrates the termites can digest. The kingdom Protista is a diverse group of organisms. Some protists are harmful, but many more are beneficial. These organisms form the foundation for food chains, produce the oxygen we breathe, and play an important role in nutrient recycling. Many protists are economically useful as well. As many more of these unique organisms are discovered, humans will certainly enjoy the new uses and benefits protists provide. See also Eukaryotes PROTOPLASTS AND SPHEROPLASTS Protoplasts and spheroplasts Protoplasts and spheroplasts are altered forms of bacteria or yeast, in which the principal shape-maintaining structure of the bacteria is weakened. Each bacterium forms a sphere, which is the shape that allows the bacterium to withstand the rigors, particularly osmotic, of the fluid in which it resides. The term protoplast refers to the spherical shape assumed by Gram-positive bacteria. Spheroplast refers to the spherical shape assumed by Gram-negative bacteria. The dif- ference is essentially the presence of a single membrane, in the case of the protoplast, and the two membranes (inner and outer) of the Gram-negative spheroplasts. It is also possible to generate a gram-negative protoplast by the removal of the outer membrane. Thus, in essence, protoplast refers to a bac- terial sphere that is bounded by a single membrane and spher- oplast refers to a sphere that is bounded by two membranes. Bacteria are induced to form protoplasts or spheroplasts typically by laboratory manipulation. However, formation of the structures can occur naturally. Such bacteria are referred to as L-forms. Examples of bacterial genera that can produce L- forms include Bacillus, Clostridium, Haemophilus, Pseudomonas, Staphylococcus, and Vibrio. The peptidoglycan is the main stress-bearing layer of the bacterial cell wall and the peptidoglycan also gives the bacterium its shape. In the laboratory, weakening the peptido- glycan network in the cell wall generates both protoplasts and spheroplasts. By exposing bacteria to an enzyme called lysozyme, the interconnecting strands of the two particular sugars that form the peptidoglycan can be cut. When this is done, the peptidoglycan loses the ability to serve as a mechanical means of support. The situation in yeast is slightly different, as other com- ponents of the yeast cell wall are degraded in order to form the protoplast. The process of creating protoplasts and spheroplasts must be done in a solution in which the ionic composition and concentration of the fluid outside of the bacteria is the same as that inside the bacteria. Once the structural support of the peptidoglycan is lost, the bacteria are unable to control their response to differences in the ionic composition between the bacterial interior and exterior. If the inner concentration is greater than the outer ionic concentration, water will flow into the bacterium in an attempt to achieve an ionic balance. The increased volume can be so severe that the bacteria will burst. Conversely, if the inner ionic concentration is less than the exterior, water will exit the bacterium, in an attempt to dilute the surroundings. The bacteria can shrivel to the point of death. Preservation of ionic balance is required to ensure that bacteria will not be killed during their transformation into either the protoplast or the spheroplast form. Living proto- plasts and spheroplasts are valuable research tools. The mem- brane balls that are the protoplasts or spheroplasts can be induced to fuse more easily with similar structures as well as with eukaryotic cells. This facilitates the transfer of genetic material between the two cells. As well, the sequential manu- facture of spheroplasts and protoplasts in Gram-negative bac- teria allows for the selective release of the contents of the periplasm. This approach has been popular in the identifica- tion of the components of the periplasm, and in the localiza- tion of proteins to one or the other of the Gram-negative membranes. For example, if a certain protein is present in a spheroplast population—but is absent from a protoplast popu- lation—then the protein is located within the outer membrane. See also Bacterial ultrastructure; Biotechnology; Trans- formation PROTOZOA Protozoa Protozoa are a very diverse group of single-celled organisms, with more than 50,000 different types represented. The vast majority are microscopic, many measuring less than 1/200 mm, but some, such as the freshwater Spirostomun, may reach 0.17 in (3 mm) in length, large enough to enable it to be seen with the naked eye. womi_P 5/7/03 11:11 AM Page 462 Protozoa WORLD OF MICROBIOLOGY AND IMMUNOLOGY 463 • • Scientists have discovered fossilized specimen of proto- zoa that measured 0.78 in (20 mm) in diameter. Whatever the size, however, protozoans are well-known for their diversity and the fact that they have evolved under so many different conditions. One of the basic requirements of all protozoans is the presence of water, but within this limitation, they may live in the sea, in rivers, lakes, stagnant ponds of freshwater, soil, and in some decaying matters. Many are solitary organisms, but some live in colonies; some are free-living, others are sessile; and some species are even parasites of plants and animals (including humans). Many protozoans form complex, exqui- site shapes and their beauty is often greatly overlooked on account of their diminutive size. The protozoan cell body is often bounded by a thin pli- able membrane, although some sessile forms may have a toughened outer layer formed of cellulose, or even distinct shells formed from a mixture of materials. All the processes of life take place within this cell wall. The inside of the mem- brane is filled with a fluid-like material called cytoplasm, in which a number of tiny organs float. The most important of