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III. METHODS,

1. Microscopical Investigation of Micro-Organisms . .

Preparation Making . . . . . .

Removal of Grease from Cover Glasses . .

Fixing and Staining of Yeast Cells

Staining of Yeast Spores . . . .

Staining of the Yeast Cell Nucleus .

Fixing and Staining of Bacteria . . .

Staining of Bacteria Spores. . .

Staining of Flagella according to Löffler .

Reagents . . . . . . . .

Development in Moist Chambers . . .

2. Experiments with various Flasks. Inoculation of Liquid and

Solid Culture Media . . . . . . .

The Manipulation of Nutrient Liquids .

Experiments with Solid Culture Media .

Cultures of Anaërobic Organisms . . .

Suppression of Bacteria in Yeast Growths.

3. Preparation of Pure Cultures . . . .

Brefeld's Glass Slip Cultures

Hansen's Dilution Method

101

Dilution in and on Solid Substrata

103

Koch's Plate Culture . .

103

Surface Plate Cultures.

Hansen's Second Pure Culture Method .

. 106

Lindner's Droplet Culture . . . .

110

Schönfeld's Method . .

111

Fractionated Culture after Klebs and others

111

Pasteur's Method . . .

112

Pure Cultures of Bacteria . . . . . . . 113

Pure Cultures of Mould Fungi . .

. . 113

4. Methods of Preservation

. . 114

Hansen's Saccharose Method for the Preservation of Yeasts

and Moulds . . . . . . . . . 114

Preservation on Cotton Wool or Filter Paper (after Hansen) 117

Preservation of Bacteria

117

Preservation of Ordinary Brewery Yeast . . . . 118

Transmission of Yeast . . . . . . . . 119

5. Preparation of Spore Cultures .

121

Spore Cultures of Saccharomyces . .

121

Spore Cultures of Bacteria . . . . . . . 124

Spore Cultures of Moulds .

124

6. Preparation of Film Cultures of Saccharomyces . . . . 125

7. Counting of Yeast Cells and Seeding with a Definite Number

of Cells . . . . . . . . . . . 126

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137

138

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2. THE HIGHER FUNGI (MYCOMYCETES) . . . . 186

A. ASCOMYCETES . . . . . . .

. 186

1st ORDER : GYMNOASCEÆ . . . . .

186

True Yeast Fungi (Saccharomycetes) . .

General

186

1. The Saccharomycetes Distinct Fungi. . 186

2. Structure and Shape of Yeast Cells . .

The Cell Contents . . . . .

187

The Cell Wall and its Gelatinous Formation 189

Shape of the Cells . .

3. Modes of Propagation . . . . .

191

Budding

191

Sjore Formation . . . . . . .

209

4. The Chemical Constituents of the Cell. . . 211

5. Fermentation Phenomena . . . . ..

212

Ferments . . . . . .

Influence of Air and Temperature on Fermenta-

tion . . . . . . . . . 214

Energy of Fermentation and Fermenting

Power . . . . . . . . . 217

Permeability of the Cell Membrane . . . 217

Products of Fermentation . . . . . 218

Auto-Fermentation . . . . . . 219

Yeast Types . . . . . . . .

6. Injurious and Stimulating Influence of Chemical

and Physical Factors . . . . . 222

Influence of Chemical Factors . . . . 222

Influence of Physical Factors . .

224

7. Vitality of Yeast in Nutrient Solutions and in

the Dry State . . . . . . . . 227

8. Disease Yeasts and Mixed Fermentations . .

Diseases caused by Fermentation Products

Diseases caused by Turbidity

. 229

Competitive Relations . . . . . 229

9. Variation . . . . . . . 233

Temporary Varieties . .

Permanent Varieties . .

Practical Results and Variation in Practice

10. Circulation in Nature . . . . . . . 246

Systematic . . . . . . . . .

1. Saccharomyces . . . . . . . 249

General Characteristics, p. 249; Grouping according

to Action on Sugars, p. 249 ; S. cerevisiæ I., p. 251 ;

Carlsberg Bottom Yeast No. 1, p. 252; Carlsberg

Bottom Yeast No. 2, p. 252; Four Species of Brewery

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Bottom Yeast Described by Will, p. 252; Beer Top

Yeast, p. 253; Distillery Top Yeast, p. 253; S. Pas-

torianus I., p. 254; S. Pastorianus II., p. 255 ; S.

Pastorianus III., p. 256; S. ellipsoideus I., p. 257 ;

Johannisberg II., p. 258 ; Walporzheim, p. 258 ;

S. ellipsoideus II., p. 258; Two Disease Yeasts

Isolated by Will, p. 259 ; S. Ilicis, p. 259; S. Aqui-

folii, p. 260; S. Vordermanni, p. 260; S. pyriformis,

p. 261 ; S. Marxianus, p. 261 ; S. exiguus, p. 262;

S. anomalus, p. 262; S. membranæfaciens, p. 264 ;

S. Bailii, p. 265 ; S. mali Duclauxi, p. 265; S. car-

tilaginosus, p. 266; $. fragilis, p. 266 ; Kefir, p. 266;

Mazun, p. 267 ; S. Ludwigii, p. 267.

2. Schizosaccharomyces . . . . . . 269

Sch. Pombe, p. 269 ; Sch. octosporus, p. 270 ; Sch.

mellacei, p. 271.

2ND ORDER : PERISPORACEÆ . . . . . . . 271

General Characteristics, p. 271; Anixiopsis stercoraria,

p. 272; The Relation between Vegetable Growth

and Perithecium Formation, p. 272; Damage in the

Brewery, p. 273; Preservation in the Laboratory,

p. 273.

Aspergilleæ . . . . . . . . . . 274

1. Aspergillus . . . . . . . . . 274

A. glaucus, p. 274; A. oryzæ, p. 277 ; Manufacture of

Saké, p. 277; A. fumigatus, p. 277; A. niger, p.

277.

2. Penicillium . . . . . . . . . 278

P. glaucum, p. 278.

3RD ORDER: SPHÆRIACEÆ . . . . . . . 282

Sphæriex . . . . . . . . . 282

1. Sphærella . .

. . . . . 282

Sph. Tulasnei, p. 283; The Conidial Form Clados-

porium herbarum, p. 283 ; Hormodendron clados-

porioides, p. 285.

4TH ORDER: DISCOMYCETES . . . . . . 285

Cup Fungi, Pezizaceæ . . . . . . 286

Sclerotinia ·

. . 286

S. Fuckeliana, p. 286 ; The Conidial Form Botrytis

cinerea, p. 286.

B. Fungi IMPERFECTI . . . . . . . . 289

The Torula Species . .

. . 289

Hansen's Seven Torula Species, p. 290; Rich. Meiss-

ner's Eleven Slime Yeasts, p. 292.

Saccharomyces apiculatus . . . . . . 294

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