Bacterial Pathogenesis

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Sub-cause Age of staff Poor supervision 0 Lack of safety equipment Cause - Need for training/ refresher training Poor attitudes to safety Unsafe acts (lab coats, goggles, etc.) No approved laboratory methods or outdated methods in use Poor communication Change in method / management \ Accident Change in articles andsubstances handled InjUV Poor lighting Poor bench quality Poor storage of cultures No disinfection procedures b Figure 2.1. Multicausal analysis of a possible laboratory accident. Poor environmental conditions Unsafe conditions
References A review of a number of accidents occurring within microbiology lab- oratories (in the author's personal experience and observation) suggests that a number of similar underlying causes could be ascribed to the events. These may be presented as a multicausal analysis (Fig. 2.1), pro- viding a model of accident behavior. If staff wish to avoid a repetition of the mistakes of others, it is essential to address the underlying problems and not merely treat the 'symptoms of the disease' by reducing unsafe acts or conditions. This is part of risk management, described in more detail in Chapter 2.3. ACDP (1995). Categorisution of Biological Agents According to Hazard and Categories of Containment, 4th edn. Advisory Committee for Dangerous Pathogens, HSE Books, Sudbury, Suffolk. EC (1990). European Commission Directive 90/679/EEC establishing the protec- tion of workers from risks related to exposure to biological agents at work. Ofic. 1. Eur. Commun. L374,31.12.90. EC (1995). European Commission Directive 95/44/EC establishing the conditions under which certain harmful organisms, plants, plant products and other objects may be introduced into or moved within the community. Ofic. 1. Eur. Commun. LlW3.8.95. HSE (1978). Canvey: An lnvestigation of Potential Hazards from Operations in the Canvey lsZand/Thurrock Area. Health and Safety Executive Investigative Reports, HMSO, London. HSE (1992). The Tolerability of Risk from Nuclear Pmer Stations. Health and Safety Executive, Sheffield. WHO (1993). Laboratory Biosafety Manual, 2nd edn. World Health Organization, Geneva. 28
Page 2 and 3: Methods in Microbiology Volume 27
Page 4 and 5: Methods in M i cro bi ol ogy Volume
Page 6 and 7: Contents Contributors .............
Page 8 and 9: 6.10 Bacterial Exotoxins ..........
Page 10 and 11: Contributors Carlos F. Ambbile-Cuev
Page 12 and 13: Michele Farris Department of Bioche
Page 14 and 15: Mark Pallen Microbial Pathogenicity
Page 16 and 17: Preface 'I1 n'existe pas de science
Page 18 and 19: unknown (or unpredictable) function
Page 20 and 21: eflects a growing realization in re
Page 22 and 23: SECTION I Detection of Virulence Ge
Page 24 and 25: 1.1 Detection of Virulence Genes Ex
Page 26 and 27: permitting the simultaneous screeni
Page 28 and 29: tion have been used successfully to
Page 30 and 31: are impaired in their ability to su
Page 32 and 33: References most numerous facultativ
Page 34 and 35: SECTION 2 Laboratory Safety - Worki
Page 36 and 37: 2.1 Introduction Paul V. Tearle Nat
Page 38 and 39: Reference Thus, in both Chapters 2.