these is the nucleus, which is essential for growth and repro- duction. Also present are one or more contractile vacuoles, which resemble air bubbles, whose job it is to maintain the correct water balance of the cytoplasm and also to assist with food assimilation. Protozoans living in salt water do not require contractile vacuoles as the concentration of salts in the cytoplasm is simi- lar to that of seawater and there is therefore no net loss or gain of fluids. Food vacuoles develop whenever food is ingested and shrink as digestion progresses. If too much water enters the cell, these vacuoles swell, move towards the edge of the cell wall and release the water through a tiny pore in the membrane. Some protozoans contain the green pigment chlorophyll more commonly associated with higher plants, and are able to manufacture their own foodstuffs in a similar manner to plants. Others feed by engulfing small particles of plant or ani- mal matter. To assist with capturing prey, many protozoans have developed an ability to move. Some, such as Euglena and Trypanosoma are equipped with a single whip like flagella which, when quickly moved back and forth, pushes the body through the surrounding water body. Other protozoans (e.g., Paramecium) have developed large numbers of tiny cilia around the membrane; the rhythmic beat of these hairlike structures propel the cell along and also carry food, such as bacteria, towards the gullet. Still others are capable of chang- ing the shape of their cell wall. The Amoeba, for example, is capable of detecting chemicals given off by potential food par- ticles such as diatoms, algae, bacteria or other protozoa. As the cell wall has no definite shape, the cytoplasm can extrude to form pseudopodia (Greek pseudes, “false”; pous, “foot”) in various sizes and at any point of the cell surface. As the Amoeba approaches its prey, two pseudopodia extend out from the main cell and encircle and engulf the food, which is then slowly digested. Various forms of reproduction have evolved in this group, one of the simplest involves a splitting of the cell in a process known as binary fission. In species like amoeba, this process takes place over a period of about one hour: the nucleus divides and the two sections drift apart to opposite ends of the cell. The cytoplasm also begins to divide and the cell changes shape to a dumb-bell appearance. Eventually the cell splits giving rise to two identical “daughter” cells that then resume moving and feeding. They, in turn, can divide further in this process known as asexual reproduction, where only one individual is involved. Some species that normally reproduce asexually, may occasionally reproduce through sexual means, which involves the joining, or fusion, of the nuclei from two different cells. In the case of paramecium, each individual has two nuclei: a larger macronucleus that is responsible for growth, and a much smaller micronucleus that controls reproduction. When paramecium reproduce by sexual means, two individuals join in the region of the oral groove—a shallow groove in the cell membrane that opens to the outside. When this has taken place, the macronuclei of each begins to disintegrate, while the micronucleus divides in four. Three of these then degener- ate and the remaining nucleus divides once again to produce two micronuclei that are genetically identical. The two cells then exchange one of these nuclei that, upon reaching the other individual’s micronucleus, fuse to form what is known as a zygote nucleus. Shortly afterwards, the two cells separate but within each cell a number of other cellular and cytoplas- mic divisions will continue to take place, eventually resulting in the production of four daughter cells from each individual. Protozoans have evolved to live under a great range of environmental conditions. When these conditions are unfavor- able, such as when food is scarce, most species are able to enter an inactive phase, where cells become non-motile and secrete a surrounding cyst that prevents desiccation and pro- tects the cell from extreme temperatures. The cysts may also serve as a useful means of dispersal, with cells being borne on the wind or on the feet of animals. Once the cyst reaches a more favorable situation, the outer wall breaks down and the cell resumes normal activity. Many species are of considerable interest to scientists, not least because of the medical problems that many cause. The tiny Plasmodium protozoan, the cause of malaria in humans, is responsible for hundreds of millions of cases of ill- ness each year, with many deaths occurring in poor countries. This parasite is transferred from a malarial patient to a healthy person by the bite of female mosquitoes of the genus Anopheles. As the mosquito feeds on a victim’s blood the par- asites pass from its salivary glands into the open wound. From there, they make their way to the liver where they multiply and later enter directly into red blood cells. Here they multiply even further, eventually causing the blood cell to burst and release from 6-36 infectious bodies into the blood plasma. A mosquito feeding on such a patient’s blood may absorb some of these organisms, allowing the parasite to complete its life cycle and begin the process all over again. The shock of the release of so many parasites into the human blood stream results in a series of chills and fevers—typical symptoms of malaria. Acute cases of malaria may continue for some days or even weeks, and may subside if the body is able to develop immunity to the disease. Relapses, however, are common and womi_P 5/7/03 11:11 AM Page 463 Prusiner, Stanley WORLD OF MICROBIOLOGY AND IMMUNOLOGY 464 • • malaria is still a major cause of death in the tropics. Although certain drugs have been developed to protect people from Plasmodium many forms of malaria have now