A STUDY ON BACTERIOLOGY OF THE EYE INFECTIONS IN MAN …

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A STUDY ON BACTERIOLOGY OF THE EYE INFECTIONS IN MAN AND ANIMALS IN KHARTOUM STATE

By

IMAN BIREIR MOHAMMED TOM (B.V.SC., 1996)

University of Khartoum

Supervisor Prof: SULEIMAN MOHAMMED EL SANOUSI

Thesis submitted to the University of Khartoum in partial fulfillment of the requirements for the degree of Master of

Microbiology

Department of Microbiology Faculty of Veterinary Medicine

University of Khartoum March, 2008

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DEDICATION

TO MY FAMILY.... FATHER.... MOTHER.... BROTHERS.... AND SESTERS....

I DEDICATE THIS WORK

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TABLE OF CONTENTS

SUBJECT

DEDICATION

LIST OF TABLES

LIST OF FIGURES

ACKNOWLEDGEMENTS

ABSTRACT

ARABIC ABSTRACT

INTRODUCTION

CHAPTER ONE: LITERATURE REVIEW

1.1

The eye

1.1.1

Anatomy of the eye

1.1.2

Defense mechanisms of the eye

1.1.3

The tears

1.2

Eye infections

1.2

Major eye infections

1.2.1

Conjunctivitis

1.2.1.1 Bacterial conjunctivitis

1.2.1.1.1 Acute bacterial conjunctivitis

1.2.1.1.2 Chronic bacterial conjunctivitis

1.2.1.2 Chlamydial conjunctivitis

1.2.2

Keratitis (Corneal infection)

1.2.3

Keratoconjunctivitis

1.2.4

Infectious Bovine Keratoconjunctivitis (IBKC)

1.2.5

Eye lids infections

1.2.5.1 Blepharitis

1.2.5.2 Stye

1.2.5.3 Chalazion

1.2.6

Infections of the lacremal apparatus

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II X11 XI X XV XVI 1 3 3 3

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1.2.6.1 Dacryocystis

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1.2.6.2 Dacryoadenitis

14

1.2.6.3 Canaliculitis

14

1.2.7

Orbital infections

14

1.2.8

Endophthalmitis

15

1.2.9

Uvitis

15

1.2.6.1 Uvitis in horses

16

1.2.10 Retinitis

16

1.2.11 Skleral buckle infections

16

1.3

The normal flora of the eye

17

1.3.1

Human

17

1.3.2

Cattle

17

1.3.3

Goat

18

1.3.4

Camel

18

1.3.4

Poultry

18

1.3.4

Sheep

18

1.4

Microorganisms Associated with eye infections

19

1.4.1

Chlamydia spp

19

1.4.1.1 Chlamydia psittaci

19

1.4.1.2 Chlamydia trachomatis

20

1.4.2

Mycoplasma spp

20

1.5

Bacteria associated with eye infections

21

1.5.1

Gram-positive bacteria

21

1.5.1.1 Staphylococcus spp

21

1.5.1.2 Streptocuccus spp

27

1.5.1.3 Micrococcus spp

29

1.5.1.4 Corynebacterium spp

30

1.5.1.5 Actinomyces spp

32

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1.5.1.6 Propionibacterium spp

