district laboratory practice in tropical countries, color plates

8/10/2019 District Laboratory Practice in Tropical Countries, Color Plates

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COLOUR SECTION

1 Indole and ureasetests. Right: A positiveindole strip tesl. Left: Aposttlve urease tesl.

6 Right : Methylene blue stained pus cells in faecesas seen with 40x objective. A mononuclear cell can beseen upper left. Left: The unstained yellow cells arered cells. see7.18.11.

8 Yellow, sucrose fermenting Vibriochoterae colonies on tnlosutphatecitrate bile-salt sucrose (TCBs) agar.

2 Indole test (tryptone water culture) Right: A positive test. Left: Anegative lest. See 7.5.6

4 Urease test (Christensen'surea broth). Right: A positive lestLeft: A neqatlve test. See 7.5.9

5 Lilmus milk decolor izat iontest, Right: A posttlve test. Left: Anegative lest. See 7.5.7

7 Gram stained smear (using 1 in 10 carbol fuchsin as coun-terstain) of alkaline peptone water culture of faeces showingVibrio cholerae, as seen wilh the 100x objective. See 7.18.19

9 Mixed culture (36 h) o f Vibriocholerae and enterococci (very smallcolonies) on TeBS agar.

Plates 1-10

10 Blue-green, non-sucrose ferment-

ing colonies of Vibrio parahaemolyticuson TCBS agar.


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DISTRICT LABORATORY PRACTICE IN TROPICAL COUNTRIES

11 Xylose lysine deoxycholate (XLD) cultures. Left: YellowEscherichia coli colonies. Right: Red-pink Salmonellacolonies (some Proteus species look identical). See 7.11.

13 Kligler iron agar cultures of salmonellae and shigellae showinga pink-red slope (alkaline reaction) and yellow butt (acid reaction). AShigella species. B: S. Paratyphi A, note the cracks in the agar dueto gas product ion . C: $. Typhi, note the small amount of blackeningdue to H 2S production. , ? : S . Typhimurium, ro te the break up of theagar due 10 gas production and large amount of H 2S produced

14 Deoxycholate citrate agar (DCA} cultures. Left: Non-lactose fermenting Shigella colonies. Right" Lactosefermenting Escherichia coli colonies. See 7.11.

15 Basic fuchsin stained Gampy-

/abacter species as seen wi th the 100Xoil object ive . Note, smal l spi ra l curved'gull ' forms and older spirochaetal forms

16 Campy/obacter jejuni on Butzler selec-tive medium cultured at 37C in a candle jar.See 7.18.21.

17 Close-up of typical droplel-like spreading colonies of Campy-lobacterjejuni. See 7.18.21.

Plates 11- 17


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COLOUR SECTION

18 Urine cultures on CLED agar. Upper left: Enterococcusfaecalis. Upper right: Proteus species. Lower left :Escherichia coli. Lower righf: Klebsiella species. See 7.12.

20 Wagtech portable water testing kit being used in thefield. The operator is shown filtering the water through themembrane. Note the turbidity tubes on the right. See 7.17

22 Blood agar culture showing Staphylococcus aureus

(large colonies) and small beta-haemolytic colonies of Streptococcus pyogenes. See 7.18.1, 7.18.2.

19 Urine cultures on CLED agar using filter paper inocu-lation technique. Left, top downwards: E. faecalis, mixedgrowth, no significant growth, no growth. Right, top down-wards: Klebsella species, Proteus species, Staphylococcusspecies, E. coli

21 Membrane culture of water sample after incubation at44C for 24 h. There are approx. 40 colonies on themembrane, corresponding to a faecal coliform count per 100 ml of 800 (5 ml of water filtered). See 7.17

23 Pure growth of Staphylococcus aureus on blood agar.See 7.18.1.

Plates 18-23


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24 Gram positive cocci of Staphylococcus aureus insmear of pus, as seen using the 100X oil objec tive. See7.18.1.

26 Blood agar cultures. Upper. Culture of Streptococcus pyogenes showing sensi tivi ty to baci-tracin (no haemolysis around the disc). Lower.Beta-haemolytic bacitracin resistant streptococci (typedas Group F). See 7.18.2.

28 Streptococcus pneumoniae in Gram stainedsmear. The organisms are diplococci and surrounded bya capsule (seen as a p ink area around the organisms).8ee7.18.4.

25 Gram positive Streptococcus pyogenes in smear of pus, as seen using the 100x oil objective. See 7.18.2.

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27 Throat swab culture showing beta-haemolyticStreptococcus pyogenes. See 7.18.2.