developed, some of which are even immune to the strongest medicines. While malaria is one of the best known diseases known to be caused by protozoans, a wide range of other equally dev- astating ailments are also caused by protozoan infections. Amoebic dysentery, for example, is caused by Entamoeba his- tolytica.; African sleeping sickness, which is spread by the bite of the tsetse fly, is caused by the flagellate protozoan Trypanosoma; a related species T. cruzi causes Chagas’ dis- ease in South and Central America; Eimeria causes coccidio- sis in rabbits and poultry; and Babesia, spread by ticks, causes red water fever in cattle. Not all protozoans are parasites however, although this is by far a more specialized life style than that adopted by free- living forms. Several protozoans form a unique, nondestruc- tive, relationship with other species, such as those found in the intestine of wood-eating termites. Living in the termites’ intes- tines the protozoans are provided with free board and lodgings as they ingest the wood fibers for their own nutrition. In the process of doing so, they also release proteins which can be absorbed by the termite’s digestive system, which is otherwise unable to break down the tough cellulose walls of the wood fibers. Through this mutualistic relationship, the termites ben- efit from a nutritional source that they could otherwise not digest, while the protozoans receive a safe home and steady supply of food. See also Amoebic dysentery; Entamoeba histolytica; Epidemiology, tracking diseases with technology; Waste water treatment; Water quality PRUSINER, STANLEY (1942- ) Prusiner, Stanley American physician Stanley Prusiner performed seminal research in the field of neurogenetics, identifying the prion, a unique infectious pro- tein agent containing no DNA or RNA. Prusiner was born on in Des Moines, Iowa. His father, Lawrence, served in the United States Navy, moving the fam- ily briefly to Boston where Lawrence Prusiner enrolled in Naval officer training school before being sent to the South Pacific. During his father’s absence, the young Stanley lived with his mother in Cincinnati, Ohio. Shortly after the end of World War II, the family returned to Des Moines where Stanley attended primary school and where his brother, Paul, was born. In 1952, the family returned to Ohio where Lawrence Prusiner worked as a successful architect. In Ohio, Prusiner attended the Walnut Hills High School, before being accepted by the University of Pennsylvania where he majored in Chemistry. At the University, besides numerous science courses, he also had the opportunity to broaden his studies in subjects such as philoso- phy, the history of architecture, economics, and Russian his- tory. During the summer of 1963, between his junior and senior years, he began a research project on hypothermia with Sidnez Wolfson in the Department of Surgery. He worked on the project throughout his senior year and then decided to stay on at the University to train for medical school. During his second year of medicine, Prusiner decided to study the surface fluorescence of brown adipose tissue (fatty tissue) in Syrian golden hamsters as they arose from hibernation. This research allowed him to spend much of his fourth study year at the Wenner-Gren Institute in Stockholm working on the metabo- lism of isolated brown adipocytes. At this he began to seri- ously consider pursuing a career in biomedical research. Early in 1968, Prusiner returned to the U.S. to complete his medical studies. The previous spring, he had been given a position at the National Institutes of Health (NIH) on com- pleting an internship in medicine at the University of California San Francisco (UCSF). During that year, he met his wife, Sandy Turk, who was teaching mathematics to high school students. At the NIH, he worked on the glutaminase family of enzymes in Escherichia coli and as the end of his time at the NIH began to near, he examined the possibility of taking up a postdoctoral fellowships in neurobiology. Eventually, however, he decided that a residency in neurology was a better route to developing a rewarding career in research as it offered him direct contact with patients and therefore an opportunity to learn about both the normal and abnormal nerv- ous system. In July 1972, Prusiner began a residency at UCSF in the Department of Neurology. Two months later, he admit- ted a female patient who was exhibiting progressive loss of memory and difficulty performing some routine tasks. This was his first encounter with a Creutzfeldt-Jakob disease (CJD) patient and was the beginning of the work to which he has dedicated most of his life. In 1974, Prusiner accepted the offer of an assistant pro- fessor position from Robert Fishman, the Chair of Neurology at UCSF, and began to set up a laboratory to study scrapie, a parallel disease of human CJD found in sheep. Early on in this endeavor, he collaborated with William Hadlow and Carl Eklund at the Rocky Mountain Laboratory in Hamilton, Montana, from whom he learnt much about the techniques of handling the scrapie agent. Although the agent was first believed to be a virus, data from the very beginning suggested that this was a novel infectious agent, which contained no nucleic acid. It confirmed the conclusions of Tikvah Alper and J. S. Griffith who had originally proposed the idea of an infec- tious protein in the 1960s. The idea had been given little cre- dence at that time. At the beginning of his research into prion diseases, Prusiner’s work was fraught with technical difficul- ties and he had to stand up to the skepticism of his colleagues. Eventually he was informed by the Howard Hughes Medical Institute (HHMI) that they would not renew their financial support and by UCSF that he would not be promoted to tenure. The tenure decision was eventually