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- 9.1: How Microbes Grow
- The bacterial cell cycle involves the formation of new cells through the replication of DNA and partitioning of cellular components into two daughter cells. In prokaryotes, reproduction is always asexual, although extensive genetic recombination in the form of horizontal gene transfer takes place, as will be explored in a different chapter. Most bacteria have a single circular chromosome; however, some exceptions exist.
- 9.2: Oxygen Requirements for Microbial Growth
- Ask most people “What are the major requirements for life?” and the answers are likely to include water and oxygen. Few would argue about the need for water, but what about oxygen? Can there be life without oxygen? The answer is that molecular oxygen is not always needed. The earliest signs of life are dated to a period when conditions on earth were highly reducing and free oxygen gas was essentially nonexistent.
- 9.3: The Effects of pH on Microbial Growth
- Bacteria are generally neutrophiles. They grow best at neutral pH close to 7.0. Acidophiles grow optimally at a pH near 3.0. Alkaliphiles are organisms that grow optimally between a pH of 8 and 10.5. Extreme acidophiles and alkaliphiles grow slowly or not at all near neutral pH. Microorganisms grow best at their optimum growth pH. Growth occurs slowly or not at all below the minimum growth pH and above the maximum growth pH.
- 9.4: Temperature and Microbial Growth
- Microorganisms thrive at a wide range of temperatures; they have colonized different natural environments and have adapted to extreme temperatures. Both extreme cold and hot temperatures require evolutionary adjustments to macromolecules and biological processes. Psychrophiles grow best in the temperature range of 0–15 °C whereas psychrotrophs thrive between 4°C and 25 °C. Mesophiles grow best at moderate temperatures in the range of 20 °C to about 45 °C. Pathogens are usually mesophiles.
- 9.5: Other Environmental Conditions that Affect Growth
- Microorganisms interact with their environment along more dimensions than pH, temperature, and free oxygen levels, although these factors require significant adaptations. We also find microorganisms adapted to varying levels of salinity, barometric pressure, humidity, and light.
- 9.6: Media Used for Bacterial Growth
- The study of microorganisms is greatly facilitated if we are able to culture them, that is, to keep reproducing populations alive under laboratory conditions. Culturing many microorganisms is challenging because of highly specific nutritional and environmental requirements and the diversity of these requirements among different species.
Thumbnail: Heavy rains cause runoff of fertilizers into Lake Erie, triggering extensive algal blooms, which can be observed along the shoreline. (credit: NASA)
Chapter 9: Microbial Growth
Next, the replication of DNA starts at a location on the circular chromosome called the origin of replication, where the chromosome is attached to the inner cell membrane.
Replication continues in opposite directions along the chromosome until the terminus is reached.
The center of the enlarged cell constricts until two daughter cells are formed, each offspring receiving a complete copy of the parental genome and a division of the cytoplasm (cytokinesis).
This process of cytokinesis and cell division is directed by a protein called FtsZ. FtsZ assembles into a Z ring on the cytoplasmic membrane. The Z ring is anchored by FtsZ-binding proteins and defines the division plane between the two daughter cells.
Additional proteins required for cell division are added to the Z ring to form a structure called the divisome.
The divisome activates to produce a peptidoglycan cell wall and build a septum that divides the two daughter cells.
Organisms categorized as mesophiles (“middle loving”) are adapted to moderate temperatures, with optimal growth temperatures ranging from room temperature (about 20 °C) to about 45 °C. As would be expected from the core temperature of the human body, 37 °C (98.6 °F), normal human microbiota and pathogens (e.g., E. coli, Salmonella spp., and Lactobacillus spp.) are mesophiles. The mesophiles occupy the same environments that humans do, in terms of foods that we eat, surfaces that we touch, and water that we swim in and drink.
9.4 Temperature and Microbial Growth
When the exploration of Lake Whillans started in Antarctica, researchers did not expect to find much life. Constant subzero temperatures and lack of obvious sources of nutrients did not seem to be conditions that would support a thriving ecosystem. To their surprise, the samples retrieved from the lake showed abundant microbial life. In a different but equally harsh setting, bacteria grow at the bottom of the ocean in sea vents (Figure 9.28), where temperatures can reach 340 °C (700 °F).
Microbes can be roughly classified according to the range of temperature at which they can grow. The growth rates are the highest at the optimum growth temperature for the organism. The lowest temperature at which the organism can survive and replicate is its minimum growth temperature . The highest temperature at which growth can occur is its maximum growth temperature . The following ranges of permissive growth temperatures are approximate only and can vary according to other environmental factors.
