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Manual of Methods of Pure Culture Study

ISBN : 9789385961069

Author(s) : Abhay B. Solunke, Venkat S. Hamde, Prashant S. Wakte, Rajendra S. Awasthi

Year : 2015

Edition : 1

Cover Type : Hard Cover

Size : 9.5 x 6.5

Weight : 610 gr./327P

Publisher : NPI

Subject : Microbiology

Language : English

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Dr. Abhay B. Solunke

Head Department of Microbiology,

Shri Govindrao Munghate Arts& Science College,

Kurkheda.441  209

Dist. Gadchiroli (Maharashtra)

(Dr. Abhay B. Solunke did his M.Sc. from Marathwada University Aurangabad in 1993. He is recognised University PG teachers in Microbiology and Biotechnology of Swami Ramanand Teerth Marathwada University, Nanded and has taught devotedly at undergraduate and postgraduate level. He has several research papers and projects to his credit and also authored books in his field of study. He has introduced M.Sc. Industrial Microbiology as innovative subject in UGC X plan programme. His Ph.D. work is associated with Mushrooms and the bioconversion of agro-wastes. He is also Ph.D. Guide in Microbiology and Biotechnology of Gondwana University, Gadchiroli. He is also Chairman of Board of studies in Microbiology and Biotechnology and Member of Academic Council Gondwana University, Gadchiroli. He is currently serving as Head Department of Microbiology and IQAC coordinator at Shri Govindrao Munghate Arts & Science College, Kurkheda. Dist. Gadchiroli. )

 

Dr. Venkat S. Hamde

Department of Microbiology,

Yogeshwari Mahavidyalaya,Ambajogai

Dist. Beed(Maharashtra)

 

Dr. Prashant S. Wakte

Head Department of Microbiology,

DSM College, Parbhani. 431 401(MS)

 

Dr. Rajedra  S. Awasthi

Head Department of Microbiology,

Shivaji College, Renapur. 413 527(Maharashtrs)

This manual is intended for use in those types of microbiological work which involve the study of microbial cultures or of viruses, either for identification or for learning the properties of the  organisms investigated.

This Manual is intended for use in that type of microbiological work known as "pure culture study of bacteria".

This book takes the place of the loose-leaf publication issued during the period of 1923-2013 under the name of ''Manual of Methods For Pure Culture Study." The present manual covers a wider scope.

The standardization of methods tends to hinder the development of new techniques, while the chief function of this manual is to stimulate its development.

The methods in this manual, therefore, are merely claimed to be those regarded as satisfactory.

A thorough course of laboratory instruction in Microbiology is absolutely essential to the proper education of the student of the present day. The practical knowledge thus acquired in the methods of handling bacteria, in the precautions necessary to the prevention of personal infection, and in the methods for the recognition and for the destruction of disease-producing organisms is fundamental and invaluable. Such information is directly useful

as a means of diagnosis; it is necessary to the successful performance of antiseptic operations and is indispensable to the proper execution and understanding of the common hygienic measures for the prevention of communicable diseases.

It is therefore evident that the course in Microbiology should not be inferior, either in length or in the character of the instruction, to any other laboratory course offered in the Microbiology curriculum. The student should be taught to work, not merely with a few harmless bacteria, but especially with all of the common pathogenic organisms. The exclusion of the latter organisms from a laboratory course on the plea of danger is an admission of weakness in instruction or in supervision.

 


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Sr. No.

Title

Page No.

1

Media and Cultural Techniques

 

1.1

Introduction to Culture Media, Stains and Solutions.

