(Environmental Factors /Lecture 9) 1-TEMPERATURE ADAPTATIONS Microbial cells are unable to control their temperature and therefore assume

the ambient temperature of their natural habitats. Their survival is dependent on adapting to whatever temperature variations are encountered in that habitat. The range of temperatures for microbial growth can be expressed as three cardinal temperatures. A- The minimum temperature is the lowest temperature that permits a microbes continued growth and metabolism; below this temperature, its activities are inhibited. B-The maximum temperature is the highest temperature at which growth and metabolism can proceed. If the temperature rises slightly above maximum, growth will stop, but if it continues to rise beyond that point, the enzymes and nucleic acids will eventually become permanently inactivated and the cell will die. This is why heat works so well as an agent in microbial control. C-The optimum temperature covers a small range, intermediate between the minimum and maximum, which promotes the fastest rate of growth and metabolism (rarely is the optimum a single point). a-A psychrophile is a microorganism that has an optimum temperature below 15C and is capable of growth at 0C. It is obligate with respect to cold and generally cannot grow above 20C. b-The majority of medically significant microorganisms are mesophiles, organisms that grow at intermediate temperatures. Although an individual species can grow at the extremes of 10C or 50C, the optimum growth temperatures (optima) of most mesophiles fall into the range of 2040C. Organisms in this group inhabit animals and plants as well as soil and water in temperate c-A thermophile is a microbe that grows optimally at temperatures greater than 45C. Such heat-loving microbes live in soil and water associated with volcanic activity and in habitats directly exposed to the sun. Thermophiles vary in heat requirements, with a general range of growth of 4580C. Most eucaryotic forms cannot survive above 60C, but a few thermophilic bacteria called hyperthermophiles, grow between 80C and 110C (currently thought to be the temperature limit endured by enzymes and cell structures). Strict thermophiles are so heat- tolerant that researchers may use an autoclave to isolate them in culture.

THE POPULATION GROWTH CURVE In reality, a population of bacteria does not maintain its potential growth rate and does not double endlessly, because in most systems numerous factors prevent the cells from continuously dividing at their maximum rate. Quantitative laboratory studies indicate that a population typically displays a predictable pattern, or growth curve, over time. The method traditionally used to observe the population growth pattern is a viable count technique, in which the total number of live cells is counted over a given time period. In brief, this method entails (1) placing a tiny number of cells into a sterile liquid medium; (2) incubating this culture over a period of several hours; (3) sampling the broth at regular intervals during incubation; (4) plating each sample onto solid media; and (5) counting the number of colonies present after incubation. Microbits 7.6 gives the details of this process. STAGES IN THE NORMAL GROWTH CURVE The system of batch culturing is closed, meaning that nutrients and space are finite and there is no mechanism for the removal of waste products. Data from an entire growth period of 3 to 4 days typically produce a curve with a series of phases termed the lag phase, the exponential growth (log) phase, the stationary phase, and the death phase. I-The lag phase is a relatively flat period on the graph when the population appears not to be growing or is growing at less than the exponential rate. Growth lags primarily because: (1) The newly inoculated cells require a period of adjustment, enlargement, and synthesis; (2) the cells are not yet multiplying at their maximum rate; (3) the population of cells is so sparse or dilute that the sampling misses them. II-the length of the lag period varies somewhat from one population to another. The cells reach the maximum rate of cell division during the exponential growth (log) phase, a period during which the curve increases geometrically. This phase will continue as long as cells have adequate nutrients and the environment is favorable. III-At the stationary growth phase, the population enters a survival mode in which cells stop growing or grow slowly. The curve levels off because the rate of cell inhibition or death balances out the rate of multiplication. IV- The decline in the growth rate is caused by depleted nutrients and oxygen, excretion of organic acids and other biochemical pollutants into the growth medium, and an increased density of cells. As the limiting factors intensify, cells begin to die in exponential numbers (literally perishing in their own wastes), and they are unable to multiply. The curve now dips downward as the death phase begins. The speed with which death occurs depends on the relative resistance of the species and how toxic the conditions are, but it is usually slower than the exponential growth phase. Viable cells often remain many weeks and months after this phase has begun. In the laboratory, refrigeration is used to slow the progression of the death phase so that cultures will remain viable as long as possible.