reversed, however, enabling Prusiner to continue his work with financial support from other sources. As the data for the protein nature of the scrapie agent accumulated, Prusiner grew more confident that his findings were not artifacts and decided to summarize his work in a paper, published in 1982. There he introduced the term “prion,” derived from “proteinaceous” and ‘infectious” particle and challenged the scientific community to attempt to womi_P 5/7/03 11:11 AM Page 464 Pseudomonas WORLD OF MICROBIOLOGY AND IMMUNOLOGY 465 • • find an associated nucleic acid. Despite the strong convictions of many, none was ever found. In 1983, the protein of the prion was found in Prusiner’s laboratory and the following year, a portion of the amino acid sequence was determined by Leroy Hood. With that knowl- edge, molecular biological studies of prions ensued and an explosion of new information followed. Prusiner collaborated with Charles Weissmann on the molecular cloning of the gene encoding the prion protein (PrP). Work was also done on link- ing the PrP gene to the control of scrapie incubation times in mice and on the discovery that mutations within the protein itself caused different incubation times. Antibodies that pro- vided an extremely valuable tool for prion research were first raised in Prusiner’s lab and used in the discovery of the nor- mal form of PrP protein. By the early 1990s, the existence of prions as causative agents of diseases like CJD in humans and bovine spongiform encephalopathy (BSE) in cows, came to be accepted in many quarters of the scientific community. As pri- ons gained wider acceptance among scientists, Prusiner received many scientific prizes. In 1997, Prusiner was awarded the Nobel Prize for medicine. See also BSE and CJD disease; Infection and resistance; Viral genetics PSEUDOMEMBRANOUS COLITIS Pseudomembranous colitis Pseudomembranous colitis is severe inflammation of the colon in which raised, yellowish plaques, or pseudomembranes, develop on the mucosal lining. The plaques consist of clumps of dead epithelial cells from the colon, white blood cells, and fibrous protein. Pseudomembranous colitis is usually associated with antibiotic use. When the normal balance of the flora in the colon is disturbed, pathogenic strains of the bacillus Clostridium difficile may proliferate out of control and produce damaging amounts of cytotoxins known as cytotoxins A and B. C. difficile toxins often cause diarrhea and mild inflam- mation of the colon. Less frequently, the condition may progress further, causing ulceration and formation of the pseudomembranous plaques. Pseudomembranous colitis is most common in health care facilities such as hospitals and nursing homes, where an individual is most likely to be immune-compromised and to come into contact with persist- ent, heat-resistant C. difficile spores by the fecal-oral route. Thus, the best way to prevent it is meticulous cleanliness, cou- pled with avoiding the overuse of antibiotics. Mild symptoms such as diarrhea often disappear spon- taneously soon after the antibiotics are discontinued. Ironically, severe antibiotic-associated colitis must generally be treated with additional antibiotics to target the C. difficile pathogen. Benign intestinal flora such as lactobacillus or non- pathogenic yeast may be administered orally or rectally. Supportive therapies such as intravenous fluids are used as in other cases of ulcerative colitis. In rare cases, surgery to remove the damaged section of colon may be required. While antibiotic use is the most common precipitating cause of pseudomembranous colitis, occasionally the condi- tion may result from chemotherapy, bone marrow transplanta- tion, or other causes. See also Microbial flora of the stomach and gastroin- testinal tract P SEUDOMONAS Pseudomonas The genus Pseudomonas is made up of Gram-negative, rod- shaped bacteria that inhabit many niches. Pseudomonas species are common inhabitants of the soil, water, and vegeta- tion. The genus is particularly noteworthy because of the ten- dency of several species to cause infections in people who are already ill, or whose immune systems are not operating prop- erly. Such infections are termed opportunistic infections. Pseudomonas rarely causes infections in those whose immune systems are fully functional. The disease-causing members of the genus are therefore prevalent where illness abounds. Pseudomonas are one of the major causes of noso- comial (hospital acquired) infections. Bacteria in this genus not only cause infections in man, but also cause infections in plants and animals (e.g., horses). For example, Pseudomonas mallei causes ganders disease in horses. The species that comprise the genus Pseudomonas are part of the wider family of bacteria that are classified as Pseudomonadaceae. There are more than 140 species in the genus. The species that are associated with opportunistic infections include Pseudomonas aeruginosa, Pseudomonas maltophilia, Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas cepacia, Pseudomonas stutzeri, and Pseudomonas putrefaciens. Pseudomonas aeruginosa is prob- ably the most well-known member of the genus. Pseudomonas are hardy microorganisms, and can grow on almost any available surface where enough moisture and nutrients are present. Members of the genus are prone to form the adherent bacterial populations that are termed biofilms. Moreover, Pseudomonas aeruginosa specifically change their genetic behavior when on a surface, such that they produce much more of the glycocalyx material than they produce when floating in solution. The glycocalyx-enmeshed bacteria become extremely resistant to antibacterial agents and immune responses such as phagocytosis. In the hospital setting Pseudomonas aeruginosa can cause very serious infections in people who have cancer, cystic fibrosis, and burns. Other