 

1.2

Preparation of Culture Media

 

1.3

Discovery of Agar

 

1.4

MacConkey agar

 

1.5

Cotton Plug

 

1.6

Petri Plates

 

1.7

Reinforced Clostridial Medium

 

1.8

Agarose

 

1.9

Beef Extract

 

1.10

Malt Extract

 

1.11

Peptone

 

1.12

Meat Peptones

 

1.13

Casein Peptones

 

1.14

Soy Peptones

 

1.15

Casein Hydrolysate

 

1.16

Lactoalbumin Hydrolysate

 

1.17

Malt Extract

 

1.18

Mycological Peptone

 

1.19

Peptonized Milk

 

1.20

Proteose Peptone

 

1.21

Sheep Blood, Defibrinated

 

1.22

Tryptone

 

1.23

Tryptose

 

1.24

Yeast Extract

 

1.25

Biphasic media

 

1.26

Streak Plate Method

 

1.27

Autoclaving

 

1.28

pH Indicators

 

1.29

Water Activity

 

1.30

Dyes for Culture Media

 

1.31

Different Categories of Culture Media

 

1.32

Media and Purpose

 

1.33

Storage of culture media

 

1.34

Quantal microbiology

 

1.35

Bacterial Colony Characteristics

 

1.36

Meaning of pure culture study

 

1.37

History of Methods for Pure Culture Study

 

1.38

Determining the characteristics of a culture

 

1.39

Relation to Taxonomy

 

1.40

Media used in pure culture study

 

1.41

Natural storage media

 

1.42

Indicator media

 

1.43

Use of Methylene Blue as Indicator of Anaerobiosis

 

1.44

Use of aerobe to absorb oxygen

 

1.45

Microscopic Examination of Pure Cultures

 

1.46

The Use of Serology In Pure Culture Study

 

1.47

Maintenance and Preservation of Cultures

 

1.48

Selection of Materials

 

1.49

Cleaning Glassware’s

 

1.50

Preparation of Glassware For Sterilization

 

1.51

Sterilization

 

1.52

Titration of Media

 

1.53

Fuller's Scale

 

1.54

Platinum needles

 

1.55

Inoculating Tube Cultures

 

1.56

Caoutchouc Caps

 

1.57

Serum Inspissator

 

1.58

Old Apparatus for Storing and for Cultivations in Liquid Media

 

1.59

The Growth of the Microbial Cell in Pure Culture and in Mixed Populations

 

1.60

Principles of Microbial Nutrition and  Composition of Culture Media for Microorganisms

 

1.61

Cotton Filters

 

 

SECTION –I: CULTURE MEDIA

 

1

A1 Medium

 

2

A3 Agar

 

3

A7 Agar

 

4

ABY Agar (Acid Bismuth Yeast Agar)

 

5

Abeyta-Hunt Bark Agar

 

6

Actidione (Cycloheximide) Agar

 

7

Acetamide Agar

 

8

Acetamide Broth

 

9

Acetate Agar

 

10

Acetate differential

 

11

Acetivibrio celluloyticus Medium

 

12

Actinomyces Broth/Agar

 

13

Actinomycete Isolation Agar with Glycerol

 

14

Acetobacter diazotrophicus Agar

 

15

Acetobacterium dehalogenans Medium

 

16

Acetogen Medium

 

17

Acetohalobium Medium

 

18

Acetylglucosamine Medium

 

19

Acid Glucose Salts Medium

 

20

Acid Rhodospirillaceae Medium

 

21

Acid Egg Medium

 

22

Acid Tomato Broth

 

23

Acidic Tomato Medium for Leuconostoc

 

24

Actinomyces Broth

 

25

Actinomycete Growth Medium

 

26

Acidobacterium Medium

 

27

AE Sporulation Medium, Modified

 

28

Aeromonas Differential Agar

 

29

Aeromonas Medium

 

30

AFPA (Aspergillus flavus/parasiticus Agar)

 

31

Agrobacterium Agar

 

32

AK Agar 

 

33

AKI Medium

 

34

Alcaligenes Agar

 

35

Alginate Utilization Medium

 

36

Alkaline Cellulose Agar

 

37

Alkaline Nutrient Agar

 

38

Alkaline Peptone Agar

 

39

Alkaline Peptone Water

 

40

Alkaline Polypectate Agar

 

41

Alkaline Starch Agar

 

42

Alkaliphilic Methanogen Medium

 

43

Alkaliphilic Spirochete Medium

 

