Isolates ..................... 2, 6, 11–20, 23, 24, 26, 67, 70, 74,
85, 162, 279
Isolation ............................ 1, 4, 6, 8, 12, 18, 67, 71, 239,
247, 251, 267, 272, 278, 279, 283
K
Kidneys ...........................5, 42, 104, 105, 123, 156, 159,
167, 230, 249, 258, 263, 264, 268
L
Leptophages ..............................................................67–74
Leptospira .................. 1–9, 11–20, 23–28, 31–39, 41–50,
53–65, 67–74, 77–94, 97–106, 109, 110, 113,
115, 117, 123–127, 131–137, 139–148,
150–152, 161–164, 166, 169, 171, 188–190,
193–197, 200, 202, 204–206, 208, 209, 212,
213, 215–241, 243–245, 247–253, 257, 258,
260, 262, 263, 265–268, 271–274, 277, 279–281
Leptospira interrogans .....................vi, 16, 20, 23, 31, 32,
43, 53, 54, 60, 64, 77, 78, 97–106, 110, 115, 125,
150, 151, 159, 166, 173, 175, 188, 199, 200,
205, 208, 211–213, 225, 229–241, 245, 252,
258–260, 262, 264, 267, 272–274, 279
Leptospiral pathogenesis ..................................... 161, 172
Leptospires ..................... 1, 2, 4–6, 8, 25, 27, 31, 33–35,
37, 39, 57, 64, 67, 71, 99, 104, 117, 128,
140–142, 149–159, 166, 167, 169, 172, 188,
189, 192–196, 199, 200, 203, 221, 229–231,
234, 235, 239, 243–245, 247, 248, 250, 253,
257, 258, 267, 271, 273, 277–279
Lipopolysaccharide (LPS) ...................... 53–64, 147, 148,
195–197, 265, 271, 273
Luciferases ............................................................ 150, 159
M
Macrophages ......................................... 41, 187, 199–206
MALDI-ToF MS.......................................................23, 24
Microscopic agglutination test (MAT) .......................280,
281, 283
Motilities........................... 31, 78, 82, 91, 121, 123–128,
131, 140–143, 145–147, 231
Mutagenesis.........................................78, 80, 87, 97, 109
N
Nested PCR................................ 80, 86–88, 93, 102, 279
96-well plate ...............................213, 222, 226, 227, 263
Nucleic acid amplification test (NAAT).............. 279–281
P
Pathogenesis ........................................................... 78, 162
Phagosome maturation........................................ 199, 206
Phase contrast images ................................. 208, 209, 211
Phenol-chloroform extraction........................................ 42
Phenol extraction ..................................54, 57, 60, 62, 63
Plating.................................. 67, 71, 80, 83, 90, 225, 226
Polarized-monolayers ..................................162, 167–169
Polyethylene glycol (PEG) precipitation .......... 67, 69, 70
Propidium iodide .......................................................... 217
Proteases .............................................. 188–190, 194–196
Proteinase k. ........................................................... 55, 201
Q
Quantification ............................... 31, 41, 42, 58, 63, 64,
129, 156, 157, 207–214, 222, 244, 247–251, 263
R
Recombinant proteins....................... 172, 177, 178, 180,
258, 265, 281
Resazurin ........................... 216, 217, 220, 222, 226, 227
Resorufin .............................................216, 222, 226, 227
RIN ..................................................................... 43, 47, 50
RNA ..............................41–50, 110, 113, 239, 244, 246,
247, 249, 251, 280
RNA-Seq ...............................................41, 43, 46, 47, 49
RT-PCR ............................................................43, 46, 280
S
Selective agents.................................................................. 4
Serology ..........43, 84, 91, 219, 220, 233–236, 279–283
Serum susceptibility ...................................................... 188
Silver staining ........................................ 56, 57, 60–62, 64
Single-guide RNA (sgRNA).....110, 111, 113, 115, 116,
119, 120
Soils ................ 6, 8, 32, 42, 74, 123, 229, 243, 271–273
Spirochetes .......................32, 53, 78, 97, 109, 123, 131,
139, 146, 161, 193, 243, 244, 264, 272, 273
Survival ...............41, 193, 200, 215, 221–226, 263, 271
SYTO
® .....................32–34, 38, 217, 220, 221, 223–225
T
Taxonomy........................................................................ 23
TEM.................................................................... 67, 71–74
Thiocyanate ........................................................ 42, 44, 48
Translocation ........................................................ 161–170
Transposon mutant library .......................................77–94
Transposons................. 42, 77–94, 97–99, 104, 150, 231
Transwell
® ................................................... 162, 166, 168
U
Urine....................5, 8, 42, 90, 123, 154, 156, 157, 159,
229, 230, 243, 248, 257, 273, 278–280, 283
V
Vaccines................................................243, 244, 257–268
Videomicroscopy..........................................123–125, 127
Virulence factors ................... 77, 97, 140, 244, 273, 274
290
