Isolation of Aerobic Bacteria from Pneumonic Lung of Camels

102 Pages·2012·4.08 MB·English

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Isolation of Aerobic Bacteria from Pneumonic Lung of
Camels

By
Asmhan Fadl Alla Mohammed Elhilali
B. Sc. (2003)
Faculty of Veterinary Medicine
University of Khartoum

Supervisor
D. Awadelkarim A. Ibrahim

A thesis submitted to the University of Khartoum in partial fulfillment of the
requirements for the Degree of M. Sc. in Microbiology

University of Khartoum
Faculty of Veterinary Medicine
Department of Microbiology

January, 2012

I
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Dedication
To my parents, my brothers and sisters
To all whom I love

II
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III
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(12.7%) Coagulate- negative Staphylococci ϭ (8%) Staphylococcus aureusϭ
(17.8%) Streptococcus spp.ϭ(7.4%) Micrococcus spp.ϭ
(8.1%)Corynebacterium spp.ϭ(14.1) Actinomyces spp. (15.5%) Bacillus spp.ϭ
(3%) Escherichia coliϭ (3%) Klebsiella pneumoniaϭ (3.7%) Aeromonas
hydrophilaϭ (3.7) Proteus mirabilissϭ (1.5) Alcaligenes faecalisϭ(3.7%)
Kingella kingae.
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IV
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Abstract
This study was carried out with the aim of identifying aerobic bacterial
species involved in lung lesions of one -humped adult camels
(Camelusdromedarius) slaughtered at Elsalam abattoir enterprise, Khartoum
State .The study was conducted from February to May 2011.
All camels were originated from various pastoral areas. A total of 100 camel
lungs were inspected at random during the study period, of which 60 (60%)
possessed gross pulmonary lesions. All pneumonic lungs with lesions were
processed for bacteriology and bacterial growth was only obtained from
52(81.5%) of the pneumonic lung samples.
A total of 135 bacterial species were isolated and identified to the species
level using cultural characteristics and biochemical tests. These included
coagulase- negative staphylococci (12.7%), Staphylococcus aureus(8%),
Streptococcus spp. (17.8%), Micrococcus spp. (7.4%) Corynebacteria
spp.(8.1%), Actinomyces spp. (14.1) Bacillus spp. (15.5%) Escherichia coli
(3%) Klebsiella pneumonia (3%), Aeromonas hydrophila(3.7%), Proteus
mirabilis (3.7%) Alcaligenes faecalis(1.5%) Kingella kingae(3.7%). Some of
these are potential pathogens that could induce respiratory diseases and other
isolates might induce Respiratory diseases under stressful conditions.

V
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Table of contents
Page

DEDICATION II

ACKNOWLEDGEMENTS III

ARABIC ABSTRACT IV

ABSTRACT V

TABLE OF CONTENTS VI

LIST OF TABLES XVI

LIST OF FIGURES XVII

Chapter one 1

Introduction 1

Chapter two 3

Literature Review 3

2. 1 Introduction 3

2.2. Anatomy of camels lung 3

2.3. Respiratory tract infections in camels 4

2.4. Common bacteria associated with pneumonia 5

2.4.1. Gram positive bacteria 5

2.4.1.1. Staphylococci
5
2.4.1.1.1. Staphylococcus aureus
5
2.4.1.2. Micrococci 6

2. 4.1.3. Streptococci 6

VI
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2.4.1.3.1. Group B streptococci 7

a. Classification 7

b. Lance field's grouping 7

c. haemolysis production 7

2.4.1.4. Corynebacterium spp 7

2.4.1.5. Actinomycetes 8

2.4.1.6. Bacillus spp 8

2.4.2. Gram negative bacteria 9

2.4.2.1. Enterobacteriaceae 9

2.4.2.1.1. Escherichia coli 9

2.4.2.1.2. Klebsiella spp 9

2.4.2.1.3. Proteus spp 10

2.4.2.1.3.1. Proteus mirabilis 10

2.4.2.2. Aeromonas hydrophilia 11

2.4.2.3. Alcaligenes SPP 11

2.4.2.4. Kingella spp 11

2.5. The important bacterial respiratory diseases 11

2.5.1. Pasteurellosis 11

2.5.2. Tuberculosis 12

2.5.3. Mycoplasmosis 13

2.6. Bacteria isolated from infected respiratory tract of camels 13
VII
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2.7. Bacteria isolated from respiratory tract of apparently healthy 17
camels
Chapter three 19

Materials and methods 19

3.1. Study area 19

3.2. Collection of samples 19

3.3. Isolation of organism 19

3.4. Sterilization 20

a. Flaming 20

b. Red heat 20

c. Hot air oven 20

d. Moist heat 20

3.5. Reagents and Indicators 20

3.5.1.1. Sodium Chloride 20

3.5.1.2 Alpha-naphthol solution 21

3.5.1.3. Potassium hydroxide 21

3.5.1.4. Hydrogen peroxide 21

3.5.1.5. Kovac's reagent 21

3.5.1.6. Methyl red Solution 21

3.5.1.7. Tetramethyl-p-phenylaminedihydrochloride 21

3.5.1.8. Nitrate test reagent 22

VIII
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3.5.2. Indicators 22

3.5.2.1 Andrade's indicator 22

3.5.2.2 Bromothymol blue 22

3.6. Preparation of Culture Media 22

3.6.1. Liquid media 23

3.6.1.1. Nutrient broth 23

3.6.1.2. Peptone water 23

3.6.1.3. Peptone water sugar 24

3.6.1.4. Glucose- Phosphate medium 24

3.6.1.5. Nitrate medium 25

3.6.1.6. Serum Water sugars 25

3.6.2. Semi solid media 25

3.6.2.1. Motility test medium- Cragie tube medium 25

3.6.2.2 Oxidation-fermentation medium 26

3.6.3. Solid media 27

3. 6. 3.1. Ammonium salt sugars 27

3.6.3.2. Bile Esculin Azide Agar 27

3.6.3.3. Collection of blood for enriched media 28

3.6.3.4. Blood agar 28

3.6.3.5. MacConkey's agar 29

3.6.3.6. Milk agar 29
IX
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3.6.3.7. Nutrient agar 30

3.6.3.8. Nutrient Gelatin 30

3.6.3.9. Simmons citrate agar 31

3.6.3.10. Starch 31

3.6.3.11. Urea agar Base 32

3.7. Culture media 33

3.7.1. Blood agar 33

3.7.2. MacConkey agar 33

3.8. Isolation 33

3.9. Purification 33

3.10. Preservation 33

3.11. Microscopic Examination 34

3.12. Biochemical Tests 34

3.12.1. Sugar fermentation test 34

3. 12.2. Oxidase test 34

3.12.3. Catalase test 34

3.12.4. Coagulase test 35

3.12.5. The oxidation- fermentation test 35

``3.12.6. Indole production test 35

3.12.7. Methyl red test 36

X