You are on page 1of 13
Cor i BACTERIAL CLASSIFICATION, STRUCTURE, AND REPLICATION he smallest bacteria (Chlamydia and Rickettsia) are just 0.1 to 0.2 um in diameter, whereas larger bacteria may bbe many microns in length. A newly described species is Ibundreds of times larger than the average bacterial cell and is visible to the naked eye. Most species, however, are approx- imately I jim in diameter and aze therefore visible with the use ofthe ight microscope, which has a resolution of 0.2 uz, In comparison, animal and plant cells are much larger, ranging from 7 im (the diameter of a red blood ell) several fect (the length of certain nerve cells. Differences between Eukaryotes and Prokaryotes Cll rom animals, plants, and fungi ae eukaryotes (Greek. for “true nucleus’), whereas bacteria, archae, and blue- green algae belong to the prokaryotes (Greck for “primitive ‘ucleus). The archae (archacbacteria) resemble bacteria in ‘most ways but represent a domain unique from bacteria and eukaryotes. In addition to lacking a nucleus and other organelles, the bacterial chromosome differs from the human chromosome in several ways. The chromosome ‘of a typical bacterium, such as Bscherichia coli, isa single, double-standed, cizcular molecule of deoxyribonucleic acid (DNA) containing approximately 5 million base pairs, an approximate length of 1.3 mm (ie, nearly 1000 times the diameter of the cell). The smallest bacterial chromosomes (Geom mycoplasmas) ate approximately one fourth of this size. In comparison, humans have two copies of 23 chromo: somes, which represent 2.9 10° base pairs 990 mm in length. Bacteria use a smaller ribosome, the 708 ribosome, and in most bacteria, a meshlike peptidoglycan cell wall surrounds the membranes to protect them against the envi ronment. Bacteria can survive, and in some cases grow, in hostile environments in which the osmotic pressure coaside the cell isso low that most eukaryotic cells would lyse, at temperature extremes (both hot and cold), with dryness, and with very dilute and diverse energy sources Bacteria have evolved their structures and functions to adapt lo these conditions. These and other distinguishing features are depicted in Figure 12-1 and outlined in Table 12 1, Several ofthese distinctions provide the basis for anti ricrobial action 106 Bacterial Classification Bacteria can be classified by their macroscopic and microscopic appearance, by characteristic growth and meta- bolic properties, by their antigenicity, and finally by their genotype. Macroscopic and Microscopie Distinetion ‘The initial distinction between bacteria can be made by. growth characteristics on different nutrient and selective ‘media. Bacteria grow in colonies; each colony is like a city of as many as a million or more organisms. The sum of theit characteristics provides the colony with distinguishing char- acteristics such as color, size, shape, and smell, The bacterias ability to resist certain antibioties, ferment specific sugars (eg, lactose, to distinguish Escherichia coli from Salmo. nella), to lyse erythrocytes (streptococcal hemolytic proper- ties), or to hydrolyze lipids (eg. clostridial lipase) can also be determined using the appropriate growth media, ‘The microscopic appearance, including size, shape, and configuration of the organisms (cocci, rods, curved, or spiral) and their ability to retain the Gram stain (gram-positive or gram-negative) are the primary means for distinguishing bacteria. A spherical bacterium such as Staphylococcus is a coccus, a rod-shaped bacterium such as E.coli is a bacillus, and the snakelike treponeme is a spirillum. In addition, Nocardia and Actinomyces species have branched filamentous appearances similar to those of fungi. Some bacteria form aggregates, such as the grapelike clusters of Staphylococcus ‘aureus or the diplococcus (two cells together) observed in Streptococcus or Neisseria species. Gram stains arapid, powerful, easy test that allows cini- cians to distinguish between the two major classes of bacte- ra, develop an initial diagnosis, and initiate therapy based on inherent differences in the bacteria (Figure 12-2). Bacte- ria are heat fixed or otherwise dried onto a slide, stained with crystal violet (Figure 12-3), a stain that is precipitated with iodine, and then the unbound and excess stain is removed by washing with the acetone-based decolorizer and water. A red counterstain, safranin, is added to stain any decolorized cells, This process (akes less than 10 minutes. For gram-positive bacteria, which turn purple, the stain gets trapped in a thick, cross-linked, meshlike struc ture, the peptidoglycan layer, which surrounds the cel. CHAPTER 12. BACTERIAL CLASSIFICATION, STRUCTURE, AND REPLICATION Prokaryote ol wall Single, supercoiled Peptaoaiyean | cular chromosome Flagelum Cytoplasm | plasmid Cell membrane fienim 708, (ste of celular foesomes respiration) Eukaryote Mitochondtion (sito of celular respiration) Cell membrane Nuclear membrane Lysosome ytopiasm ‘smooth ‘endoplasmic feticulum ough endoplasmic - relculum (rbosomes) 05 risesomes Golg apparatus FIGURE 12-1 Major features of prokaryotes and eukaryotes, 107 Gram-negative bacteria have a thin peptidoglycan layer that does not retain the crystal violet stain, so the cells ‘must be counterstained with safranin and turned red (Figure 12-4), A_ mnemonic device that may help is “p-PURPLE-POSITIVE” Gram staining loses dependability for bacteria that are starved (eg. old or stationary-phase cultures) or treated with antibiotics, owing to degradation of the peptoglycan. Bacte- ria that cannot be classified by Gram staining include myco- bacteria, which have a waxy outer shell and are distinguished with the acid-fast stain, and mycoplasmas, whieh have no peptidoglycan. Metabolic, Antigenic, and Genetic Distinction “The next level of classification ie based on the metabolic signature of the bacteria, including requirement for anaero- bic or aerobic environments, requirement for specific nutri- cents (eg, ability to ferment specific carbohydrates or use different compounds as a source of carbon for growth), and, production of characteristic metabolic products (acid, alcohols) and specific enzymes (eg.. staphylococcal cata: ase), Automated procedures for distinguishing enteric and ‘other bacteria have been developed; they analyze the growth, in different media and their microbial products and provide ‘a numerical biotype for each of the bacteria, ‘A particular strain of bacteria can be distinguished using antibodies to detect characteristic antigens on the bacteria (serotyping). These serologic tests can also be used to iden- fy organisms that are difficult (Treponema pallidum, the ‘organism responsible for syphilis) or too dangerous (eg. Francisella, the organism that causes tularemia), do not, {grow in the laboratory, ate associated with specific disease syndromes (eg., E. coli serotype O157:H7, responsible for hhemorthagic colitis), or need to be identified rapidly (eg. Fbrsti 121 lor onan o Etats and Parte Nor rps ‘gas, ung protaza, plants, animals Bacon 0 (approxima) 254m 053.0um Nuclear Structures Nuclous ‘lassie mambvane No ruclear membrane Ovommasomes ‘Sands of DNA dpi genome Sle, cular DNA haplold game Cytoplasmic Structures Netoshondia Present Absent ol bodies Present Absont Endoplasmic retouum Prosent Absent Rosomes(sodmontaton confor) 80 (60S + 408) 70S 608 + 305) Dyoplamie mamarane Contains ster Does nat conan terls (except mycoplasma) col wal Present fr fungi; otherise absent Poprosucton ‘Sowa and asoasal Movement ‘Corio aga, peasant Pespraton Via mitochondia Isa complex suture contain poten, ids, and pesidogheans ‘Aso binary sion) Sire tlaglum, If prosont Via ofoplasmic membrane edie tam Ht Sn Sts 4 Taubnan Mi ors: Corompory oral meri ad inmunay,S Lous, 1982, Nosy. 108 MEDICAL MICROBIOLOGY Lipoteichoic Cell wail Nutien-binetng 2 Protein Carter rote pepitiolycan FIGURE 12.2 Comparison of gram-positive and gram-negative ‘bacterial cell walls. A, gram-positive bacterium has a thick pep- Lidoglycan layer that contains teichoic and lipoteichoic acide. B, A gram-negative bacterium has a thin peptidoglycan layer and an ‘outer membrane that contains lipopolysaccharide, phospholipids, and proteins. The periplasmic space between the cytoplasmic and cuter membranes contsing transport, degradative, and cell walleya- Uhetic proteins, The outer membrane is joined to the cycoplasmic ‘membrane at adhesion points and i attached to the peptidoglycan by lipoprotein links. Streptococcus pyogenes, responsible for streptococcal pharyn- {itis). Serotyping is also used to subdivide bacteria below the species level for epidemiologic purposes. “The most precise method for classifying bacteria is by analysis oftheir genetic material, New methods distinguish bacteria by detection of specific characteristic DNA sequences. These techniques include DNA hybridization, polymerase chain reaction (PCR) amplification, DNA sequencing, and related techniques described in Chapter 5. ‘These genetic techniques do not require living or growing bacteria and can be used for rapid detection and identifica- tion of slow-growing organisms (e.g. mycobacteria, fungi) ‘or analysis of pathology samples of even very virulent bac- teria, The technology is now available for rapid analysis of the nudleie acid sequence of specific segments or the entire bacterial chromosome. The most common application ofthis technique is analysis of sequences of ribosomal DNA to detect the highly conserved sequences that identify a family Gram-Postive ‘Staphylococcus aureus sie cnt vet GB siep2 crmioine ip & op satan Bp Step 3 Decolonzer {alcohol er acetone) A bcos erphlosy ur oats @ / sortun Vo GI HORA Serechoe 8 FIGURE 123 Gram-stain morphology of bacteria, A, The crystal ‘violet of Gram stain is precipitated by Gram iodine and ie tapped fm the thick peptidoglycan layer in gram-positive bacteria. ‘he ecolorizer disperses the gram-negative outer membrane and washes the crystal violet from the thin layer of peptidoglycan fam-negative bacteria are visualized by the red counterstain. B, Bacterial morphologic. von eo Pecos (caps) er eo | cpa, we opopasmie os > Regen pone Surace prc chomasome Fagen) (Fact) Grau postive — GRAMNEGATIVE FIGURE 12-4 Gram-postive and gram-negative bacteria, A gram- positive bacterium has a thick layer of peptidoglycan (fling the purple space) (ef). A gram-negative bacterium bas a thin peptido- lycan layer (single black line) and an outer membrane (right) Structures in parentheses aze not found in all bacteria. Upon cell ‘vision, the membrane and peptidoglycan grow toward each other to form a division septum to separate the daughter cll