E. Coli

A STUDY ON THE USE OF ANTIBIOTICS IN 

VITRO FOR SOME PATHOGENIC ENTERIC 

BACTERIA FROM DIFFERENT SOURCES 

A Thesis 

By 

FATMA ADEL ATTIA MOHAMED 

(B. Sc, Zagazig University, 2005) 

Under Supervision Of 

Prof. Dr. Ahmed Mohamed Ammar 

Professor of Microbiology 

Faculty of Veterinary Medicine 

Zagazig University 

Prof. Dr. Mohamed El-Bakry Abd El-rahim 

Professor of Microbiology 

Faculty of Veterinary Medicine 

Zagazig University 

Prof. Dr. Emad Rizkalla Zaki 

Chief Researcher of Bacteriology 

Head of Buffalo Diseases research Department 

Animal Health Research Institute-Dokki-Giza 

A Thesis 

Submitted to Zagazig University, Faculty of Vetrinary Medicine 

Department Of Bacteriology, Mycology and Immunology For Master 

degree of Bacteriology 

(2008)

ﺾﻌﺒﻟ ﺎﻴﻠﻤﻌﻣ ﺔﻳﻮﻴﺤﻟا تادﺎﻀﻤﻟا ماﺪﺨﺘﺳا ﻰﻠﻋ ﺔﺳارد 

ةدﺪﻌﺘﻣ 

ردﺎﺼﻣ ﻦﻣ ﺔﻳﻮﻌﻤﻟا ﺎﻳﺮﺘﻜﺒﻟا 

ﺪﻤﺤﻣ 

ﻦﻣ ﺔﻣﺪﻘﻣ ﺔﻟﺎﺳر 

ﺔﻴﻄﻋ لدﺎﻋ ﺔﻤﻃﺎﻓ 

2005 ﻖﻳزﺎﻗﺰﻟا ﺔﻌﻣﺎﺟ , مﻮﻠﻌﻟا ﺔﻴﻠآ , مﻮﻠﻌﻟا سﻮﻳرﻮﻟﺎﻜﺑ 

فاﺮﺷا ﺖﺤﺗ 

رﺎﻤﻋ ﺪﻤﺤﻣ ﺪﻤﺣا/. 

د. 

ا 

ﺎﻴﺟﻮﻟﻮﻴﺑوﺮﻜﻴﻤﻟا ذﺎﺘﺳا 

ﻖﻳزﺎﻗﺰﻟا ﺔﻌﻣﺎﺟ -ىﺮﻄﻴﺒﻟا 

ﺐﻄﻟا ﺔﻴﻠآ 

ﻢﻴﺣﺮﻟا ﺪﺒﻋ ىﺮﻜﺒﻟا ﺪﻤﺤﻣ /. د. 

ا 

ﺎﻴﺟﻮﻟﻮﻴﺑوﺮﻜﻴﻤﻟا ذﺎﺘﺳا 

ﻖﻳزﺎﻗﺰﻟا ﺔﻌﻣﺎﺟ -ىﺮﻄﻴﺒﻟا 

ﺐﻄﻟا ﺔﻴﻠآ 

ﻰآز ﺔﻠﻟا قزر دﺎﻤﻋ/. 

د. 

ا 

ﻰﺟﻮﻟﻮﻳﺮﺘﻜﺒﻟا ثﻮﺤﺑ ﺲﻴﺋر 

سﻮﻣﺎﺠﻟا ضاﺮﻣا ثﺎﺤﺑا ﻢﺴﻗ ﺲﻴﺋرو 

ةﺮهﺎﻘﻟا- 

ﻰﻗﺪﻟا-ناﻮﻴﺤﻟا 

ﺔﺤﺻ ثﻮﺤﺑ ﺪﻬﻌﻣ 

ﻰﻟا 

ﺔﻋﺎﻨﻤﻟاو تﺎﻳﺮﻄﻔﻟاو ﻰﺟﻮﻟﻮﻳﺮﺘﻜﺒﻟا ﻢﺴﻗ , ىﺮﻄﻴﺒﻟا ﺐﻄﻟا ﺔﻴﻠآ , ﻖﻳزﺎﻗﺰﻟا ﺔﻌﻣﺎﺟ 

ﻰﺟﻮﻟﻮﻳﺮﺘﻜﺒﻟا ﻰﻓ ﺮﻴﺘﺴﺟﺎﻤﻟا ﺔﺟرد ﻰﻠﻏ لﻮﺼﺤﻠﻟ 

( 

2008)