Organisms categorized as mesophile s (“middle loving”) are adapted to moderate temperatures, with optimal growth temperatures ranging from room temperature (about 20 °C) to about 45 °C. As would be expected from the core temperature of the human body, 37 °C (98.6 °F), normal human microbiota and pathogens (e.g., E. coli, Salmonella spp., and Lactobacillus spp.) are mesophiles.
Organisms called psychrotrophs , also known as psychrotolerant, prefer cooler environments, from a high temperature of 25 °C to refrigeration temperature about 4 °C. They are found in many natural environments in temperate climates. They are also responsible for the spoilage of refrigerated food.
The presence of Listeria in Jeni’s blood suggests that her symptoms are due to listeriosis , an infection caused by L. monocytogenes. Listeriosis is a serious infection with a 20% mortality rate and is a particular risk to Jeni’s fetus. A sample from the amniotic fluid cultured for the presence of Listeria gave negative results. Because the absence of organisms does not rule out the possibility of infection, a molecular test based on the nucleic acid amplification of the 16S ribosomal RNA of Listeria was performed to confirm that no bacteria crossed the placenta. Fortunately, the results from the molecular test were also negative.
Jeni was admitted to the hospital for treatment and recovery. She received a high dose of two antibiotics intravenously for 2 weeks. The preferred drugs for the treatment of listeriosis are ampicillin or penicillin G with an aminoglycoside antibiotic. Resistance to common antibiotics is still rare in Listeria and antibiotic treatment is usually successful. She was released to home care after a week and fully recovered from her infection.
L. monocytogenes is a gram-positive short rod found in soil, water, and food. It is classified as a psychrophile and is halotolerant. Its ability to multiply at refrigeration temperatures (4–10 °C) and its tolerance for high concentrations of salt (up to 10% sodium chloride [NaCl]) make it a frequent source of food poisoning. Because Listeria can infect animals, it often contaminates food such as meat, fish, or dairy products. Contamination of commercial foods can often be traced to persistent biofilms that form on manufacturing equipment that is not sufficiently cleaned.
Listeria infection is relatively common among pregnant women because the elevated levels of progesterone downregulate the immune system, making them more vulnerable to infection. The pathogen can cross the placenta and infect the fetus, often resulting in miscarriage, stillbirth, or fatal neonatal infection. Pregnant women are thus advised to avoid consumption of soft cheeses, refrigerated cold cuts, smoked seafood, and unpasteurized dairy products. Because Listeria bacteria can easily be confused with diphtheroids, another common group of gram-positive rods, it is important to alert the laboratory when listeriosis is suspected.
Go back to the previous Clinical Focus box.
The organisms retrieved from arctic lakes such as Lake Whillans are considered extreme psychrophile s (cold loving). Psychrophiles are microorganisms that can grow at 0 °C and below, have an optimum growth temperature close to
15 °C, and usually do not survive at temperatures above 20 °C. They are found in permanently cold environments such as the deep waters of the oceans. Because they are active at low temperature, psychrophiles and psychrotrophs are important decomposers in cold climates.
Organisms that grow at optimum temperatures of 50 °C to a maximum of 80 °C are called thermophiles (“heat loving”). They do not multiply at room temperature. Thermophiles are widely distributed in hot springs, geothermal soils, and manmade environments such as garden compost piles where the microbes break down kitchen scraps and vegetal material. Examples of thermophiles include Thermus aquaticus and Geobacillus spp. Higher up on the extreme temperature scale we find the hyperthermophiles , which are characterized by growth ranges from 80 °C to a maximum of 110 °C, with some extreme examples that survive temperatures above 121 °C, the average temperature of an autoclave. The hydrothermal vents at the bottom of the ocean are a prime example of extreme environments, with temperatures reaching an estimated 340 °C (Figure 9.28). Microbes isolated from the vents achieve optimal growth at temperatures higher than 100 °C. Noteworthy examples are Pyrobolus and Pyrodictium , archaea that grow at 105 °C and survive autoclaving. Figure 9.29 shows the typical skewed curves of temperature-dependent growth for the categories of microorganisms we have discussed.