44

Alkalophile Medium

 

45

Alkalophilic Halophile Agar

 

46

Allantoin Agar

 

47

ALP Basal Medium

 

48

Amino-butyric Acid Medium

 

49

Amies Transport Medium With Charcoal

 

50

Amies Transport Medium Without Charcoal

 

51

Amino Acid Assay Media. Lysine Assay Medium • Cystine Assay Medium

 

52

AMS Agar

 

53

Ammonium Agar

 

54

Amygdalin Medium

 

55

Anaerobic Agar

 

56

Andrade’s Carbohydrate Broth and Indicator

 

57

Antibiotic Medium (Seed Agar)

 

58

Antibiotic Medium (Base Agar)

 

59

Antibiotic Medium

 

60

Antibiotic Medium (For Streptomycin Assays)

 

61

Antibiotic Medium (Base Agar With Low pH)

 

62

Anaerobic Agar

 

63

Anaerobic Cellulolytic Medium

 

64

AOAC Letheen Broth

 

65

AO Agar

 

66

Apple Juice Yeast Extract Medium

 

67

APT Agar/APT Broth

 

68

Arginine Broth

 

69

Armstrong Fusarium Medium

 

70

Artificial Deep Lake Medium

 

71

Artificial Organic Lake Peptone Medium

 

72

Arylsulfatase Agar

 

73

Ascospore Agar

 

74

Ashbey’s Nitrogen-Free Agar

 

75

Asparagine Broth

 

76

Asparagine Broth

 

77

Aureomycin Rose Bengal Glucose Peptone Agar

 

78

Autotrophic Nitrobacter Medium

 

79

Azide Blood Agar

 

80

Azide Blood Agar with Crystal Violet

 

81

Azide Broth

 

82

Azide Citrate Broth

 

83

Azospirillum Medium

 

84

Azotobacter Agar

 

85

Azotobacter Broth

 

86

Azotobacter chroococcum Agar

 

87

Azotobacter paspali Medium

 

88

Azotobacter vinelandii Medium

 

89

Baar’s Medium for Sulfate Reducers

 

90

Bacillus Agar

 

91

Bacillus halodenitrificans Agar

 

92

Bacillus Medium

 

93

Bacillus Medium

 

94

Selective Agar Base For Bacillus Cereus.

 

95

Bacillus thuringiensis Medium

 

96

BCP Azide Broth

 

97

Bdellovibrio Medium

 

98

Beef Extract Agar

 

99

Beef Extract Broth

 

100

Beggiatoa and Thiothrix Medium

 

101

Beijerinckia Medium

 

102

Beijerinck’s Thiobacillus Medium

 

103

Benzoate Medium

 

104

Bile Salts Gelatin Agar

 

105

Biphenyl Agar

 

106

Bismuth Sulfite Broth

 

107

Blood Agar

 

108

Blood Agar Base

 

109

Blood Agar With Nalidixic Acid Base

 

110

Blood Agar Base with 2.5% NaCl

 

111

Blood Glucose Cystine Agar

 

112

Blue-Green Nitrogen-Fixing Agar

 

113

Brackish Acetate

 

114

Brevibacterium Medium

 

115

Brilliant Green Bile Agar

 

116

Brilliant Green Bile Broth

 

117

Brilliant Green Bile Broth with MUG

 

118

Brilliant Green Broth

 

119

Bromcresol Purple Broth

 

120

Bromcresol Purple Dextrose Broth

 

121

Bromothymol Blue Agar

 

122

Bryant- Burkey Broth

 

123

Burke’s Modified Nitrogen-Free Medium

 

124

Brucella Broth

 

125

Bushnell-Haas Agar

 

126

Butanediol Medium

 

127

Camphor Minimal Medium

 

128

Carbohydrate Fermentation Broth

 

129

Carbon Monoxide Oxidizers Agar, Modified

 

130

Carbon Utilization Test

 

131

Carboxydobacterium Medium

 

132

Casein Medium

 

133

Caulobacter Medium

 