LEPTOSPIRA SPP.: METHODS AND PROTOCOLS
Index
85, 162, 279
Isolation ............................ 1, 4, 6, 8, 12, 18, 67, 71, 239,
247, 251, 267, 272, 278, 279, 283
K
Kidneys ...........................5, 42, 104, 105, 123, 156, 159,
167, 230, 249, 258, 263, 264, 268
L
Leptophages ..............................................................67–74
Leptospira .................. 1–9, 11–20, 23–28, 31–39, 41–50,
53–65, 67–74, 77–94, 97–106, 109, 110, 113,
115, 117, 123–127, 131–137, 139–148,
150–152, 161–164, 166, 169, 171, 188–190,
193–197, 200, 202, 204–206, 208, 209, 212,
213, 215–241, 243–245, 247–253, 257, 258,
260, 262, 263, 265–268, 271–274, 277, 279–281
Leptospira interrogans .....................vi, 16, 20, 23, 31, 32,
43, 53, 54, 60, 64, 77, 78, 97–106, 110, 115, 125,
150, 151, 159, 166, 173, 175, 188, 199, 200,
205, 208, 211–213, 225, 229–241, 245, 252,
258–260, 262, 264, 267, 272–274, 279
Leptospiral pathogenesis ..................................... 161, 172
Leptospires ..................... 1, 2, 4–6, 8, 25, 27, 31, 33–35,
37, 39, 57, 64, 67, 71, 99, 104, 117, 128,
140–142, 149–159, 166, 167, 169, 172, 188,
189, 192–196, 199, 200, 203, 221, 229–231,
234, 235, 239, 243–245, 247, 248, 250, 253,
257, 258, 267, 271, 273, 277–279
Lipopolysaccharide (LPS) ...................... 53–64, 147, 148,
195–197, 265, 271, 273
Luciferases ............................................................ 150, 159
M
Macrophages ......................................... 41, 187, 199–206
MALDI-ToF MS.......................................................23, 24
Microscopic agglutination test (MAT) .......................280,
281, 283
Motilities........................... 31, 78, 82, 91, 121, 123–128,
131, 140–143, 145–147, 231
Mutagenesis.........................................78, 80, 87, 97, 109
N
Nested PCR................................ 80, 86–88, 93, 102, 279
96-well plate ...............................213, 222, 226, 227, 263
Nucleic acid amplification test (NAAT).............. 279–281
P
Pathogenesis ........................................................... 78, 162
Phagosome maturation........................................ 199, 206
Phase contrast images ................................. 208, 209, 211
Phenol-chloroform extraction........................................ 42
Phenol extraction ..................................54, 57, 60, 62, 63
Plating.................................. 67, 71, 80, 83, 90, 225, 226
Polarized-monolayers ..................................162, 167–169
Polyethylene glycol (PEG) precipitation .......... 67, 69, 70
Propidium iodide .......................................................... 217
Proteases .............................................. 188–190, 194–196
Proteinase k. ........................................................... 55, 201
Q
Quantification ............................... 31, 41, 42, 58, 63, 64,
129, 156, 157, 207–214, 222, 244, 247–251, 263
R
Recombinant proteins....................... 172, 177, 178, 180,
258, 265, 281
Resazurin ........................... 216, 217, 220, 222, 226, 227
Resorufin .............................................216, 222, 226, 227
RIN ..................................................................... 43, 47, 50
RNA ..............................41–50, 110, 113, 239, 244, 246,
247, 249, 251, 280
RNA-Seq ...............................................41, 43, 46, 47, 49
RT-PCR ............................................................43, 46, 280
S
Selective agents.................................................................. 4
Serology ..........43, 84, 91, 219, 220, 233–236, 279–283
Serum susceptibility ...................................................... 188
Silver staining ........................................ 56, 57, 60–62, 64
Single-guide RNA (sgRNA).....110, 111, 113, 115, 116,
119, 120
Soils ................ 6, 8, 32, 42, 74, 123, 229, 243, 271–273
Spirochetes .......................32, 53, 78, 97, 109, 123, 131,
139, 146, 161, 193, 243, 244, 264, 272, 273
Survival ...............41, 193, 200, 215, 221–226, 263, 271
SYTO
® .....................32–34, 38, 217, 220, 221, 223–225
T
Taxonomy........................................................................ 23
TEM.................................................................... 67, 71–74
Thiocyanate ........................................................ 42, 44, 48
Translocation ........................................................ 161–170
Transposon mutant library .......................................77–94
Transposons................. 42, 77–94, 97–99, 104, 150, 231
Transwell
® ................................................... 162, 166, 168
U
Urine....................5, 8, 42, 90, 123, 154, 156, 157, 159,
229, 230, 243, 248, 257, 273, 278–280, 283
V
Vaccines................................................243, 244, 257–268
Videomicroscopy..........................................123–125, 127
Virulence factors ................... 77, 97, 140, 244, 273, 274
290
LEPTOSPIRA SPP.: METHODS AND PROTOCOLS
Index