infections in numerous sites in the body, can be caused by Pseudomonas spp. Infections can be site-specific, such as in the urinary tract or the respiratory sys- tem. More widely disseminated infections (termed systemic infections) can occur, particularly in burn victims and those whose immune systems are immunosuppressed. For those afflicted with cystic fibrosis, the long-lasting lung infection caused by Pseudomonas aeruginosa can ulti- mately prove to be fatal. The bacteria have a surface that is altered from their counterparts growing in natural environ- womi_P 5/7/03 11:11 AM Page 465 Psychrophilic bacteria WORLD OF MICROBIOLOGY AND IMMUNOLOGY 466 • • ments. One such alteration is the production of a glycocalyx around the bacteria. The bacteria become very hard for the immune system to eradicate. The immune response eventually damages the epithelial cells of the lung. So much so, some- times, that lung function is severely compromised or ceases. Another bacterium, Pseudomonas cepacia, is also an opportunistic cause of lung infections in those afflicted with cystic fibrosis. This species is problematic because it is resist- ant to more antibiotics than is Pseudomonas aeruginosa. Glycocalyx production in some strains of Pseudomonas aeruginosa can be so prodigious that colonies growing on solid media appear slimy. Indeed, some species produce such mucoid colonies that the colonies will drip onto the lid of the agar plate when the plate is turned upside down. These slimy growths are described as mucoid colonies, and are often a hall- mark of a sample that has been recovered from an infection. Disease-causing species of Pseudomonas can possess a myriad of factors in addition to the glycocalyx that enable a bacterium to establish an infection. The appendages known as pili function in adherence to host cells. A component of the outer membrane possesses an endotoxin. Finally, a number of exotoxins and extracellular enzymes can cause damage at a distance from the bacterium. One such exotoxin, which is called toxin A, is extremely potent, and may be the prime cause of damage by the bacteria in infections. Some species, especially Pseudomonas aeruginosa are a problem in hospitals. By virtue of their function, hospitals are a place where many immunocompromised people are found. This is an ideal environment for an opportunistic dis- ease-causing bacterium. Moreover, Pseudomonas aeruginosa has acquired resistance to a number of commonly used antibi- otics. As yet, a vaccine to the bacterium does not exist. Prevention of the spread of Pseudomonas involves the obser- vance of proper hygiene, including handwashing. See also Bacteria and bacterial infection; Infection and resist- ance; Lipopolysaccharide and its constituents PSYCHROPHILIC BACTERIA Psychrophilic bacteria Psychrophilic (“cold loving”) microorganisms, particularly bacteria, have a preferential temperature for growth at less than 59° Fahrenheit (15° Celsius). Bacteria that can grow at such cold temperatures, but which prefer a high growth tem- perature, are known as psychrotrophs. The discovery of psychrophilic microorganisms and the increasing understanding of their functioning has increased the awareness of the diversity of microbial life on Earth. So far, more than 100 varieties of psychrophilic bacteria have been isolated from the deep sea. This environment is very cold and tends not to fluctuate in temperature. Psychrophilic bacte- ria are abundant in the near-freezing waters of the Arctic and the Antarctic. Indeed, in Antarctica, bacteria have been iso- lated from permanently ice-covered lakes. Other environ- ments where psychrophilic bacteria have been include high altitude cloud droplets. Psychrophilic bacteria are truly adapted for life at cold temperatures. The enzymes of the bacteria are structurally unstable and fail to operate properly even at room (or ambient) temperature. Furthermore, the membranes of psychrophilic bacteria contain much more of a certain kind of lipid than is found in other types of bacteria. The lipid tends to be more pli- able at lower temperature, much like margarine is more pliable than butter at refrigeration temperatures. The increased fluid- ity of the membrane makes possible the chemical reactions that would otherwise stop if the membrane were semi-frozen. Some psychrophiles, particularly those from the Antarctic, have been found to contain polyunsaturated fatty acids, which generally do not occur in prokaryotes. At room temperature, the membrane of such bacteria would be so fluid that the bac- terium would die. Aside from their ecological curiosity, psychrophilic bacteria have practical value. Harnessing the enzymes of these organisms allows functions such as the cleaning of clothes in cold water to be performed. Furthermore, in the Arctic and Antarctic ecosystems, the bacteria form an important part of the food chain that supports the lives of more complex crea- tures. In addition, some species of psychrophiles, including Listeria monocytogenes are capable of growth at refrigeration temperatures. Thus, spoilage of contaminated food can occur, which can lead to disease if the food is eaten. Listeriosis, a form of meningitis that occurs in humans, is a serious health threat, especially to those whose immune system is either not mature or is defective due to disease or therapeutic efforts. Other examples of such disease-causing bacteria include Aeromonas hydrophila, Clostridium botulinum, and Yersinia enterocolitica. See also Extremophiles PUBLIC HEALTH , CURRENT ISSUES Public health, current issues Public health is the establishment and maintenance of healthful living conditions for the general population. This goal requires organized effort from all levels of government. Underlying the current concerns in public health are three principle aims of public health efforts. First