134

Cellulolytic Agar for Thermophiles

 

135

Cellulolytic Broth with Sea Salts

 

136

Cellulolytic Medium

 

137

Cellulolytic Medium with Rumen Fluid and Soluble Starch

 

138

Cellulomonas fermentans Medium

 

139

Cellulose Anaerobe Medium

 

140

Cellulose Broth

 

141

Cellvibrio Medium

 

142

Centenum Medium

 

143

Cetrimide Agar

 

144

Chitin Agar

 

145

Chitin Agar

 

146

Chlorobutane Medium

 

147

Chlorohydroxybenzoic Acid Medium

 

148

Cholera Medium TCBS

 

149

Citrate Medium, Koser’s Modified

 

150

Citrate Phosphate Buffered Glucose Medium

 

151

Clostridium beijerinckii Agar

 

152

Clostridium Cellulolytic Medium

 

153

Clostridium Cellulose Medium

 

154

Clostridium kluyveri Agar

 

155

Clostridium pfennigii Medium

 

156

Clostridium propionicum Medium

 

157

Clostridium thermohydrosulfuricum Medium

 

158

Colby and Zatman Agar

 

159

Colloidal Chitin Agar

 

160

Cook’s Cytophaga Agar

 

161

Cornmeal Agar(CMA)

 

162

Cornmeal Agar with Soil Extract

 

163

Cornmeal Yeast Extract Seawater Agar

 

164

Corynebacterium Medium with Salt

 

165

Cow Manure Agar

 

166

CP Medium for Coprothermobacter proteolyticus

 

167

Cyclohexanone Medium

 

168

Czapek Agar

 

169

Czapek Dox Agar, Modified

 

170

Czapek Dox Broth

 

171

Czapek Solution Agar with Sucrose

 

172

Deoxycholate Agar

 

173

Deoxycholate Citrate Agar

 

174

Deoxyribonuclease (DNase Test) Agar (pH 7.3)

 

175

Desulfovibrio Medium

 

176

Dextrose Broth

 

177

Dextrose Soil Agar(DSA)

 

178

Diazotrophic Medium(RBA)

 

179

Dibenzothiophene Mineral Medium

 

180

Dichloroacetic Acid Medium

 

181

Dichloromethane Medium for Hyphomicrobium

 

182

DNase Medium

 

183

Dubos Agar with Filter Paper

 

184

Dubos Mineral Medium

 

185

Dubos Salts Agar

 

186

Dubos Salts Broth with Yeast Extract

 

187

E Agar(m-E Agar)

 

188

EC Broth(Escherichia coli Broth)(EC Medium)

 

189

EC Broth with MUG

 

190

EIA Substrate

 

191

Eijkman Lactose Medium

 

192

EMB Agar(Eosin Methylene Blue Agar)

 

193

EMB Agar, Modified(Eosin Methylene Blue Agar, Modified)

 

194

Emerson Agar

 

195

Endo Agar

 

196

Endo Agar, LES(Endo Agar, Laurance Experimental Station) (m-Endo Agar, LES)(m-LES, Endo Agar)

 

197

Endo Broth(m-Endo Broth)

 

198

Enriched Cytophaga Agar

 

199

Enterococci Confirmatory Agar

 

200

Enterococci Presumptive Broth

 

201

Enterococcus Agar(m-Enterococcus Agar)(Azide Agar)

 

202

Erwinia Fermentation Medium

 

203

Esculin Agar

 

204

Esculin Iron Agar

 

205

Ethyl Violet Azide Broth(EVA Broth)

 

206

Ethyl Violet Azide Broth(EVA Broth)

 

207

Ethyl Violet Azide Broth(EVA Broth)

 

208

Extracted Hay Medium

 

209

EYGA Agar

 

210

Fay and Barry Medium

 

211

FC Agar (Fecal Coliform Agar)(m-FC Agar) (m-Fecal Coliform Agar)

 

212

Fecal Coliform Agar, Modified(m-Fecal Coliform Agar, Modified)(FCIC)