29 Culture of Streptococcus pneumoniae on blood agar showing alpha-haemolytic colonies sens itive to optochindisc. See 7.18.4.

Plates 24-29


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COLOUR SECTION

30 Corynebacterium diphtheriae growing on telluriteblood agar. See 7.18.7.

32 Albert stained smear of Corynebacterium diphthe-riae rods showing dark-staining volutin granules as seenwith the 100x objective. See 7.18.7.

34 Clostridium perlringens in Gram stained smear asseen with the 100X objective. See 7.18.9.

31 Corynebacterium diphtheriae cultured on modifiedTinsdale medium. Colonies are surrounded by zones of brown discoloration. See 7.18.7.

33 Gram positive (easily decolorized) pleomorphic rods of Corynebacterium diphtheriae joined together at variousangles, as seen with the 100x objective. see 7.18.7.

35 Lactose egg-yolk milk agar cultures with antitoxinapplied to left half of plate. Upper. Clostridium perfingensshowing opacity due to lecithinase activity (Inhibited on leftby antitoxin) and reddening in medium due to lactose fer-mentation. Lower. Clostridium species showing lecithinaseactivity (partially inhibited) and clearing in medium due toproteolysis. See 7.18.9

Plates 30-35


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36 Clostridium perfringens showing marked haemoly-sis on blood agar after anaerobic incubation. See 7.18.9

38 Gram stained smear of cerebrospinal Iluid showingpus cells and Neisseria meningitidis (small intracellular Gram negative diplococci). See 7.18.12

4() Cervical swab culture of Neisseria gonorrhoeae(smallest colonies) on chocolate agar. Because themedium is non-selective there is no inhibition of vaginalflora. See 7.18.13.

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37 Gram stained smear of culture of Clostridium fetanishowing terminal spores

39 Neisseria meningitidis colonies on chocolate agar after overnight incubation in a carbon dioxide enrichedatmosphere. See 7.18.12.

41 Pure growth of Neisseria gonorrhoeae on Thayer Martin agar, made selective by the addition of antibiotics.See 7.18.13.

Plates 36-41


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COLOUR SECTION

42 Pure growth of Neisseria gonorrhoeae on modifiedNew York City selective medium. See 7.18.13.

44 Acridine orange fluorescent stained vaginal smear showing Tn'chomonas vagina/is (red-brown) and pus cells(yellow-green), as seen with the 40X objective. See 7.10.

46 Romanowsky stained urethral smear showingKlebsiella granulomafis (Donovan bodies) in amacrophage cell as seen with the 100x objective. Theorganisms show bi-polar staining. See 7.10

43 Gram stained urethral smear showing pus cells andNeisseria gonorrl1oeae (intracellular Gram negative diplo-cocci ) as seen with the 100X object ive. See 7.18.13. Note:

~~!a.~~On1Joeae looks the same as N. meningitidis in

45 Clue cell in Gram stained vaginal smear showing

Gram negative bacilli and Gram variable coccobacilli asseen with the 100X objective. See 7.10

47 Pseumomonas aeruginosa on blood agar. Note

pigment in the medium. See 7.18.20

Plates 42-47


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"48 Gram negative pleomorphic rods of HaemophiJus

influenzas in cerebrospinal fluid, as seen with the 100X oilobjective. Note the thread-like forms. See 7.18.24.

52 Positive sateutnsm test to identify Haemophilusinfluenzae. The Haemophilus colonies are growing onlynear the central Staphylococcus aureus line 01 growthSee 7.18.24.

49 Haemophilus influenzae colonies on chocolate agar af ter overnight incubat ion in a carbon dioxide enr ichedatmosphere. See 7.18.24.

50 Beta-Iaclamase acidimetric filler paper test. Right:Yel low react ion of oera-tactamase producing strain. Left: Nocolour change of a non-beta-Iactamase producing strain. See7.16.

51 Oxoid nitrocefin test to detect beta-Iactamase produc-. . ing strains. Left: Strongly positive test. Centre: Weakly

positive test. Right: Negative eera-tactamass strain. See

53 Identi ficat ion of H. influenzae using X, V, XV discsGrowth is seen around XV disc. See 7.18.24.

Plates 48-53


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COLOUR SECTION

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54 Yersinia pestis in Giemsa s ta ined smear as seen wi th thelOOX objective. The organism shows bi-polar staining. See7.18.22.