List of contents 

Page 

1-Introduction………………………………..…………….1 

2-Review of literature 

2.1. Incidence of pathogenic Enterbacteriaceae…………….…3 

2.1.1. Different E. coli serotypes isolated from broilers…….3 

2.1.2. Salmonella…………………………………………….9 

2.1.3. Frequent of Enterobacteriaceae in broiler and 

surrounding environment………………………..…………15 

2.3. Pathogenic Enterbacteria in Human………………..….. 19 

2.3.1. E. coli…………………………………………..……22 

2.3.2. Salmonella……………………………………..….....27 

3-Materials and Methods 

3.1. Materials……………………………………….……....29 

3.1.1. Samples: 

3.1.2. Media used: 

3.1.3. Media for biochemical reactions………….. ….….….31 

3.1.4. Solutions and indicators in biochemical identification.34 

3.1.6. Media used for antibiotic sensitivity test………….…..35 

3.1.7.Diagnostic E.coli Antisera……………………….…....35 

3.2. Methods 

3.2.1. Collection of sample…………………………….……38 

3.2.2. Isolation of Enterbacteriaceae from organ samples…...38 

3.2.3. Bacteriological identification………………………....39 

3.2.4. Biochemical characterization………………………....39 

3.2.5. Serological typing of the isolated E. coli…………..…41 

3.2.6. Antibiotic sensitivity test………………………………42

4-Results 

4.1-Enterobateria in chicken specimens……….…………....45 

4.2- Enterobacteria in Human urine ………………………….50 

4.3-Biochemical identification of chicken Enterobacteria…....50 

4.4- Biochemical identification of urine isolates………….…52 

4.5-Pathogenicity test………………………..………….…….55 

4.6-Results of serotyping of E.coli isolates. ..……….……….55 

4.6-Results of antibiogram on biotypes from one day 

old chicks…………………………………………………….55 

4.7-Sensitivity pattern of the isolates from one day old chick..63 

4.8-Results of antibiogram on layer isolates………………....65 

4.9-Sensitivity Pattern of layer isolates…………..………….65 

4.10-Results of antibiogram on isolates from broiler……..…68 

4.11Sensitivity pattern of broiler isolates………..…………...70 

4.13-Sensitivity percentages of each antimicrobial agent.……72 

4.12-Results of antibiogram on biotypes from human urine...76 

5-Discussion ………………………………………...……..87 

6-Summary ……………………………………….…..…...91 

7-Conclusion…………………………………….……..….96 

8-References………………………….……….……..…….98 

Arabic summary 

List of Tables

Tables 

Page 

Table(1)Biochemical reactions, members of Enterobactericeae.36 

Table(2)Standard zone of inhibition of the used antibiotics 

for antimicrobial sensitivity test……………………..………..37 

Table(3)Prevalence rate of isolated Enterobacteria from different 

Sources……………………………………………………..….….45 

Table(4)Incidence of chicken Enterobacteria obtained on 

MacConkeyۥs medium from broiler…..…………………….……47 

Table (5) Incidence of chicken Enterobacteria obtained on 

MacConkeyۥs medium from one day old chicks and layers…..….48 

Table(6)Incidence of isolates after identification according to 

biochemical reactions…….………………………………...…….51 

Table(7)Antibogram of isolates from one day old chicks against 10 

chemotherapeutic agents………………….……………..…….…60 

Table (8) Sensitivity pattern of the isolates from one day old 

chicks against 10 chemotherapeutic agents……………….….….63 

Table (9) Antibiogram of layer isolates against 11 chemotherapeuticagents…………………………………………….…..65 

