Growth Measurement

Arthur L. Koch

doi:10.1128/9781555817497.ch9

Chapter 9 : Growth Measurement

Author: Arthur L. Koch

Content Type: Reference

Publication Year: 2007

Category: Microbial Genetics and Molecular Biology

Book DOI: 10.1128/9781555817497

Chapter DOI: 10.1128/9781555817497.ch9

MyBook is a cheap paperback edition of the original book and will be sold at uniform, low price.

Preview this chapter:

Growth Measurement, Page 1 of 2

< Previous page | Next page > /docserver/preview/fulltext/10.1128/9781555817497/9781555812232_Chap09-1.gif /docserver/preview/fulltext/10.1128/9781555817497/9781555812232_Chap09-2.gif

Abstract:

This chapter talks about four classes of pitfalls in bacterial growth measurement. The change of enteric bacteria from large RNA-rich forms in the exponential phase to small RNA-poor forms in the stationary phase has many of the aspects of differentiation. Evidence of the difficulties involved is the fact that many of the people who helped develop the technique for measuring distribution of cell volumes no longer use it. This article, therefore, only presents the principles, mentions the difficulties and the attempted solutions to these difficulties, and directs the reader to published literature. Three colony count methods are discussed. The first is the spread plate, in which all colonies are surface colonies. The second method is the layered plate. The third method is the pour plate. Flow cytometry has become an extremely powerful method for the studies of many aspects of the biology of eucaryotes, but the methods are only now coming into their own in the study of the biology of procaryotes. The concentration of viable cells can be roughly estimated by the most-probable-number (MPN) method. For growth measurements, the time can be precisely defined, and the major error is in the measurement of the number of cells or other indices of biomass.

Citation: Koch A. 2007. Growth Measurement, p 172-199. In Reddy C, Beveridge T, Breznak J, Marzluf G, Schmidt T, Snyder L (ed), Methods for General and Molecular Microbiology, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817497.ch9

Highlighted Text: Show | Hide

Full text loading...

Figures

Growth Measurement

[com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/author/10.1128/9781555817497.chap9-1,webId=/content/author/10.1128/9781555817497.chap9-1,properties={foaf_givenname=Arthur L., foaf_name=Arthur L. Koch, foaf_surname=Koch}]]

/content/10.1128/9781555817497.chap9.fig9-1

FIGURE 1

Spreaders. The one at the top is bent from a paper clip and fitted into a length of Teflon tubing. The one at the bottom is made by fusing the end of a Pasteur pipette and then bending it sharply in two places by using a Bunsen burner with a wing tip to define the spreading region. The handles of both are bent to the convenience of the user. Both spreaders have low heat capacity. The spreader at the top absorbs fewer bacteria because of its Teflon construction.

10.1128/9781555817497/fig9-1_thmb.gif

10.1128/9781555817497/fig9-1.gif

FIGURE 1

/content/10.1128/9781555817497.chap9.fig9-2

FIGURE 2

Errors that arise in using the MPN method. The CV for a single level of dilution is shown as a function of the percentage of sterile tubes (bottom abscissa) and the mean number of cells per tube (top abscissa). The respective optima occur at 20.8% and 1.59 mean cells per tube for a varying number of tubes (n) prepared at a single dilution level.

10.1128/9781555817497/fig9-2_thmb.gif

10.1128/9781555817497/fig9-2.gif

FIGURE 2

/content/10.1128/9781555817497.chap9.fig9-3

FIGURE 3

Typical water sorption isotherm curve for bacterial cells.

10.1128/9781555817497/fig9-3_thmb.gif

10.1128/9781555817497/fig9-3.gif

FIGURE 3

Typical water sorption isotherm curve for bacterial cells.

/content/10.1128/9781555817497.chap9.fig9-4

FIGURE 4

Light-scattering patterns from randomly oriented bacteria showing the angular distribution of light scattered by a beam traversing from left to right through a suspension of cells with the size and physical properties of Escherichia coli grown in minimal medium. The upper pattern shows the distribution if the cells are ellipsoidal with an axial ratio of 4:1. The lower pattern shows the distribution for spherical cells.