(Microbial Genetics /Lecture10) The basic unit of DNA structure is a nucleotide, a molecule composed of phosphate, deoxyribose sugar, and a nitrogenous base. The nucleotides covalently bond to form a sugar-phosphate linkage that becomes the backbone of each strand. Each sugar attaches in a repetitive pattern to two phosphates. One of the bonds is to the number 5_ (read five prime) carbon on deoxyribose, and the other is to the 3_ carbon, which confers a certain order and direction on each strand The nitrogenous bases, purines and pyrimidines, attach by covalent bonds at the 1_ position of the sugar. They span the center of the molecule and pair with appropriate complementary base from the other side of the helix. The paired bases are so aligned as to be joined by hydrogen bonds. Such weak bonds are easily broken, allowing the molecule to be unzipped into its complementary strands. Pairing of purines and pyrimidines is not random; it is dictated by the formation of hydrogen bonds between certain bases. Thus, in DNA, the purine adenine (A) pairs with the pyrimidine thymine (T), and the purine guanine (G) pairs with the pyrimidine cytosine (C).

(Transcription& Translation /Lecture 11)

Transcription
The first step in gene expression is the production of an RNA copy of the DNA sequence encoding the gene, a process called transcription. To understand the mechanism behind the transcription process, it is useful to focus first on RNA polymerase, the remarkable enzyme responsible for carrying it out Promoter Transcription starts at RNA polymerase binding sites called promoters on the DNA template strand. A promoter is a short sequence that is not itself transcribed by the polymerase that binds to it

.Lec 13/Antimicrobial chemotherapy....

DRUG, MICROBE, HOSTSOME BASIC INTERACTIONS (1) The drug is administered to the host via a designated route. Delivery is primarily by oral, circulatory, muscular, and cutaneous routes. (2) The drug is dissolved in body fluids. (3) The drug is delivered to the infected area (extracellular or intracellular). (4) The drug destroys the infectious agent or inhibits its growth. (5) The drug is eventually excreted or broken down by the hosts organs, ideally without harming them.

MECHANISMS OF DRUG ACTION Antimicrobial drugs function specifically in one of the following ways: (1) They inhibit cell wall synthesis; (2) they inhibit nucleic acid synthesis or function; (3) they inhibit protein synthesis; (4) they interfere with the function of the cell membrane.

OVERVIEW OF LABORATORY TECHNIQUES

The routes taken in specimen analysis are the following: (1) direct tests using microscopic, immunologic, or other specific methods that provide immediate clues as to the identity of the microbe or microbes in the sample and (2) cultivation, isolation, and identification of pathogens using a wide variety of general and specific tests . Most test results fall into two categories: presumptivedata, which place the isolated microbe (isolate) in a preliminary category such as a genus, and more specific, confirmatorydata, which provide more definitive evidence of a species Biochemical Testing The physiological reactions of bacteria to nutrients and other substrates provide excellent indirect evidence of the types of enzyme systems present in a particular species the polymerase chain reaction (PCR). This method can amplify DNA present in samples even in tiny amounts, which greatly improves the sensitivity of the test PCR tests are being used or developed for a wide variety of bacteria, viruses, protozoa, and fungi.

Microbiology

(IMMUNITYAND IMMUNIZATION /Lecture 16)

Active immunity occurs when an individual receives an immune stimulus (antigen) that activates the B and T cells, causing the body to produce immune substances such as antibodies. Active immunity is marked by several characteristics: (1) It is an essential attribute of an immunocompetent individual; (2) it creates a memory that renders the person ready for quick action upon reexposure to that same antigen; (3) it requires several days to develop; and (4) it lasts for a relatively long time, sometimes for life. Active immunity can be stimulated by natural or artificial means. Passive immunity occurs when an individual receives immune substances (antibodies) that were produced actively in the body of another human or animal donor. The recipient is protected for a time even though he or she has not had prior exposure to the antigen. It is characterized by: (1) lack of memory for the original antigen, (2) lack of production of new antibodies against that disease, (3) immediate onset of protection, and (4) short-term effectiveness, because antibodies have a limited period of function, and ultimately, the recipients body disposes of them. Passive immunity can also be natural or artificial in origin. Principles of Vaccine Preparation A vaccine must be considered from the standpoints of antigen selection, effectiveness, ease in administration, safety, and cost. In natural immunity, an infectious agent stimulates appropriate B and T lymphocytes and creates memory clones. In artificial active immunity, the objective is to obtain this same response with a modified version of the microbe or its components. A safe and effective vaccine should mimic the natural protective response, not cause a serious infection or other disease, have long-lasting effects in a few doses, and be easy to administer. Most vaccine preparations contain one of the following antigenic stimulants (1) killed whole cells or inactivated viruses, (2) live, attenuated cells or viruses, (3) antigenic components of cells or viruses, or (4) genetically engineered microbes or microbial antigens. Large, complex antigens such as whole cells or viruses are very effective immunogens. Depending on the vaccine, these are either killed or attenuated.