Table(10)Sensitivity pattern for isolates from layers……………68 

Table(11)Antibiogram of isolates from broiler against 11 chemo- 

therapeutic agents………………………………………….……..69 

Table(12)Sensitivity Pattern of isolates from broiler…………....72 

Table (13) Sensitivity percentages of each antimicrobial against 

isolates from broiler, One day old chicks and layers……….……74 

Table (14) Antibogram E.coli isolates from human urine………76

List of Figures 

Figures Page 

Fig.(1)Tube lactose fermentation using phenol red indicator 

(yellow in positive test but faint red for control medium)…..…….49 

Fig.(2)Xylose lysine decarboxylase (XLD)medium, only in the 

fourth tube (right side) the isolate fermented sugars producing 

yellow colour, no H2s was produced……….……………………..53 

Fig.(3)Triple sugar iron agar(TSI), third and fourth tubes(right 

side),Acid slant(yellow) over Acid butt(yellow) no H2S. with 

gas in the third tube. First and second are control…… ………..…54 

Fig.(4)Citrate negative E.coli isolate……………………………...56 

Fig.(5)Pathogenicity lesions showing congested liver and peri- 

charditis after challenge with 5.8X105 fresh E.coli culture..……..57 

Fig.(5)Pathogenicity lesions showing congested liver and 

Pericharditis………….……..…………………………….……..…57 

Fig.(6)An isolate was sensitive to gentamycin, ciprofloxacin 

and nitrofurantoin..……………………….………………….…….58 

Fig.(7)An isolate was sensitive to ciprofloxacin, amikacin 

and gentamycin…………….……………………………..……….59 

Fig.(8)Sensitivity percentages of antimicrobials against broiler, 

one day old chicks and layer Isolates…………..…………………75

Acknowledgements 

First of all thanks to ALLAH to whome I relate any success in my 

life. 

I would like to thank Prof. Dr. Ahmed Mohamed Ammar 

Professor of Microbiology, Faculty of Veterinary Medicine, Zagazig 

University for his help, advice encouragement during the entire 

work. 

I would like to thank Prof. Dr. Mohamed El-Bakry Abd-Elrahim 

Professor of Microbiology Faculty of Veterinary Medicine,Zagazig 

University for his advice and great support throughout this work. 

I would like to deep thank to Prof. Dr.Emad Rizkalla Zaki, Chief 

Researcher of Bacteriology, Head of Buffalo Diseases research 

Department Animal Health Research Institute-Dokki-Giza for his 

technical support. 

Finally I would like to thank all staff members of Bacteriology, 

Mycology and Immunology department for their help and advice.

1. INTRODUCTION 

Introduction 

Group of Enterobacteriaceae in human includes several that cause 

primary infections of the human gastrointestinal tract. Bacteria that 

affect the gastrointestinal tract include certain strains of E. coli and 

Salmonella, Shigella, and Yersinia entercolitica. Members of this 

family are major causes of opportunistic infection (including 

septicemia, pneumonia, meningitis and urinary tract infections). 

Examples of genera that cause opportunistic infections are: 

Citrobacter, Enterobactcr, Escherichia, Hafnia, Morganella, 

Providencia and Serratia. Selection of antibiotic therapy is complex 

due to the diversity of organisms. The mortality and morbidity are 

much higher for the ages less than two years than in older one, fever, 

sever loss of electrolyte and dehydration accompanied with watery to 

bloody diarrhea (Forfor and Arneil, 1978). 

Many species are intestinal pathogens or commensals in the 

intestine of man and animal, few are saprophytes in soil and water. 

Some species are also transmitted between man and animals (WHO, 

2000). 

The most common serotypes are E. coli O157:H7 and E. coli 

O26:H11. These are unique from the enteropathogenic E. coli (EPEC) 

organisms that are the main causes of infantile diarrhea (Shebib et al., 

2003). 

Avian pathogenic E.coli (APEC) cause aerosacculitis, poly- 

serositis, septicemia in chicken. APEC isolates commonly belong to 

1

2 

Introduction 

certain serogroups, O1, O2, and O78 (Dho-Moulin and 

Fairbrother,1999). 

Poultry is one of the most reservoir of salmonellosis causing great 

losses and hazards to public health (El-Sayed, 1997). 

Microbial species and strains have different degrees of 

susceptibility to different chemotherapeutic agents. Moreover, the 

susceptibility of a microorganism can be changed with time even 

during therapy with a specific drug. Thus a specialist must know the 

sensitivities of the pathogen before treatment to be started (Tortora et 

al.,2002). 

Aim of the work 

The purpose of this work is to achieve the following points: 

1-Isolation and biochemical identification of pathogenic bacteria 

of Enterobacteriaceae group. 

2- Serotyping of the isolated bacteria . 

3-Study the sensitivity of isolates to antimicrobial substances 

using disc diffusion method. 

4- Study the pathogenicity of isolates in broiler.