Page 40 and 41: 2.2 Safe Working Practices in the L
Page 42 and 43: it is the physical, noninfectious i
Page 44 and 45: difficulty of doing anything about
Page 46 and 47: elimination (1, the most effective)
Page 50 and 51: 2.3 Safe Working Practices in the L
Page 52 and 53: analysis and audit may also be used
Page 54 and 55: W w Second stage controls. . contro
Page 56 and 57: By-products Information References
Page 58 and 59: SECTION 3 Detection, Speciation and
Page 60 and 61: 3.1 Introduction Peter Vandamme Pos
Page 62 and 63: 3.2 Detection Margareta leven Assoc
Page 64 and 65: visualization of bacteria, but also
Page 66 and 67: days to weeks; many definitive test
Page 68 and 69: (NASBA) and the transcription media
Page 70 and 71: legionellae in respiratory specimen
Page 72 and 73: 3.3 Speciation Peter Vandamme Postd
Page 74 and 75: classdymg bacteria was analyzed. Am
Page 76 and 77: sequences indeed never show signifi
Page 78 and 79: 3.4 Identification Peter Vandamme P
Page 80 and 81: tional growth and metabolization of
Page 82 and 83: DNA sequence. Furthermore, 16s rRNA
Page 84 and 85: method suffice for numerous isolate
Page 86 and 87: List of Suppliers Polyphasic taxono
Page 88 and 89: SECTION 4 Host Interactions - Anima
Page 90 and 91: 4.1 Introduction T. J. Mitchell Div
Page 92 and 93: e examined histologically. Organ cu
Page 94 and 95: 4.2 Interaction of Bacteria and The
Page 96 and 97: The complexity of pathogenic mechan
Page 98 and 99:
++++++ COMPARISON OF DIFFERENT ORGA
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microscopy can assess cell damage (
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cient and deficient isogenic varian
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4.3 Transgenic and Knockout Animals
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(Hogan et al., 1994). The DNAmay be
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Knoc k-i n Animals In some experime
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etween two copies of the tamoxifen-
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g I Table 4.4. Examples of the use
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* Table 4.5. Examples of the use of
Page 116 and 117:
g m Table 4.5 continued IFN-y IFN-Y
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Numerous knockout mouse strains tha
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Gossen, M. and Bujard, H. (1992). T
Page 122 and 123:
Zinkernagel, R., Seimentz, M. and B
Page 124 and 125:
4.4 Interactions of Bacteria and Th
Page 126 and 127:
Route of Infection of cloned animal
Page 128 and 129:
Measure required length of intestin
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sampling, and be well tolerated by
Page 132 and 133:
Guerry, P., Pope, P. M., Burr, D. H
Page 134 and 135:
4.5 Interaction of Bacteria and The
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washed before resuspending in mediu
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the bacterial culture broth with th
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cells in suspension for several rea
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that bacterial cytotoxic factors or
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SECTION 5 Host I n te ract io ns -
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5.1 Introduction Michael J. Daniels
Page 148 and 149:
References bacteria often carry sin
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5.2 Testing Pathogenicity Michael J
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hosts than leaf spotting or wilt pa
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different plants can be infiltrated
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Table 5. I. The behavior of X. c pv
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Rudolph, K., Roy, M. A., Sasser, M.
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5.3 hup Genes and Their Function Al
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mental treatments. The timing and a
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essential positive regulators, or t
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of the Hrp system is presently obsc
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Dong, X., Mindrinos, M., Davis, K.
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5.4 Avirulence Genes Ulla Bonas lns
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of tissue due to disease developmen
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genes are tested for their plant in
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Bacteria expressing avirulence gene
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5.5 Extracellular Enzymes Their Rol
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- m Table 5.5. Extracellular enzyme
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+++a++ GENETIC DETERMINANTS OF EXTR
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Consequently, the precise link betw
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processing and so GspG, -H, -I, and
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++++++ ACKNOWLEDGEMENTS References
Page 190 and 191:
5.6 Bacterial Phytotoxins Carol L.
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ment. Since most bacterial phytotox
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ecently used in transgenic plants a
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Jones, J. D. G. and Gutterson, N. (
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5.7 Epiphytic Growth and Survival S
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++++++ EFFECTS OF SCALE ON SAMPLING
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References should be avoided so tha
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SECTION 6 Biochemical Approaches 6.
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6.1 Introduction: Fractionation of
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Gram-negative Bacteria Washed bacte
Page 210 and 211:
References trated lipoproteins, whi
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6.2 The Proteorne Approach C. David
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determining how its removal (e.g. b
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Spot Detection to one week before f
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MS acid residues in a polypeptide s
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An extension of the mass matching p
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approaches such as the construction
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infected with C. truchomutis, over
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6.3 Microscopic Approaches to the S
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The antigen of interest can be loca
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Figure 6.5. Whole-cell preparation
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Table 6.3. Pre-embedding immunolabe
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References Bacterial cell-associate
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6.4 Characterization of Bacterial S
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against a range of concentrations o
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single class of binding site on the
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Gel Filtration Whole cells are remo
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References This chapter has set out
Page 246 and 247:
List of Suppliers Wall, J., Ayoub,
Page 248 and 249:
6.5 Characterizing Flagella and Mot
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Phase-contrast microscopic observat
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of flagellar insertion as the ghost
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A last word of advice - do always c
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Tethered Cells Uniflagellate bacter
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Aizawa, $I., Dean, G. E., Jones, C.