 

213

Fecal Coliform Agar, Modified

 

214

Feodorov Medium

 

215

FGTC Agar

 

216

Fish Peptone Broth

 

217

Flavobacterium Medium

 

218

Fletcher Medium

 

219

Flo Agar

 

220

Fluorescent Pectolytic Agar(FPA Medium)

 

221

FN Medium(Fluorescence Denitrification Medium)

 

222

FPA Medium

 

223

Frankia Agar

 

224

Fraser Broth

 

225

Freshwater Ameba Medium

 

226

FSM Selective Medium

 

227

FUF Medium

 

228

Gelatin Agar

 

229

General Salts Medium for Estuarine Methanogens

 

230

Geo Medium

 

231

George’s Medium, Modified

 

232

Gisa Agar

 

233

Gluconate Peptone Broth

 

234

Glucose Agar, 9K

 

235

Glucose Agar with 25%Glucose

 

236

Glucose Nitrogen-Free Salt Agar

 

237

Glucose Peptone Agar

 

238

Glucose Yeast Chalk Agar

 

239

Glucose Yeast Extract Agar

 

240

Glutamate Medium

 

241

Glycerol Agar

 

242

Glycerol Arginine Agar

 

243

Glycerol Chalk Agar

 

244

Gorham’s Medium for Algae

 

245

Hagedorn and Holt Selective Medium

 

246

Haloalkaliphilic Agar

 

247

Halobacteriaceae Medium

 

248

Halobacterium halobium Defined Medium

 

249

Halophile Agar

 

250

Halophile Medium

 

251

Halophilic Broth(HB)

 

252

Halophilic Nutrient Agar

 

253

Halophilic Synthetic Medium

 

254

Hartley’s Digest Broth

 

255

Hay Infusion Agar

 

256

HE Medium(Hay Extract Medium)

 

257

Heart Infusion Agar

 

258

Hektoen Enteric Agar

 

259

Heterotrophic Hyperthermophilic Archaea Medium

 

260

Heterotrophic Medium H3P

 

261

Heterotrophic Medium for Hydrogenomonas

 

262

Heterotrophic Medium for Hydrogen-Oxidizing Bacteria

 

263

Heterotrophic Nitrobacter Medium

 

264

HPC Agar (Heterotrophic Plate Count Agar) (m-HPC Agar)

 

265

Hugh-Leifson’s Glucose Broth

 

266

Modified Hutner’s Basal Salts

 

267

Hydrogen-Oxidizing Bacteria Medium

 

268

Hydroxybenzoate Agar(p-Hydroxybenzoate Agar)

 

269

Hyphomicrobium Enrichment Medium

 

270

Imhoff’s Medium, Modified

 

271

Imidazole Utilization Medium

 

272

Inorganic Salts-Maltose Medium

 

273

Inositol Brilliant Green Bile Salts Agar(IBB Agar)

 

274

Iron Bacteria Isolation Medium

 

275

Iron-Oxidizing Medium

 

276

Iron Sulfite Agar

 

277

Isoleucine Hydroxamate Medium

 

278

Johnson’s Marine Medium

 

279

Kerosene Mineral Salts Medium

 

280

Ketolactonate Broth

 

281

Kligler Iron Agar (pH 7.4)

 

282

Kligler Iron Agar

 

283

King’s Medium

 

284

Kleb Agar(m-Kleb Agar)

 

285

Koser Citrate Medium

 

286

KYE Agar

 

287

Lactate Agar

 

288

Lactose Broth

 

289

Lactose Minimal Medium

 

290

Lactose Ricinoleate Broth

 

291

Lauryl Sulfate Broth(m-Lauryl Sulfate Broth)

 

292

Lauryl Sulfate Broth(Lauryl Tryptose Broth)

 

293

Lauryl Sulfate Broth with MUG

 

294

Lauryl Tryptose Mannitol Broth with Tryptophan

 

295

LB Agar

 

296

Lead Acetate Agar

 

297

Legume Extract Agar

 