32

1.5.1.6 Listeria spp

33

1.5.1.7 Aerococcus spp

34

1.5.1.8 Enterococcus spp

34

1.5.1.9 Bacillus spp

34

1.5.1.10 Mycobacterium spp

35

1.5.1.11 Other Gram-positive bacteria associated with eye 36

infections

1.5.2

Gram-negative bacteria

36

1.5.2.1 Neisseria spp

36

1.5.2.2 Branhamella spp

37

1.5.2.3 Bordetella spp

38

1.5.2.3.1. Bordetella parapertussis

38

1.5.2.4 Moraxella spp

38

1.5.2.5 Pseudomonas spp

39

1.5.2.6 Burkolderia spp

40

1.5.2.7 Haemophilus spp

41

1.5.2.8 The Enterobacteria

42

1.5.2.9 Pasteurella spp

43

1.5.2.10 Aeromonas spp

43

1.5.2.11 Other Gram-negative bacteria associated with eye infections 44

1.5.3

Some antibiotics used in the treatment of eye infections

44

CHAPTER TWO: MATERIALS AND METHODS

47

2.1

Asepsis and Sterilization

47

2.1.1

Hot-air oven

47

2.1.2

Direct flame

47

2.1.3

Autoclaving

47

2.1.4

Tyndallization

47

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2.1.5

Irradiation and disinfectants

47

2.2

Media for isolation, identification and sensitivity testing 47

2.2.1

Solid media

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2.2.1.1 Nutrient agar

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2.2.1.2 Blood agar

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2.2.1.3 Chocolate agar

48

2.2.1.4 MacConkey's agar

48

2.2.1.5 Urea agar

49

2.2.1.6 Aesculin agar

49

2.2.1.7 Simmon's citrate agar

49

2.2.1.8 Kligler's iron agar (KIA)

50

2.2.1.9 Amonium salt sugars (ASS)

50

2.2.1.10 Casein agar

51

2.2.1.11 Starch agar

51

2.2.1.12 Bile agar

51

2.2.1.13 Nutrient agar containing4%, 6%, 8%, 10% Sodium chloride 51

2.2.1.14 Arginine agar

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2.2.1.15 Gelatin medium

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2.2.1.16 Dorset egg medium

52

2.2.1.17 Diagnostic sensitivity test (DST) agar

53

2.2.2

Semi-solid media

53

2.2.2.1 Hugh and Leifson's (O-F) medium

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2.2.2.2 Motility medium

53

2.2.3

Liquid media

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2.2.3.1 Nutrient broth

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2.2.3.2 Salt broth

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2.2.3.3 Peptone water

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2.2.3.4 Peptone water sugars

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2.2.3.5 MR-VP test medium

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2.2.3.6 Nitrate broth

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2.3

Reagents

55

2.3.1

Hydrogen peroxide

55

2.3.2

Oxidase test reagent

56

2.3.3

Kovacs' reagent

56

2.3.4

Voges-Proskauer test reagents

56

2.3.5

Methyl Red solution

56

2.3.6

Nitrate test reagents

56

2.3.7

Test papers with reagents

57

2.3.7.1 Lead acetate papers

57

2.4.1

Andrade's indicator

57

2.4.2

Bromothymol blue

57

2.4.3

Bromocresol purpel

57

2.4.4

Phenol red

57

2.5

McFarland 0.5 turbidity standard

57

2.6.1

Blood

58

2.6.2

Plasma

58

2.6

Samples of the study

58

2.6.1

Collection and transportation of samples

58

2.7

Culture methods

59

2.7.1

Primary inoculation on solid media

59

2.7.2

Subculture and purification of the isolates

59

2.7.3

Incubation of cultures

59

2.7.4

Examination of cultures

59

2.8

Identification of isolated bacteria

59

2.8.1

Primary tests

61

2.8.1.1 Microscopic examination

61

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2.8.1.1.2 Preparation of smears

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2.8.1.1.3 Staining

61

2.8.1.1.3.1 Gram's-stain

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2.8.1.1.3.2 Spore stain

61

2.8.1.2 Catalase test

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2.8.1.3 Oxidase test

62

2.8.1.4 Motility

62

2.8.1.5 Oxidation-fermentation (O-F) test

62

2.8.2

Secondary identification of bacteria

63

2.8.2.1 Nitrate reduction test

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2.8.2.2 Voges-Proskauer(V-P) test

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2.8.2.3 Sugar fermentation test

63

2.8.2.4 Urease test

63

2.8.2.5 Citrate utilization

64

2.8.2.6 Kligler's Iron Agar test (KIA)

64

2.8.2.7 Indole production

64

2.8.2.8 Methyl red test

64

2.8.2.9 Gelatin hydrolysis

65

2.8.2.10 Casein hydrolysis

65

2.8.2.11 Aesculin hydrolysis test

65

2.8.2.12 Bile solubility

65

2.8.2.13 Novobiocin sensitivity test

65

2.8.2.14 The CAMP test

66

2.8.2.15 Coagulase test

66

2.8.2.15.1 Slide method

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2.8.2.15.2 Tube method

66

2.8.2.16 Starch hydrolysis

67

2.8.2.17 Pigment production

67

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