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6.6 Fimbriae: Detection, Purificati
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Type 1 fimbriae, found on the major
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Detection of Fimbriae by Adhesive C
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~~~ Figure 6.16. Immunoelectron mic
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References The authors wish to than
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6.7 Isolation and Purification of C
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exopolysaccharides, EPSs) are relea
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cise chemistry of the LPS, and for
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Hitchcock-Brown whole-cell lysates
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Gotschlich, E. C., Fraser, 8. A., N
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6.8 Structural Analysis of Cell Sur
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26 I 3 Figure 6.18.1-D 'H NMR spect
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Composition Analysis Colorimetric a
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sugars with optically active alcoho
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Periodate oxidation Partial hydroly
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A 0 8H-lb.20 8 ti- IC. 40 Q ti- tc.
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the 6-deoxysugars, residues b and d
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References cal and NMR methods. The
Page 296 and 297:
List of Suppliers The following is
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6.9 Techniques for Analysis of Pept
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peptidoglycan isolation from these
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control and constant evaluation of
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Labeling Reaction and Sample Requir
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Billot-Klein, D., Gutmann, L., Sabl
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6.10 Bacterial Exotoxins Mahtab Moa
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ourselves to a summary of character
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(repeats in toxin) exotoxin family,
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Aeromonas hydrophila aerolysin, Ser
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1997). P. aeruginosa exotoxin A and
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Aktories, K. and Mohr, C. (1992). C
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Menestrina, G. F. (1991). Electroph
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6.11 A Systematic Approach to the S
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6 7 B 1 2 3 4 5 C 1 2 3 N-terminal
Page 326 and 327:
Analysis of Bacterial Cell Supernat
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tion as the components of the secre
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to attribute a role in secretion fo
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uninterpretable. A number of differ
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than one secretion machinery with v
Page 336 and 337:
References overexpression of a wild
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Merck, K. B., de Cock, H., Verheij,
Page 340 and 341:
6.12 Detection, Purification, and S
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an exogenous AHL (preferably OHHL)
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+++we SEPARATION, IDENTIFICATION AN
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lock the HPLC columns. Attention sh
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Figure 6.27. EI-MS fragmentation pa
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Synthesis of N-(3-oxoacyl)-~-homose
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6.13 Iron Starvation and Siderophor
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eceptor proteins form part of a hig
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phosphate-buffered systems can inte
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advantages both of speed and chemic
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produced by other bacterial and fun
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Ecker, D. J., Loomis, L. D., Cass,
Page 364 and 365:
SECTION 7 Molecular Genetic Approac
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7.1 Introduction Charles J. Dorman
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spectrum of difficulty lies Chlamyd
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7.2 Genetic Approaches to the Study
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transposon mutagenesis, which combi
Page 374 and 375:
Once a virulence gene has been iden
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References Alpuche-Aranda, C. M., S
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and virulence determinants in Vibri
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7.3 Signature Tagged MutagenesW * S
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sequence twice in a pool of 96 diff
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Table 7.4. Generation of the mutant
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Table 7.5. Colony blots 1. Grow the
Page 388 and 389:
we found that approximately 25% of
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References List of Suppliers primer
Page 392 and 393:
7.4 Physical Analysis of the Salmon
Page 394 and 395:
Table 7.8. Isolation of intact geno
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Cycle 2 Cycles 3 and 4 Frequently,
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0 2 4 6 8 10kb t 1 1 1 1 1 1 1 1 1
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1 2 3 4 5 I II Figure 7.3. (a) Auto
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List of Suppliers Liu, S.-L., Hesse
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7.5 Molecular Analvsis of Pseudomon
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of P. aeruginosa virulence genes ha
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Table 7.13. Murine’ models in use
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Table 7.14. Virulence of P. oerugin
Page 412 and 413:
Cash, H. A., Woods, D. E., McCullou
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Stieritz, D. D. and Holander, I. A.