56 Mycobacterium tuberculosis acid fast bacilli (AFB) inZlehl-Neelsen stained sputum smears counterstained withmalachite green. Left: Many AFB. Right: Scanty AFB (100xobjective). See 7.18.28

58 Mycobacterium leproe in Ziehl-Neelsen stained skinsmear as seen with the lOOx objective. See 7.18.29.

Plates 54-59

55 Loeffler polychrome methylene blue stainedblood smear showing anthrax bacilli surrounded bymauve-red capsules (McFadyean's reaction), as seenwith the lOOX oil objective. See 7.18.6.

57 Many Mycobactenum tuberculosis acid tastbacilli (AFB) in Ziebt-Neelsen stained sputum smear couoterstaoeo with methylene blue (100x objective).See 7.18.28.

J59 Gram positive branching intertwining threads of

actinomycetes as seen with the 100x objective. See7.18.31.


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60 Treponema pallidum in chancre fluid as seen by dark-field microscopy using the 40X objective. See 7.18.32.

'fIi,H,e'Syphilis

'fIi,H ,cf Syphilis

Control~

Test area

Samplewell

Positive test Negative test

62 Visitect Syphilis test results. See 7.18.32

64 Borreliae in Romanowsky stained blood films frompatient with relapsing fever as seen with 1OOx objective. Left:Bcrreliae in thin blood film (Giemsastained). Right: Borreliaein thick film (Field stained). See 7.18.34.

61 Leptospira interrogans as seen by dark-fieldmicroscopy using the 40x objective. See 7.18.33.

Negative Positive

63 LeptoTek Ori-Dot latex agglutination test to detectspecific Leptospira antibodies. See 7.18.33

65 Borrelia vincenti and Gram negative anaerobes inGram stained smear from a patient with acute ulcerativegingivitis. as seen with 100x objective. See 7.18.34.

Plates 60-65


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COLOUR SECTION

66 Giemsa stained blood film showingBartonella bacil/iformis organisms insidered cells from a patient with Oroya fever.See 7.18.36

67 Left: Giernsa stained smear showing Chlamydia trachoma tisinclusion bodies in epi the lial cel l, as seen with 100x object ive . Thenucleus of the ce ll (cent re) i s surrounded by b lue-mauve re ticulatebodies (above and to lower right of nucleus). Surrounding the epithelialcell are pus cells. Right: Chiamydia trachoma tis in an iodine preparationof conjunctival scraping. The inclusion body stains brown. See 7.18.37

68 Potassium hydroxide preparation of a s kin scale, showingringworm fungi with branching septate hyphae and spherical arthro-conidia. See 7.18.38.

70 Aspergillus in Gram stained sputum smear showing Gram

positive hyphae as seen with the l00x objective. See 7.18.49

Plates 66-71

69 Blue ink stained potassium hydroxide prep-aration showing Malassezia furfur with yeast cellsand short hyphae as seen with the 40X objective.5007.16.39

71 Intracellular yeast celis of Peniciiliummameffei in a Giemsa stained preparation asseen with the 100x objective. See 7.16.50.


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D,STRla LABORATORY PRACTICE IN TROPICAL COUNTRIES

72 Gram stained vaginal smear showing Gram posi tiveCandida albicans yeast cells and pseudohyphae as seenwith 100x objective. See 7.18.47

74 Yeast cells of Cryptococcus neoformans in Gramstained c.s.f as seen with the tuox object ive . See 7 .18.48.

76 Toluidine blue 0 (TBO) s ta ined smear showing spher-ical cysts of Pneumocystis jiroveci, as seen with the 40xobjective. T80 does not stain the internal structure of the

cysts. See 7.18.52.

73 India ink (Pelikan drawing ink) preparation showings ingle and budding yeast cel ls of Cryptococcus reotomansin c.s.! as seen with the 40x objective. Note the largecapsule surrounding the yeast cells. See 7.18.48.

75 Encapsulated Cryptococcus neoformans in Giemsastained c.s.t as seen with the 100X object ive. See 7 .18.48

77 Giemsa stained smear showing small internal struc-tures of Pneumocystis jiroveci cys ts (centre) as seen wi ththe 100x objec tive. Giemsa does not s tain the outl ine of thecysts. See 7.18.52.

Plates 72 77


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COLOUR SECTION

Positive reaction using serum

Negative reaction using serum

Positive reaction using whole blood.