is the assessment and monitoring of populations, from the community level to the national level, to identify populations who are at risk for whatever health prob- lem is being considered. For example, public health efforts have shown that aboriginals in Canada are especially prone to developing diabetes. The second “plank” of public health is the formulation of policies to deal with the significant problems. Returning to the example, policies and strategies for action are now being formulated to reverse the trend. The third core pub- lic health function is to assure that everyone is able to receive adequate and affordable care and disease prevention services. There are many microbiological threats to public health. In order to maintain the three cores of public health, priorities must be established. In organizations such as the Centers for Disease Control and the World Health Organization, different divisions have been created to address the different concerns. Within each division the particular area of concern, such as womi_P 5/7/03 11:11 AM Page 466 Public health, current issues WORLD OF MICROBIOLOGY AND IMMUNOLOGY 467 • • food safety, can be simultaneously addressed at various levels, including basic research, policy development, and public awareness. In the aftermath of the September 11, 2001, terrorist attacks on targets in the United States, public perception of the health risks of what is commonly known as bioterrorism has been heightened. The ability to transport harmful microorgan- isms or their products, such as anthrax, through the mail or via dispersal in the air has made clear how vulnerable populations are to attack. Public health agencies have realized that the abil- ity to promptly respond to an incident is critical to any suc- cessful containment of the disease causing microbial threat. But the achievement of this response will require a huge effort from many public and private agencies, and will be extremely expensive. For example, it has been estimated that a response to each incident of bioterrorism, real or not, costs on the order of 50,000 dollars. Repeated mobilization of response teams would quickly sap the public health budget, at the cost of other programs. Thus, in the latter years of the twentieth century and the new century, the issue of bioterrorism and how to deal with it in a safe and economically prudent way has become a para- mount public health issue. Another public health issue that has become more important is the emergence of certain microbial diseases. In the emergence category, hemorrhagic diseases of viral origin, such as Ebola and Lassa fever are appearing more frequently. These diseases are terrifying due to their rapid devastation inflicted on the victim of infection, and because treatments are as yet rudimentary. The emergence of such diseases, which seems to be a consequence of man’s encroachment on envi- ronments that have been largely untouched until now, is a har- binger of things to come. Public health agencies are moving swiftly to understand the nature of these diseases and how to combat them. Diseases are also re-emerging. Tuberculosis is one example. Diseases such as tuberculosis were once thought to be a thing of the past, due to antibiotics and public health ini- tiatives. Yet, the numbers of people afflicted with such dis- eases is on the rise. One factor in the re-emergence of certain diseases is the re-acquisition of antibiotic resistance by bacte- ria . Another factor in the re-emergence of tuberculosis is the sharp increase in the number of immunocompromised indi- viduals that are highly susceptible to tuberculosis, such as those with acquired immune deficiency syndrome ( AIDS). The overuse and incomplete use of antibiotics has also enabled bacteria to develop resistance that can be passed on to subse- quent generations. Public health efforts and budgets are being Ciprofloxacin used to treat anthrax. womi_P 5/7/03 11:11 AM Page 467 Pyrex: construction, property, and uses in microbiology WORLD OF MICROBIOLOGY AND IMMUNOLOGY 468 • • re-directed to issues thought at one time to be dealt with and no longer a concern. Certain infectious diseases represent another increas- ingly important public health issue. Just a few decades ago AIDS was more of a curiosity, given its seeming confinement to groups of people who were often marginalized and ostra- cized. In the past decade, however, it has become clear that AIDS is an all-inclusive disease. Aside from the suffering that the illness inflicts, the costs of care for the increasingly debil- itated and dependent patients will constitute a huge drain on health care budgets in the decades to come. As a result, AIDS research to develop an effective vaccine or strategies that pro- long the vitality of those infected with the AIDS virus is a major public health issue and priority. Another public health issue of current importance is chronic bacterial and viral diseases. Conditions like fibromyalgia may have a bacterial or viral cause. The chronic and debilitating Lyme disease certainly has a bacterial cause. Moreover, the increasing use of surgical interventions to enhance the quality of life, with the installation of heart pace- makers, artificial joints, and the use of catheters to deliver and remove fluids from patients, has created conditions conducive for the explosion in the numbers of bacterial infections that result from the colonization of the artificial surfaces. Such bacterial biofilms have now been proven to be the source of infections that persist, sometimes without symptoms, in spite of the use of antibiotics. Such infections can be life threaten- ing, and their numbers are growing. As with the other current public health issues, chronic infections represent both a public health threat and a budget drain. A final area that has long been a public health concern