10.1128/9781555817497/fig9-4_thmb.gif

10.1128/9781555817497/fig9-4.gif

FIGURE 4

/content/10.1128/9781555817497.chap9.fig9-5

FIGURE 5

Schematic designs for filter-type “colorimeter” instruments. (A) Narrow-beam instrument. Almost all the scattered light deviates enough to escape falling on the photodetector. (B) Wide-beam instrument. A variable fraction of the scattered light falls on the detector, and thus the sensitivity is lowered.

10.1128/9781555817497/fig9-5_thmb.gif

10.1128/9781555817497/fig9-5.gif

FIGURE 5

/content/10.1128/9781555817497.chap9.fig9-6

FIGURE 6

Absorbance (at 420 nm) as a function of bacterial cell concentration. The dashed line shows the theoretical relationship. The solid line shows an actual result obtained with 15 dilutions of a bacterial culture, whose concentrations were measured by dry-weight determinations and whose plot was obtained by the best-fitting quadratic curve that was forced to pass through the origin.

10.1128/9781555817497/fig9-6_thmb.gif

10.1128/9781555817497/fig9-6.gif

FIGURE 6

/content/10.1128/9781555817497.chap9.fig9-7

FIGURE 7

Growth curve plotted on semilogarithmic paper. See the text for conditions. The horizontal arrows are drawn to the curve at values of bacterial concentrations that are convenient powers of 2 apart, which facilitates estimations of the doubling time.

10.1128/9781555817497/fig9-7_thmb.gif

10.1128/9781555817497/fig9-7.gif

FIGURE 7

/content/10.1128/9781555817497.chap9.fig-pg195-1a

Untitled

10.1128/9781555817497/fig-pg195-1a_thmb.gif

10.1128/9781555817497/fig-pg195-1a.gif

Untitled

/content/10.1128/9781555817497.chap9.fig-pg195-1b

Untitled

10.1128/9781555817497/fig-pg195-1b_thmb.gif

10.1128/9781555817497/fig-pg195-1b.gif

Untitled

References

/content/book/10.1128/9781555817497.chap9

1. Dawson, P. S. S. (ed.). 1974. Microbial Growth. Halsted Press, New York, NY. A collection of papers that earlier dominated the study of physiology of bacterial growth..

2. Gall, L. S.,, and W. A. Curby. 1979. Instrumental Systems for Microbiological Analysis of Body Fluids. CRC Press, Inc., Boca Raton, FL.

3. Kavenagh, F. (ed.). 1963. Analytical Microbiology, vol. 1. Academic Press, Inc., New York, NY.

4. Kavenagh, F. (ed.). 1972. Analytical Microbiology, vol. 2. Academic Press, Inc., New York, NY.

5. Meadows, P.,, and S. J. Pirt (ed.). 1969. Microbial growth. Symp. Soc. Gen. Microbiol. 19: 1– 450.

6. Meynell, G. G.,, and E. Meynell. 1970. Theory and Practice in Experimental Bacteriology, 2nd ed. Cambridge Press, Cambridge, United Kingdom.

7. Miller, J. H. 1972. Experiments in Molecular Genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.

8. Pirt, S. J. 1975. Principles of Microbe and Cell Cultivation. Blackwell Scientific Publications, Oxford, United Kingdom.

9. Amann, R. I.,, B. J. Binder,, R. J. Olson,, S. W. Chisholm,, R. Devereux,, and D. A. Stahl. 1990. Combination of 16S rRNA targeted oligonucleotide probes with flow cytometry for analyzing mixed microbial populations. Appl. Environ. Microbiol. 56: 1919– 1925.

10. Black, S. H.,, and P. Gerhardt. 1962. Permeability of bacterial spores. IV. Water content, uptake, and distribution. J. Bacteriol. 83: 960– 987.

11. Bridges, B. A. 1976. Survival of bacteria following exposure to ultraviolet and ionizing radiation. Symp. Soc. Gen. Microbiol. 26: 183– 208.

12. Brock, T. D. 1967. Bacterial growth in the sea; direct analysis by thymidine autoradiography. Science 155: 81– 83.

13. Brock, T. D. 1971. Microbial growth rates in nature. Bacteriol. Rev. 35: 39– 58.

14. Buchanan, R. E. 1918. Life phases in a bacterial culture. J. Infect. Dis. 23: 109– 125. This classic has been reprinted in reference 1..

15. Button, D. K.,, and B. R. Robertson. 1989. Kinetics of bacterial processes in natural aquatic systems based on biomass determined by high-resolution flow cytometry. Cytometry 10: 558– 563.