3.1. Materials: 

3.1.1. Sample: 

3.MATERIAL AND METHODS 

A total of 295 samples obtained from livers, spleen, heart of 

freshly died broiler, one day old chicks and layers in Dakahlia 

province. 

A total of 53 children, male and female urine samples were 

obtained from human private laboratories. 

3.1.2. Media used: 

3.1.2.1. Selective enrichment broth: 

Selenite F broth (Edward and Ewing, 1972) 

Bacteriological peptone 5.0 g/l 

Lactose 4.0 g/l 

Sodium phosphate 10.0 g/l 

3.1.2.2. Selective culture media: 

3.1.2.2.1. Nutrient Agar (Oxoid) 

Lab-lemco powder 1.0 g/l 

Yeast extract 2.0 g/l 

Peptone 2.0 g/l 

Sodium chloride 5.0 g/l 

Agar 

pH 7.4 

15.0 g/l 

3.1.2.2.2. Blood Agar (Oxoid) 

Lab-lemco, powder 10.0 g/l 


29


Agar 

pH 7.3 

15.0 g/l 

3.1.2.2.3. MacConkey's Agar (Oxoid): 



Bile salts 5.0 g/l 


Neutral red 0.075 g/l 

Agar 

pH 7.4 

12.0 g/l 

3.1.2.2.4. Salmonella Shigella Agar (Oxoid): 




Bile salts 5.5 g/l 

Sodium citrate 10.0 g/l 

Sodium thiosulphate 8.5 g/l 

Ferric citrate 1.0 g/l 

Brilliant green 0.00033 g/l 

Neutral red 0.025 g/l 

Agar 

pH 7.3 

12.0 g/l 

3.1.2.2.5. Levins Eosin Methylene Blue (Oxoid) 


30


Dipotassium hydrogen phosphate 2.0 g/l 

Eosin Y 4.0 g/l 

Methylene blue 0.065 g/l 

Agar 

pH 6.8 

15.0 g/l 

3.1.3.Media used for Biochemical examination: 

3.1.3.1. Peptone water media (oxoid) 


Sodium chloride 

pH 7.2 

5.0 g/l 

It used for detection of indole production but now better results 

can be obtained by the used of tryptone water (oxoid). 

3.1.3.2. Glucose-phosphate peptone water medium (Oxoid) 


Dextrose 5.0 g/l 

Phosphate buffer 

pH 7.5 

5.0 g/l 

This medium is used for methyl-red and Voges-Proskauer tests 

for differentiation of the coli-aerogenes group. 

3.1.3.3. Simmons citrate (Oxoid) 

Magnesium sulphate 0.2 g/l 

Ammonium dihydrogen phosphate 0.2 g/l 

Sodium ammonium phosphate 0.8 g/l 

31

Sodium citrate, tribasic 2.0 g/l 


Bromothimol blue 0.08 g/l 

Agar 

pH 7.0 

15.0 g/l 

It used for citrate utilization ability of the isolated 

microorganisms. 

3.1.3.4. Urea agar medium (Oxoid): 




Disodium phosphate 1.2 g/l 

Potassium dihydrogen phosphate 0.8 g/l 

Phenol red 0.012 g/l 

Agar 

pH 6.58 

15.0 g/l 

It is recommended for preparation of Christensen medium for 

detection of urea splitting organisms such as proteus vulgaris. 

3.1.3.5. Nutrient gelatin (Oxoid): 



Gelatin 

pH 6.8 

120.0 g/l 

This media used for determination of gelatin liquification, which 

is characteristic for proteolytic bacteria. 

32

3.1.3.6. Triple Sugar Iron Agar Medium TSI (Oxoid) 


Yeast extract 3.0 g/l 




Sucrose 10.0 g/l 


Ferric citrate 0.3 g/l 

Sodium thiosulphate 0.3 g/l 

Phenol red 9.5 g/l 

Agar 

pH 7.4 

12.0 g/l 

3.1.3.7. Sugar media 

Glucose, lactose, sucrose, mannitol, maltose, salicin, dulcitol, 

galactose and fructose were included in peptone water. 

Peptone 1% 

Sodium chloride 0.5% 

Phenol red Indicator 5% (0.2 gm%) 

Sugar 1% 

Durham tubes were used for studying the fermentative ability of 

the isolated microorganisms. 