is the safety of food and water. These have always been sus- ceptible to contamination by bacteria, protozoa and viruses, in particular. With the popularity of prepared foods, the monitor- ing of foods and their preparation has become both more urgent and more difficult for the limited number of inspectors to do. Water can easily become contaminated. The threat to water has become greater in the past twenty years, because of the increasing encroachment of civilization on natural areas, where the protozoan pathogens Giardia and Cryptosporidium normally live, and because of the appearance of more danger- ous bacterial pathogens, in particular Escherichia coli O157:H7. The latter organism is a problem in food as well. See also Bacteria and bacterial infection; Epidemics and pan- demics; Food safety; History of public health; Viruses and responses to viral infection PUBLIC HEALTH SYSTEMS • see HISTORY OF PUBLIC HEALTH PULSE-CHASE EXPERIMENT • see LABORATORY TECHNIQUES IN IMMUNOLOGY P YREX: CONSTRUCTION, PROPERTY, AND USES IN MICROBIOLOGY Pyrex: construction, property, and uses in microbiology Pyrex is a brand name of a type of glass that is constructed of borosilicate. The Corning Glass Company of Corning, New York, developed Pyrex. Chemically, as borosilicate implies, this type of glass is composed of silica and at least five percent (of the total weight of the elements in the glass) of a chemical called boric oxide. The combination and concentrations of these constituents confers great resistance to temperature change and corrosion by harsh chemicals, such as strong acids and alkalis, to whatever vessel is made of the borosilicate glass. This durability has made Pyrex glassware extremely useful in the microbiology laboratory. The development of Pyrex in 1924 by scientists at the Corning Company satisfied the demand for high quality scien- tific glassware that had began in the nineteenth century. Then, the glassware in existence was degraded by laboratory chemi- cals and became brittle when exposed to repeated cycles of heating and cooling. The formulation of Pyrex minimized the tendency of the material to expand and contract. This main- tained the accuracy of measuring instruments such as gradu- ated cylinders, and overcame the brittleness encountered upon repeated autoclave sterilization of the laboratory glassware. Pyrex glassware immediately found acceptance in the microbiology research community. The popularity of the glassware continues today, despite the development of heat and chemical resistant plastic polymers. Glass is still the pre- ferred container for growing bacteria. This is because the glass can be cleaned using harsh chemicals, which will completely remove any organic material that might otherwise adhere to the sides of the vessel. For applications where the chemical composition and concentrations of the medium components are crucial, such organic contaminants must be removed. Pyrex glassware is also used to manufacture graduated cylinders that are extremely accurate. In some applications, the exact volume of a liquid is important to achieve. This type of glassware is known as volumetric glassware. Plastic still can- not match the accuracy or the unchanging efficiency of volume delivery that is achieved by Pyrex volumetric glassware. Another application for borosilicate glass is in the meas- urement of optical density. For this application, typically spe- cially designed vials are filled with the solution or suspension of interest and then placed in the path of a beam of light in a machine known as a spectrophotometer. The amount of light that passes through the sample can be recorded and, with the inclusion of appropriate controls, can be used, for example, to determine the number of bacteria in the sample. Plastic mate- rial does not lend itself to optical density measurements, as the plastic can be cloudy. Thus, the vial itself would absorb some of the incoming light. Pyrex, however, can be made so as to be optically transparent. Growth flasks have even been made in which a so-called “side arm,” basically a test tube that is fused onto the flask, can be used to directly obtain optical density measurements without removing the culture from the flask. In the same vein, the use of optically transparent slabs of Pyrex as microscope slides is a fundamental tool in the micro- womi_P 5/7/03 11:11 AM Page 468 Pyrrophyta WORLD OF MICROBIOLOGY AND IMMUNOLOGY 469 • • biology laboratory. The heat resistance of the slide allows a specimen to be heated directly on the slide. This is important for stains such as the acid-fast stain for mycobacteria, in which heating of the samples is essential for the accurate staining of the bacteria. Also, as for the optical density measurements, the light microscopic examination of the bacterial sample depends upon the transparency of the support surface. Plastic is not an appropriate support material for slides. Another area in which Pyrex glassware is essential in a microbiology laboratory is in the pipelines required for the delivery of distilled water. Distillation of water is a process that requires the boiling of the water. The pipelines must be heat resistant. Also, because physical scrubbing of the pipelines is not feasible, the pipes must withstand the applica- tion of caustic chemicals to scour organic material off the inte- rior surface of the pipes. Other applications of borosilicate glassware in the microbiology laboratory include nondisposable Petri plates for the use of solid media, centrifuge tubes, titration cylinders, and the stopcocks that control the flow rate. Heat and chemically resistant plastics are widely used in the typical microbiology laboratory, particularly for routine, high-volume operations where cleaning and preparation of glassware for re-use is time-consuming and prone to error. However, the