16. Calam, C. T., 1969. The evaluation of mycelial growth, p. 567– 591. In J. R. Norris, and D. W. Ribbons (ed.), Methods in Microbiology, vol. 1. Academic Press, Inc., New York, NY.

17. Campbell, A. 1957. Synchronization of cell division. Bacteriol. Rev. 21: 263– 272.

18. Capaldo, F. N.,, and S. D. Barbour,. 1975. The role of the rec genes in the viability of Escherichia coli KI2, p. 405– 418. In P. C. Hanawalt, and R. B. Setlow (ed.), Molecular Mechanisms for Repair of DNA, part A. Plenum Publishing Corp., New York, NY.

19. Cato, E. P.,, C. S. Cufflins,, L. V. Holdeman,, J. L. Johnson,, W. E. C. Moore,, R. M. Smibert,, and K. D. S. Smith. 1970. Outline of Culture Methods in Anaerobic Bacteriology. Virginia Polytechnic Institute and State University, Blacksburg.

20. Clifton, C. E. 1966. Aging of Escherichia coli. J. Bacteriol. 92: 905– 912.

21. Dicker, D. T.,, and M. L. Higgins. 1987. Cell cycle changes in the buoyant density of exponential-phase cells of Streptococcus faecium. J. Bacteriol. 169: 1200– 1204.

22. Drake, J. F.,, and H. M. Tsuchlya. 1973. Differential counting in mixed cultures with Coulter counters. Appl. Microbiol. 26: 9– 13.

23. Edwards, C.,, J. Porter,, J. R. Saunders,, J. Diaper,, J. A. W. Morgan,, and R. W. Pickup. 1992. Flow cytometry and microbiology. SGM Q. 19: 105– 108.

24. Finney, D. J. 1964. Statistical Method in Biological Assay, 2nd ed., p. 570– 586. Hafner Publishing Co., New York, NY.

25. Gray, T. R. G. 1976. Survival of vegetative microbes in soil. Symp. Soc. Gen. Microbiol. 26: 327– 364.

26. Guerrero, R.,, C. Pedros-Alió,, T. M. Schmidt,, and J. Mas. 1985. A survey of buoyant density of microorganisms in pure cultures and natural samples. Microbiologia 1: 53– 65.

27. Gunther, H. H.,, and F. Berhgter. 1971. Bestimmung der Trockenmasse von Zellsuspensionen durch Extinktions-messungen. Z. Allg. Mikrobiol. 11: 191– 197.

28. Halvorson, H. O.,, and N. R. Ziegler. 1933. Application of statistics to problems in bacteriology. 1. A means of determining bacterial population by the dilution method. J. Bacteriol. 25: 101– 121.

29. Heden, C.-G.,, and T. Illeni. 1975. Automation in Microbiology and Immunology. John Wiley & Sons, Inc., New York, NY.

30. Holdernan, L. V.,, E. P. Cato,, and W. E. C. Moore (ed.). 1977. Anaerobe Laboratory Manual, 4th ed. Virginia Polytechnic Institute and State University, Blacksburg.

31. Hungate, R. E., 1971. A roll tube method for cultivation of strict anaerobes, p. 117– 132. In J. R. Norris, and D. W. Ribbons (ed.), Methods in Microbiology, vol. 3a. Academic Press, Inc., New York, NY.

32. Kavenagh, F., 1972. Photometric assaying, p. 43– 121. In F. Kavenagh (ed.), Analytical Microbiology. Academic Press, Inc., New York, NY.

33. Kerker, M.,, D. D. Coke,, H. Chew,, and P. J. McNulty. 1978. Light scattering by structural spheres. J. Opt. Soc. Am. 68: 592– 601.

34. Koch, A. L. 1961. Some calculations on the turbidity of mitochondria and bacteria. Biochim. Biophys. Acta 51: 429– 441.

35. Koch, A. L. 1968. Theory of angular dependence of light scattered by bacteria and similar-sized biological objects. J. Theor. Biol. 18: 133– 156.

36. Koch, A. L. 1970. Turbidity measurements of bacterial cultures in some available commercial instruments. Anal. Biochem. 38: 252– 259.

37. Koch, A. L. 1971. The adaptive responses of Escherichia coli to a feast and famine existence. Adv. Microb. Physiol. 6: 147– 217.

