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216 Manual of basic techniques for a health laboratory5.9 Examination of watery stool specimensDark-field microscopy is used to identify Vibrio cholerae and Campylobacter spp. inwatery stool specimens.5.9.1 Materials and reagents●●●●Microscope with dark-field attachmentMicroscope slidesCoverslipsInoculating loop● Sodium chloride, 0.85% solution (reagent no. 53).5.9.2 Method1. Suspend 0.2g of stool in 5ml of sodium chloride solution. Allow the large particlesto sediment.2. Using an inoculating loop (sterilized by flaming), prepare a very thin smear on aslide. Carefully remove any large particles.3. Cover with a coverslip. Place the slide on the microscope stage.4. Open the iris diaphragm fully and place the dark-field attachment in position.5.9.3 Microscopic examinationUse the ¥ 10 objective for focusing. The background appears black, and all objectssuspended in the saline solution appear bright.Use the ¥ 40 objective to search for bacteria with characteristic shapes and motility(see below).Vibrio cholerae appears as motile rods, which may be short, curved, straight or involuted(Fig. 5.34).Campylobacter spp. are Gram-negative spiral rods that rotate rapidly on a centralaxis.5.9.4 Dispatch of specimens for culture 1It is often necessary to send stool specimens to a bacteriology laboratory forculture:— for the detection of cholera vibrios— for the detection of other bacteria that cause dysentery (species of Salmonella,Shigella, etc.).Using Cary–Blair transport mediumCary–Blair transport medium (reagent no. 17) will preserve many kinds of entericbacteria (cholera vibrios, other vibrios, salmonella, shigella, etc.) for up to 4 weeks.The uninoculated medium may be stored in a sealed bottle at room temperaturefor 8–12 weeks.1. Dip a sterile cotton wool swab in the stool specimen (Fig. 5.35).2. For infants or other patients who cannot produce a stool specimen, take a rectalswab. Moisten the swab with sodium chloride solution and introduce the swabinto the rectum. Turn the swab several times with a circular movement (Fig.5.36).1See also section 3.7.1.
5. Bacteriology 217Fig. 5.34 Vibrio cholerae3. Place the swab in a bottle containing Cary–Blair medium (three-quarters full)and send it to the bacteriology laboratory. If you cannot send the swab immediately,store it at room temperature.Important:●●Never store the swab in the incubator.Never store the swab in the refrigerator.Using buffered glycerol salineWhen specimens are to be sent for culture of enteric organisms other than choleravibrios and Cary–Blair transport medium is not available, buffered glycerol saline(reagent no. 14) may be used.Note: If the buffered glycerol saline has changed colour from pink to yellow, discardit and prepare a fresh solution.Fig. 5.35 Collecting a waterystool specimenFig. 5.36 Collecting a stool specimen from an infant
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M A N U A LO F B A S I CTECHNIQUESF
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ContentsiManualofbasic techniques f
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ContentsiiiContentsPrefacex1. Intro
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viiiManual of basic techniques for
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Table 3.2 Dispatch of specimens 92
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Page 243 and 244: 7. Examination of urine 231Part III
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Page 249 and 250: 7. Examination of urine 237Table 7.
Page 251 and 252: 7. Examination of urine 239The anal
Page 253 and 254: 7. Examination of urine 241MethodCo
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Page 257 and 258: 7. Examination of urine 245Fig. 7.2
Page 261 and 262: 7. Examination of urine 2497.2.8 De
Page 263 and 264: 7. Examination of urine 251Reuse of
Page 267 and 268: 8. Examination of cerebrospinal flu
Page 277 and 278: 9. Haematology 2659. HaematologyHae
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9. Haematology 2679.2 Collection of
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9. Haematology 269Fig. 9.13 Feeling
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9. Haematology 2719.3 Estimation of
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9. Haematology 273Table 9.2 Sample
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9. Haematology 275Fig. 9.25 Checkin
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9. Haematology 277Calibration of th
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9. Haematology 279Errors in haemogl
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9. Haematology 281Fig. 9.28 Micro s
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9. Haematology 283Fig. 9.34 Centrif
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9. Haematology 285concentration is
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9. Haematology 287Measurement techn
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9. Haematology 289●●●Graduate
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9. Haematology 2911 litre, so multi
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9. Haematology 293Fig. 9.45 Materia
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9. Haematology 295Very high ESR val
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9. Haematology 2979.9 Observation o
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9. Haematology 2999.10 Preparation
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9. Haematology 301Fig. 9.64 Making
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9. Haematology 303Drying the filmAd
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9. Haematology 3054. Examine the co
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9. Haematology 307Fig. 9.77 Normal
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9. Haematology 309Fig. 9.85 Anisocy
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9. Haematology 311Fig. 9.92 Polymor
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9. Haematology 313Immature granuloc
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9. Haematology 315Fig. 9.104 Mixing
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9. Haematology 317Fig. 9.108 Prepar
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9. Haematology 319Heinz bodiesHeinz
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9. Haematology 3219.14 Determinatio
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10. Blood chemistry 323Fig. 10.1 De
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10. Blood chemistry 325Table 10.1 B
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10. Blood chemistry 32710.2.4 Resul
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11. Immunological and serological t
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Annex. Reagents and their preparati
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IndexNote: Page numbers in bold ref
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Index 371Centrifuges 32, 69-72, 69-
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Index 373Endolimax nanus 114, 114,
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Index 375Hyaline casts, urine 243,
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Index 377Microhaematocrit equipment
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Index 379Protozoablood 173-194, 178
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Index 381Smears (continued)preparat
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Index 383Thiomersal-iodine-formalde
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