33

3.1.4.Solutions and indicators in biochemical 

identification (Cruickshank et al., 1975): 

3.1.4.1. Oxidase reagent (Bio-Merieux, 1980) 

Freshly prepared 1%solution of tetra methyl-p-phenylene diamine 

dihydrochloride was used to test for oxidase production. 

3.1.4.2. Kovac's reagent: 

Pdimethylaminobenzaldhyde 10 g 

Amyl alcohol 150 ml 

Concentrated HCl 50 ml 

3.1.4.3. Methyl Red Reagent: (M.R) 

Methyl red 0.1 g 

Ethyl alcohol 95% 300.0 ml 

Distilled water 200.0 ml 

3.1.4.4.Voges-Proskauer (V.P.) test reagents: 

To 5ml of a 48-hours at 37°C, 3ml of alcoholic solution of naphthol 

and 1ml of 40% of potassium hydroxide solutions were 

added. Pink colored ring developed withen 10-15 min. while negative 

if there is no pink ring. 

3.1.4.5. Urea solution (40%) (Oxoid, sr20) 

3.1.4.6. H2S production: 

Triple sugar iron (T.S.I) (oxoid) examination was done daily up 

to the 7 th day for detection of H2S production as well as for gas 

production. 

34

3.1.4.7. Nitrate reduction test reagents: 

Reagent A: -Naphthyl amine 5 g..m 

5N acetic acid 30% 1 litter 

Reagent B: Sulfanilic acid 8 g. 

3.1.5. Stains used: 

(1975). 

5N acetic acid 30% 1 litter 

Modified Gram’s stain used as described by Cruickshank et al., 

3.1.6. Media used for antibiotic sensitivity test 

3.1.6.1. Media used: 

Mueller-Hinton agar (oxoid) was prepared according to Finegold 

and Martin (1982). 

3.1.6.2. Antimicrobial sensitivity discs (Din, 1998): 

Antmicrobial susceptibility discs were obtained from BioMerieux 

as follows in table (2). 

3.1.7.Diagnostic E.coli Antisera 

The isolates were identified serologically by diagnostic E.coli 

antisera for pathogenic types were used in this study. The diagnostic O 

sera 51 vials (polyvalent,8 vials and monovalent,43 vials (product code 

312001) 

Polyvalent 1: O1 , O26 , O86A , O111 , O119 , O127A , O128 

Polyvalent 2: O44 , O55 , O125 , O 126, O146 , O166 

Polyvalent 3: O18 , O114 , O142 , O151 , O157 , O185 

35

Polyvalent 4: O6 , O27 , O78 , O148 , O159 , O168 

Polyvalent 5: O20 , O25 , O63 , O153 , O 167 

Polyvalent 6: O8 , O15 , O115 , O169 

Polyvalent 7: O28ac , O112AC , O124 , O136 , O144 

Polyvalent 8: O29 , O143 , O152 , O164 

36

3.2. Methods 

3.2.1. Collection of sample: 

The birds were collected every week, starting from 8 days of a 

arrival till the marketing age 38 day old. The source of birds was from 

18 broiler farms in Dakahlia governorate. 

3.2.2. Isolation of Enterbacteriaceae from organ samples: 

Firstly, chickens were scarified, dipped in phenol 5%solution, 

opened aseptically for bacteriological examination, the macroscopic 

lesions were sterilized using preheated scalpel. Then separate loopfuls 

were taken from heart blood, liver, lung, gall bladder (Siam, 1998). 

Each separate loopful was directly inoculated into separate nutrient 

broth tubes, then subcultured into MacConkey broth and MacConkey 

agar. The inoculated broth and the streaked agar media were incubated 

at 37°C for 24-48 hours pure colonies were picked up and preserved 

onto slope agar for further biochemical and serological identification 

according to Edwards and Ewing (1972), Finegold and Martin 

(1982), Krieg and Holt (1984) and Holt et al., (1999). 

3.2.3. Isolation of Enterobacteriaceae from urine samples 

Urine samples were collected from private laboratories. Firstly, 

isolation was done by soaking a cotton swap in the specimen then 

streaked on nutrient agar media then it was incubated in incubator at 

37°C for 24 hours. After growth the pure colonies were picked up and 

streaked on MacConkey agar media and incubated at 37°C for 24 

hours then the suspected colonies were picked up and streaked onto 

nutrient agar plates for purification then subjected to biochemical and 

37

serological identification according to Edward and Ewing (1972), 

Fine gold and Martin (1982), Krieg and Holt (1984) and Holt et 

al., (1999). 