38. Koch, A. L. 1975. Lag in adaptation to lactose as a probe to the timing of permease incorporation into the cell membrane. J. Bacteriol. 124: 435– 444.

39. Koch, A. L. 1975. The kinetics of mycelial growth. J. Gen. Microbiol. 89: 209– 216.

40. Koch, A. L. 1976. How bacteria face depression, recession, and derepression. Perspect. Biol. Med. 20: 44– 63.

41. Koch, A. L.,, and E. Ehrenfeld. 1968. The size and shape of bacteria by light scattering measurements. Biochim. Biophys. Acta 165: 262– 273.

42. Koch, A. L.,, and G. H. Gross. 1979. Growth conditions and rifampin susceptibility. Antimicrob. Agents Chemother. 15: 220– 228.

43. Kubitschek, H. E. 1964. Apertures for Coulter counters. Rev. Sci. Instrum. 35: 1598– 1599.

44. Kubitschek, H. E., 1969. Counting and sizing microorganisms with the Coulter counter, p. 593– 610. In J. R. Norris, and D. W. Ribbons (ed.), Methods in Microbiology, vol. 1. Academic Press, Inc., New York, NY.

45. Kubitschek, H. E. 1987. Buoyant density variation during the cell cycle in microorganisms. Crit. Rev. Microbiol. 14: 73– 97.

46. Maaløe, O.,, and N. O. Kjeldgaard. 1966. Control of Macromolecular Synthesis. W. A. Benjamin, Inc., New York, NY.

47. Mallette, M. F., 1969. Evaluation of growth by physical and chemical means, p. 521– 566. In J. R. Norris, and D. W. Ribbons (ed.), Methods in Microbiology, vol. 1. Academic Press, Inc., New York, NY.

48. Marquis, R. E.,, and P. Gerhardt. 1959. Polymerized organic salts of sulfonic acid used as dispersing agents in microbiology. Appl. Microbiol. 7: 105– 108.

49. Marshall, R. T., 1993. Media, p. 89– 101. In R. T. Marshall (ed.), Standard Methods for the Examination of Dairy Products, 16th ed. American Public Health Association, Washington, DC.

50. Melamed, M. R.,, T. Lindmo,, and M. L. Mendelsohn (ed.). 1990. Flow Cytometry and Sorting, 2nd ed. Wiley- Liss, New York, NY.

51. Mitruka, B. M. 1976. Methods of Detection and Identification of Bacteria. CRC Press, Inc., Cleveland, OH.

52. Nir, R.,, Y. Yisraeli,, R. Lamed,, and E. Sajar. 1990. Flow cytometry sorting of viable bacteria and yeasts according to O-galactosidase activity. Appl. Environ. Microbiol. 56: 3861– 3866.

53. Norris, K. P.,, and E. O. Powell. 1961. Improvements in determining total counts of bacteria. J. R. Microsc. Soc. 80: 107– 119.

54. Novick, A.,, and L. Szilard. 1951. Experiments with the chemostat on the spontaneous mutations of bacteria. Proc. Natl. Acad. Sci. USA 36: 708– 719.

55. Ormerod, M. G. (ed.). 2000. Flow Cytometry: a Practical Approach, 3rd ed. Oxford University Press, New York, NY.

56. Perfiliv, B. V.,, and D. R. Gabe. 1969. Capillary Methods of Investigation of Microorganisms. University of Toronto Press, Toronto, Canada.

57. Postgate, J. R. 1967. Viability measurements and the survival of microbes under minimal stress. Adv. Microb. Physiol. 1: 2– 23.

58. Postgate, J. R., 1969. Viable counts and viability, p. 611– 628. In J. R. Norris, and D. W. Ribbons (ed.), Methods in Microbiology, vol. 1. Academic Press, Inc., New York, NY.

59. Ryan, F. J.,, G. W. Beadle,, and E. L. Tatum. 1943. The tube method of measuring the growth rate of neurospora. Am. J. Bot. 30: 784– 799.

60. Schmidt, E. L.,, R. O. Bantkole,, and B. B. Bohlool. 1968. Fluorescent-antibody approach to study of rhizobia in soil. J. Bacteriol. 95: 1987– 1992.