Negative reaction using whole blood . . .. .-

78 Capillus HIV-1/HIV-2 assay to detectantibody to HIV-1/-2. See 7.18.55.

___ Control

----Tcstlinc

Posttlve Htvtest

Negative Hrvtcsr

79 UnlGold HIV 1 and 2 tes t results . See 7.18.55

HIVTESTS

I HIV 1I

\~ Positive

/ \ test

Control HIV-I Sample

line well

HIY-112HlV2

II Positive

/ \ \ test

Control HIV-2 Sample

line well

HIY-112 r: HIV 1, 2

I Negative

/ L

\ test

Control Samplewcll

80 SO Bioline HIV U 3.0 test results. See 7.18.55

Plates 78-80


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81 Normal blood film showing from left to right a monocyte, large lympho-

cyte, and neutrophil. See 8.7.

83 Normal blood film showing from left to right, a small lymphocyte,monocyte, and two neutrophils. See 8.7.

INFECTION BLOOD PICTURE

65 Infection blood picture showing left shift of neutrophils withband cells and metamyelocytes. See 8.7.

Plates 81-86

64 Basophil in a normal blood film(not often seen).

86 Blood film from a patientwith infection showing left shift of neutrophils with toxic granulation.


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COLOUR SECTION

87 BI?Od film from a patient with chronic lymphocyticleukaemia. Note the smear cells, l .e. damaged lymphocytes.

89 Blood film from a patient with chronic myeloid leukaemiashowing early myeloid cells. See 8.2.

91 Blast cell showing clear nucleoli. See 8.2

Plates 87-92

88 Blood film from a patient with acute lymphoblas-tic leukaemia. The cells are blast cells. See 8.2.

90 Myeloblasts showing Auer rods in cytoplasmand a lso in nucleus. See 8 .2 .

92 Imprint (touch) smear showing Burkitt'slymphoma cells. See 8,2.


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93 Blood film from patient with 94 Normal l arge lymphocyte, lower lightmyelomatosis showing a plasma cell, and reactive (atypical) lymphocytes to leftrouleaux of red cells and some back- See 8.7.ground protein staining. See 8.2.

95 Copack Haemoglobin Colour Scale showing haemoglobin value closeto 6g1dl. See 8.4.

96 Normocytic normochromic red cells.

REO CELLS

97 Blood film from a patient with macrocytic anaemia (folatedeficiency), showing oval macrocvtes. poikilocytosis, megaloblast

{right) and hypersegmented neutrophil. see 8.2, 8.4.

Plates 93-97


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COLOUR SECTION

98 Hypochromic microcytic red ce ll s f rom a pat ientwith iron deficiency. Note, elongated pencil cells. See8.2,8.4.

100 Positive sickle cell slide test. See 8.10. The sicklecells may appear crescent shaped with pointed ends or holly leaf shaped, especially in s ickle cell frait. Aposi tive sickle ce ll test indica tes tha t a person's redcells contain HbS.

102 Thick f ilm f rom a patient wifh s ickle cel l anaemiaand talclparum malar ia . Note the b lue s tippl ing in thebackground (reticulin of renculocytes). P . falciparumtrophozoite is shown in upper right of picture. See 8.1 O.

99 Blood f ilm f rom a child with sickle ce ll anaemia, showingsickle cells, nucleated red cell, tarqet cells and polychromasia.See8.1D.

101 HbS solubility filtration test. Left: Result with normal adulthaemoglobin (HbAA) showing dark red filtrate and no insolublehaemoglobin on filter paper. Centre: Patient with sickle cell trait(HbASj, showing pink-red filtrate and some red precipitate (HbS)on filter paper. Right: Patient wifh sickle cell anaemia showingpale yellow filtrate and heavy red precipitate on filter paper.

103 Haemoglobin C disease blood film. Note the many targetcells and occasional cell with HbC crystals. See 8.2.

Plates 98-103


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104 Blood film showing many ovalocytes and a fewelliptocytes

106 Red cells showing pale staining Hb H inclusionsas seen in a cresyl blue stained preparation. Cells havebeen likened to 'golf bails'. See 8.9.

108 Heinz bodies in red cells as seen in a crystalviolet stained preparation (can also be seen in areticulocyte stained preparation). See 8.9.


105 Beta thalassaemia major blood picture showing markedpoikilocytosis, hypochromasia, polychromasia, nucleated redcell, and target cells. The well haemoglobinized red cells are

transfused red cells. See 8.2.

107 Polychromasia with half-ghost cells and red cells withbitten out margins as seen in a G6PD deficient patient withhaemolytic anaemia due to oxidant stress. See 8.2.

109 Blood film showing small densely staining spherocytesand large polychromatic cell (reticulocyte). See 8. 7.

Plates 104-109


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district laboratory practice in tropical countries, color plates

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