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7.6 Molecular Genetics of Bordetell
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are under the control of a single g
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which may play a fundamental role i
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et al., 1988). On the contrary, the
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Figure 7.6. Schematic representatio
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Beier, D., Schwarz, B., Fuchs, T. M
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7.7 Genetic Manipulation of Enteric
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confers a catalase-positive phenoty
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esistance (Tc) (Sougakoff et al., 1
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Figure 7.7. (b) Schematic outline o
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Natural transformation strains test
Page 438 and 439:
References There has been rapid pro
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List of Suppliers of ferritin to ir
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7.8 Molecular Approaches for the St
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Table 7.22. Isolation of chromosoma
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Table 7.25. Protoplast transformati
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Colonies obtained after transformat
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References Table 7.29. RNA isolatio
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List of Suppliers Oxoid Monograph N
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7.9 Molecular Genetic Analvsis of S
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Table 7.30. Transposon mutagenesis
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(2) Linkage between the transposon
Page 460 and 461:
introduce point mutations generated
Page 462 and 463:
marker. The majority of bacteria th
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Resolution of Cointegrants by Trans
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performed using a fragment encompas
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transformation frequency is too low
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quency confines this procedure to c
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Bramley, A. J., Patel, A. H., O'Rei
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Novick, R. P. (1990). The staphyloc
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7.10 Molecular A P proaches to Stud
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DNA (Tan et al., 1995). Chlamydial
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was cloned from a pUC 8 expression
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factors have, so far, been unsucces
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tion of RNA polymerase core subunit
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SECTION 8 Gene txpression and Analy
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8.1 Gene Expression and Analysis Ia
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in multi-copy can titrate out possi
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permits direct targeting of putativ
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Northern blot analysis RT-PCR label
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accurately and in conjunction with
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DNA Footprinting This is a widely u
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(Frackman et al., 1990). It has bee
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Berg, C. M and Berg, D. E. (1996).
Page 504 and 505:
List of Suppliers Simpson, D. A. C.
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SECTION 9 Host Reactions - Animals
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9.1 Introduction Mark Roberts Depar
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in the case of septic shock. It is
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9.2 Strategies for the Studv of Int
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and saturable inhibition of bacteri
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adhesins. To facilitate the purific
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References We thank all the workers
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9.3 Cytoskeletal Rearrangements Ind
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(3) The interaction of intimin of e
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easy. For example, actin rearrangem
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Using mutated cell lines Inability
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Rosenshine, I., Donnenberg, M. S.,
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9.4 Activation and Inactivation of
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Actin Biochemistry guished. Intrace
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Figure 9.2. Schematic depiction of
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++++++ Rho-MODULATING BACTERIALTOXI
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Since the toxins are catalytically
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TcdB-I 0463 Tcd B-I 470 TcsL-82 48
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Effect on Bacterial Use of Actin Li
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small GTP-binding protein Rho regul
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tion of phorbol ester-stimulated ph
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9.5 Strategies for the Use of Gene
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+++a++ USE OF CYTOKINE AND CYTOKINE
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T Cell Proliferation Cytokine Studi
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Antibody Subclasses FACS Analysis S
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OCallaghan, D., Maskell, D., Liew,
Page 558 and 559:
SECTION 10 Host Reactions - Plants
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10.1 Host Reactions - Plants Charle
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Xanthomonas campestris pv. manihoti
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immunocytochemical studies within t
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Figure 10.2. Development of paramur
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etention and antibody recognition w
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Figure 10.4. Detection of phenolics
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Table 10.5. Histochemical detection
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Figure 10.6. Peroxidase activity in
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Figure 10.7. Localization of H,02 a
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A comparative summary of reactions
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Table 10.9. Preparation of gold pro
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symptom characteristic of infection
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Figure 10.10, Detection of flavanoi
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for conjugation via the hydroxy moi
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Figure 10.11 (A) Transmitted white
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References and protein/DNA complexe
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Buja, M., Eigenbrodt, M. L. and Eig
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Mansfield, J., Jenner, C., Hockenhu
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SECTION II Strategies and Problems
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11.1 Introduction Alejandro Craviot
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11.2 Strategies for Control of ComG
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endothelial cells to reach the subm
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The epidemiology of cholera is an i
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Table I I. I. Available animal mode
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acelular combinada con 10s toxoides
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11.3 Antibiotic Resistance and Pres
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(1) Enzymatic inactivation of the d
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ampicillin in these same strains in
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Single-event antibiotic usage may s
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SECTION I2 Internet Resources for R
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12.1 Internet Resources for Researc
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and DNA sequences (http://www.ncbi.
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++++++ SEQUENCE DATABASES The US Na
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searched as one of the options on t
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Web sites (e.g. httpJ/www.biochern.
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Index Accidents, 19,31 analysis, 27
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preliminary considerations, 232-233
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Exoproteins see Secreted proteins,
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Immunological detection of fimbriae
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tissue choice, 78 toxin measurement
Page 640 and 641:
identification, 20-21 pathogen cate
Page 642 and 643:
Transgenic animals, 83 in bacterial
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