61. Shapiro, H. M. 2003. Practical Flow Cytometry, 4th ed. Wiley-Liss, New York, NY.

62. Shehata, T. E.,, and A. G. Marr. 1971. Effect of nutrient concentration on the growth of Escherichia coli. J. Bacteriol. 107: 210– 216.

63. Shuler, M. L.,, R. Arls,, and H. M. Tsuchlya. 1972. Hydrodynamic focusing and electronic cell-sizing techniques. Appl. Microbiol. 24: 384– 388.

64. Sokal, R. R.,, and F. J. Rohlf. 1969. Biometry. W. H. Freeman & Co., San Francisco, CA.

65. Spielman, L.,, and S. L. Goren. 1968. Improving resolution in Coulter counting by hydrodynamic focusing. J. Colloid Interface Sci. 26: 175– 182.

66. Steen, H. B., 1990. Flow cytometric studies of microorganisms, p. 605– 622. In M. R. Melamed,, T. Linmo,, and M. L. Mendelsohn (ed.), Flow Cytometry and Sorting, 2nd ed. Wiley-Liss, New York, NY.

67. Stock, J. B.,, B. Rauch,, and S. Roseman. 1977. Periplasmic spaces in Salmonella typhimurium and Escherichia coli. J. Biol. Chem. 252: 7850– 7861.

68. Taney, T. A.,, J. R. Gerke,, and J. F. Pagano. 1963. Automation of microbiological assays, p. 219– 247. In F. Kavenagh (ed.), Analytical Microbiology. Academic Press, Inc., New York, NY.

69. Tisa, L. S.,, T. Koshikawa,, and P. Gerhardt. 1982. Wet and dry bacterial spore densities determined by buoyant sedimentation. Appl. Environ. Microbiol. 43: 1307– 1310.

70. Troller, J. A.,, and J. H. B. Christian. 1978. Water Activity and Food. Academic Press, Inc., New York, NY.

71. VanDilla, M. A.,, P. N. Dean,, O. D. Laerum,, and M. R. Melamed. 1985. Flow Cytometry: Instrumentation and Data Analysis. Academic Press, Orlando, FL.

72. Vollenwelder, R. A. 1974. A Manual on Methods for Measuring Primary Production in Aquatic Environments. IBP handbook no. 12. Blackwell Scientific Publications, Oxford, United Kingdom.

73. Wyatt, P. J., 1973. Differential light scattering techniques for microbiology, p. 183– 263. In J. R. Norris, and D. W. Ribbons (ed.), Methods in Microbiology, vol. 8. Academic Press, Inc., New York, NY.

74.Wyatt Technology. 1988. Literature on Model F and Model B Dawn Instruments. Wyatt Technology, Santa Barbara, CA.

75. Zimmerman, U.,, J. Schultz,, and G. Pilwat. 1973. Transcellular ion flow in Escherichia coli B and electrical sizing of bacterias. Biophys. J. 13: 1005– 1013.

Tables

Growth Measurement

/content/10.1128/9781555817497.chap9.tab9-1

TABLE 1

Sample data for exponential growth by graphical estimation or pocket calculator computation

10.1128/9781555817497/tab9-1_thmb.gif

10.1128/9781555817497/tab9-1.gif

TABLE 1

Sample data for exponential growth by graphical estimation or pocket calculator computation

/content/10.1128/9781555817497.chap9.ut09-1

Untitled

SAMPLE DATA FOR MPN COMPUTATION

10.1128/9781555817497/tab-pg195-1_thmb.gif

10.1128/9781555817497/tab-pg195-1.gif

Untitled

SAMPLE DATA FOR MPN COMPUTATION

/content/10.1128/9781555817497.chap9.tab-pg197-1

Untitled

10.1128/9781555817497/tab-pg197-1_thmb.gif

10.1128/9781555817497/tab-pg197-1.gif

Untitled

/content/10.1128/9781555817497.chap9.tab-pg197-2

Untitled

10.1128/9781555817497/tab-pg197-2_thmb.gif

10.1128/9781555817497/tab-pg197-2.gif

Untitled

/content/10.1128/9781555817497.chap9.tab9-A1

TABLE A1

Sample data and output, E. coli ML308, 28°C, M9 Med, 10/25/2003, Run B

10.1128/9781555817497/tab9-A1_thmb.gif

10.1128/9781555817497/tab9-A1.gif

TABLE A1

Sample data and output, E. coli ML308, 28°C, M9 Med, 10/25/2003, Run B