298

Leptospira Medium

 

299

Leuconostoc Medium

 

300

Leucothrix mucor Medium

 

301

Levine EMB Agar(Levine Eosin Methylene Blue Agar) (Eosin Methylene Blue Agar, Levine)(LEMB Agar)

 

302

Lima Bean Agar

 

303

Lipovitellin Salt Mannitol Agar

 

304

Litmus Milk

 

305

Lysine Decarboxylase Broth, Falkow

 

306

Lysine Decarboxylase Broth, Taylor Modification

 

307

Lysine Decarboxylase Medium

 

308

Lysine Iron Agar

 

309

MacConkey Agar

 

310

MacConkey Broth

 

311

MacConkey Broth, Purple

 

312

Malachite Green Broth

 

313

Malonate Broth

 

314

Malt Agar

 

315

Manganese Acetate Agar

 

316

Manganese Medium for Pseudomonas species

 

317

Mannitol Yeast Extract Medium

 

318

Marine Agar

 

319

Marine Agar with Biphenyl

 

320

Marine Agar with Naphthalene

 

321

Marine Flagellate Medium

 

322

Marine Rhodococcus Medium

 

323

Marine Rhodopseudomonas Medium

 

324

Medium for Ammonia Oxidizers, Marine

 

325

Medium for Carbon Monoxide Oxidizers

 

326

Medium F for Sulfate Reducers(Postgate’s Medium F for Sulfate Reducers)

 

327

Medium for Halophilic Bacilli

 

328

Medium for Hydrocarbon-Degrading Bacteria

 

329

Medium for Hydrocarbon-Degrading Bacteria (Naphthalene Mineral Salts Medium)

 

330

Medium for Nitrite Oxidizers

 

331

Medium for Nitrite Oxidizers, Marine

 

332

Medium for Osmophilic Fungi

 

333

Medium for Thermophilic Actinomycetes

 

334

Membrane Lauryl Sulfate Broth

 

335

Methanobacillus Medium

 

336

Methanobacteria Medium

 

337

Methanobacterium thermoautotrophicum MS Medium

 

338

Methanogen Enrichment Medium, Barker

 

339

Methanogen Medium

 

340

Methanol Agar

 

341

Methylamine Salts Medium

 

342

Mineral Medium with 2,4- Dichlorophenoxy acetic Acid

 

343

Mineral Medium, Nagel and Andreesen

 

344

Mineral Medium with Naphthalene

 

345

Mineral Medium with Phenol

 

346

Mineral Medium with Pyrrolic Acid

 

347

Mineral Medium with Salicylate

 

348

Mineral Medium with Toluene

 

349

Moller Decarboxylase Broth

 

350

Moller KCN Broth Base

 

351

Motility Test Media (pH 7.2)

 

352

MPH Agar(Milk Protein Hydrolysate Agar)

 

353

MPSS Broth

 

354

MRVP Broth(Methyl Red Voges-Proskauer Broth)

 

355

Mycorrhiza Medium

 

356

Myxobacteria Medium

 

357

Nine K Medium (9K Medium)

 

358

Nitrate Agar

 

359

Nitrate Reduction Broth for Pseudomonas and Related Genera

 

360

Nitrogen-Fixing Hydrocarbon Oxidizers Medium

 

361

Nitrogen-Fixing Marine Medium

 

362

Nitrogen-Free Agar

 

363

Nitrogen-Free Agar (Norris Agar)

 

364

Nitrosomonas Medium

 

365

Nutrient Agar

 

366

Nutrient Agar pH 10.0

 

367

Nutrient Soil Extract Agar

 

368

NWRI Agar(HPC Agar)

 

369

Oat Flake Medium

 

370

Oil Agar Medium

 

371

Oxidation-Fermentation Medium(OF Medium)

 

372

Oxidation-Fermentation, Medium, King’s (King’s OF Medium)

 

373

PA-C Agar(mPA-C Agar)

 

374

Paraffin Agar

 

375

Payne, Seghal, and Gibbons Medium