Life in extreme environments raises fascinating questions about the adaptation of macromolecules and metabolic processes. Very low temperatures affect cells in many ways. Membranes lose their fluidity and are damaged by ice crystal formation. Chemical reactions and diffusion slow considerably. Proteins become too rigid to catalyze reactions and may undergo denaturation. At the opposite end of the temperature spectrum, heat denatures proteins and nucleic acids. Increased fluidity impairs metabolic processes in membranes. Some of the practical applications of the destructive effects of heat on microbes are sterilization by steam, pasteurization, and incineration of inoculating loops. Proteins in psychrophiles are, in general, rich in hydrophobic residues, display an increase in flexibility, and have a lower number of secondary stabilizing bonds when compared with homologous proteins from mesophiles. Antifreeze proteins and solutes that decrease the freezing temperature of the cytoplasm are common. The lipids in the membranes tend to be unsaturated to increase fluidity. Growth rates are much slower than those encountered at moderate temperatures. Under appropriate conditions, mesophiles and even thermophiles can survive freezing. Liquid cultures of bacteria are mixed with sterile glycerol solutions and frozen to −80 °C for long-term storage as stocks. Cultures can withstand freeze drying (lyophilization) and then be stored as powders in sealed ampules to be reconstituted with broth when needed.
Macromolecules in thermophiles and hyperthermophiles show some notable structural differences from what is observed in the mesophiles. The ratio of saturated to polyunsaturated lipids increases to limit the fluidity of the cell membranes. Their DNA sequences show a higher proportion of guanine–cytosine nitrogenous bases, which are held together by three hydrogen bonds in contrast to adenine and thymine, which are connected in the double helix by two hydrogen bonds. Additional secondary structures, ionic and covalent bonds, as well as the replacement of key amino acids to stabilize folding, contribute to the resistance of proteins to denaturation. The so-called thermoenzymes purified from thermophiles have important practical applications. For example, amplification of nucleic acids in the polymerase chain reaction (PCR) depends on the thermal stability of Taq polymerase , an enzyme isolated from T. aquaticus. Degradation enzymes from thermophiles are added as ingredients in hot-water detergents, increasing their effectiveness.
Check Your Understanding
- What temperature requirements do most bacterial human pathogens have?
- What DNA adaptation do thermophiles exhibit?
Eye on Ethics
Feeding the World…and the World’s Algae
Artificial fertilizers have become an important tool in food production around the world. They are responsible for many of the gains of the so-called green revolution of the 20th century, which has allowed the planet to feed many of its more than 7 billion people. Artificial fertilizers provide nitrogen and phosphorus, key limiting nutrients, to crop plants, removing the normal barriers that would otherwise limit the rate of growth. Thus, fertilized crops grow much faster, and farms that use fertilizer produce higher crop yields.
However, careless use and overuse of artificial fertilizers have been demonstrated to have significant negative impacts on aquatic ecosystems, both freshwater and marine. Fertilizers that are applied at inappropriate times or in too-large quantities allow nitrogen and phosphorus compounds to escape use by crop plants and enter drainage systems. Inappropriate use of fertilizers in residential settings can also contribute to nutrient loads, which find their way to lakes and coastal marine ecosystems. As water warms and nutrients are plentiful, microscopic algae bloom, often changing the color of the water because of the high cell density.
Most algal blooms are not directly harmful to humans or wildlife however, they can cause harm indirectly. As the algal population expands and then dies, it provides a large increase in organic matter to the bacteria that live in deep water. With this large supply of nutrients, the population of nonphotosynthetic microorganisms explodes, consuming available oxygen and creating “ dead zone s” where animal life has virtually disappeared.
Depletion of oxygen in the water is not the only damaging consequence of some algal blooms. The algae that produce red tides in the Gulf of Mexico, Karenia brevis , secrete potent toxins that can kill fish and other organisms and also accumulate in shellfish. Consumption of contaminated shellfish can cause severe neurological and gastrointestinal symptoms in humans. Shellfish beds must be regularly monitored for the presence of the toxins, and harvests are often shut down when it is present, incurring economic costs to the fishery. Cyanobacteria, which can form blooms in marine and freshwater ecosystems, produce toxins called microcystins , which can cause allergic reactions and liver damage when ingested in drinking water or during swimming. Recurring cyanobacterial algal blooms in Lake Erie (Figure 9.30) have forced municipalities to issue drinking water bans for days at a time because of unacceptable toxin levels.
This is just a small sampling of the negative consequences of algal blooms, red tides, and dead zones. Yet the benefits of crop fertilizer—the main cause of such blooms—are difficult to dispute. There is no easy solution to this dilemma, as a ban on fertilizers is not politically or economically feasible. In lieu of this, we must advocate for responsible use and regulation in agricultural and residential contexts, as well as the restoration of wetlands, which can absorb excess fertilizers before they reach lakes and oceans.