accuracy and advantages of Pyrex glassware ensure its continued use in the most modern of microbiology laboratories. See also Laboratory methods in microbiology; Microscopy PYRROPHYTA Pyrrophyta Approximately 2000 species of Pyrrophyta (from the Greek pyrrhos, meaning flames, and phyton, meaning plant) are known at present. Pyrrophyta have been identified in fossil deposits around the globe, from arctic to tropical seas, as well as in hypersaline waters, freshwater, and river deltas. Pyrrophyta are mostly unicellular microorganic Protists divided by botanists in two phyla, dinoflagellates and criptomonads. The taxonomic classification of Pyrrophyta is disputed by some zoologists who consider them members of the Protozoa kingdom. Cryptomonads for instance, are considered red-brownish algae of Cryptomonadida Order by botanists, and protozoans of Cryptophycea Class by zoologists. This controversy is due to the unusual characteristics of these two phyla, sharing features with both plants and animals. For instance, most species swim freely because of the spiraling Pyrex labware filled with colored liquid. womi_P 5/7/03 11:11 AM Page 469 [...]... source of radiation used in such treatments Iodine-131 and phosphorus- 32 are also mones Radioisotopes are also finding increasing use labeling, identification and study of immunological ce The study of microorganisms also relies heavily use of radioisotopes The identification of protein labeling of surface components of bacteria, and trac transcription and translation steps involved in nucle and protein... T-cell Leukemi (HTLV-1) The latter virus is a well-known tumor viru There are two groups of RNA tumor virus Oncovirinae and the Lentivirinae Examples of the firs include the Rous Sarcoma Virus, HTLV-1, and HTLV -2 is also known as hairy cell leukemia virus) A prominen ple of the second group is the Human Immunodeficienc (HIV) A characteristic of HIV and other members of the group is the long period of. .. against Q fever is not yet a standard A vaccine is available in Australia and parts of Europe, not yet been approved in North America Prevention of the transmission of the bacter humans involves the wearing of masks when around d • Q Q fever Mountain sheep, one of the natural hosts of the Q-fever bacterium Coxiella burnetii animals and the prompt disposal of placenta and other tissues resulting from... which contain iron in a nitrogen-containing heme gro final electron acceptor of the electron transfer chain is which produces water as a final product of cellular res The main function of the electron transfer cha synthesis of 32 molecules of ATP from the controlle tion of the eight molecules of NADH and two mole FADH2, made by the oxidation of one molecule of gl glycolysis and the citric acid cycle The... Bioremediation; Extremophiles RADIOISOTOPES AND THEIR USES IN MICROBIOLOGY AND IMMUNOLOGY Radioisotopes and their uses in microbiology and immunology Radioisotopes, containing unstable combinations of protons and neutrons, are created by neutron activation that involves the capture of a neutron by the nucleus of an atom Such a capture results in an excess of neutrons (neutron rich) Proton rich radioisotopes... period of weeks or months The rapid activation of the DNArepair pathway through p53 protein and the stress-inducible p21 protein as well as the extreme sensitivity of cells with genetic defects in DNA repair machinery support the view that the ability of the cell to repair irradiation-induced DNA damage is a limiting factor in deciding the extent of the mutagenic effects 47 8 • See also Evolution and evolutionary... from microorganisms Retroviruses are sphere-shaped viruses that c single strand or a couple of strands of RNA The shaped capsule of the virus consists of various prote capsule is studded on the outside with proteins called proteins In HIV, these receptor proteins bind to spe teins on T cells called CD4 receptors CD4 stands fo of differentiation, and CD type 4 is found on specific called helper cells... the management of the disease, and a promising non-invasive detection of incompatibility seems now possible by means of the polymerase chain reaction (PCR) analysis of cell-free fetal DNA circulating in the mother’s blood See also Antibody and antigen; Antibody formation and kinetics RHIZOBIUM-AGROBACTERIUM GROUP • see ECONOMIC USES AND BENEFITS OF MICROORGANISMS RHODOPHYTA Rhodophyta 48 8 • The red... into a series of nine se enzyme-catalyzed reactions, known as the citric aci These reactions are so named because the first reactio one molecule of citric acid (a 6-carbon molecule) f molecule of acetyl CoA (a 2- carbon molecule) and on cule of oxaloacetic acid (a 4- carbon molecule) A c round of the citric acid cycle expels two molecules o dioxide and regenerates one molecule of oxaloacetic a The citric... mechanism that involves transcription of one strand of the retroposon into RNA This RNA undergoes conformation change (looping) and provides a primer for the synthesis of single stranded cDNA The cDNA later serve as template for the synthesis of a double stranded DNA that is inserted in the genome by yet unknown mechanisms Transposons and retroposons seem to play a role in evolution and biology by promoting . domestic womi_Q 5/7/03 8:08 AM Page 47 1 Qualitative and quantitative analysis in microbiology WORLD OF MICROBIOLOGY AND IMMUNOLOGY 4 72 • • animals and the prompt disposal of placenta and other tissues resulting. one of the natural hosts of the Q-fever bacterium Coxiella burnetii. womi_Q 5/7/03 8:08 AM Page 4 72 Qualitative and quantitative analysis in microbiology WORLD OF MICROBIOLOGY AND IMMUNOLOGY 47 3 • • of. “proteinaceous” and ‘infectious” particle and challenged the scientific community to attempt to womi_P 5/7/03 11:11 AM Page 46 4 Pseudomonas WORLD OF MICROBIOLOGY AND IMMUNOLOGY 46 5 • • find an