3.2.3. Bacteriological identification 

3.2.3.1. Morphological characteristics: 

From suspected purified colonies, bacterial smears were prepared 

and stained with Gram’s stain and examined microscopically for 

morphological characteristics. 

3.2.3.2. Cultural characteristics: 

The colonial appearance was studied to investigate their 

structure, surface, edge, color, opacity, haemolysis of blood agar and 

pigment production. The motility of each isolate was detected by its 

stabbing into semisolid agar. The motile one grows away from line of 

stabbing. 

3.2.4. Biochemical characterization: 

The following methods of biochemical tests used for 

identification were carried out according to the schemes described by 

Cruickshank et al., (1975), Edwards and Ewing (1972), Finegold 

and Martin (1982), Kerig and Holt (1984) and Holt et al., (1999). 

1- Oxidase test 

A piece of filter paper was soaked with few drops of oxidase reagent. 

A colony of the test organism was then smeared onto the filter paper. If the 

organism is oxidase producing, the phenylene diamine in the reagent will 

38

e oxidized to a deep purple colour within few seconds (10 seconds), 

ignore any blue- purple colour that develops after 10 seconds. 

2- Indole test 

To 48 hours culture in 1% peptone water after incubation at 37°C, 0.5 

ml of Kovacۥs reagent was gently trickled down the side of the tube, 

development of a rosy colour ring indicates the presence of indol. 

3-Methyl red test 

Five drops of methyl red reagent were added to 5ml of 48 hours 

incubated glucose phosphate broth. Appearance of red colour indicates 

positive reaction but yellow colour indicate negative one. 

4- Voges- Proskauer test (V.P.) 

Three ml alcoholic solution of 40% alpha-naphthol were added to 1 

ml of glucose phosphate broth culture incubated at 37°C/48 hours. the 

mixture was thoroughly shaken and examined after 15 minutes to one hour. 

The presence of strong red colour indicates positive reaction. 

4- Citrate utilization test 

The ability to utilize citrate was detected by making a single streak 

over the surface of simmonۥs citrate agar slope and incubate it at 37°C for 7 

days. Development of blue colour indicates citrate utilization. 

5- Gelatin liquefication 

Gelatin medium tubes were inoculated with isolates and incubated at 

37°C for up 14 days. Tubes were examined every 2 days for liquifaction. 

39

6- Urease test 

The test determine the ability of bacteria to decompose urea by means 

of urease enzyme. The development of red colour indicates hydrolysis of 

urea. 

7- Sugar fermentation test 

The isolated organisms were cultured into sugar media containing 1% 

of the tested sugar. Phenol red act as indicator. Yellow colour indicate acid 

production while gas production seen in Durham’s tubes, after incubation 

at 37°C for 1-7 days. 

3.2.4. Serological identification of E.coli : 

Serological analysis was performed according to methods described by 

Edwards and Ewing(1972). 

1- O antigen group screening: 

a-Slide agglutination test: 

A glass pencil was used to divide a glass slide into 2 parts, one drop of 

the polyvalent sera and one drop of the polyvalent sera and one drop of the 

physiological saline were separately placed in the 2 section. 

Each isolate was densely suspended in physiological saline. One 

drop of the suspension, using the platinum loop, was placed in each 

separate section of the slides in the vicinity of polyvalent sera and 

physiological saline. The antigen and the serum drop were mixed in each 

section. 

The glass slides were titled back and front only strong agglutinations 

occurring within one minute were accepted as positive, when positive 

reaction was observed with one of the polyvalent sera, slide agglutination 

40

was carried out as it was described above by using monovalent sera 

comparing the polyvalent serum which agglutinated. 

B- Slide agglutination test using heated cells 

When one monovalent serum showed agglutination, the live cell 

suspension densely suspended in physiological saline should be heated at 

100°C for 1 hour, or at 121°C for 15 minutes to check that the serum 

produces agglutination of the heated cells as well. The absence of 

spontaneous agglutination was also checked using physiological saline. 

When both the live and the heated cells of a bacterium whose 

biochemical characteristics had been observed as identical to those of 

E.coli agglutinated in the reaction with one of the monovalent sera, the 

organism was regarded as pathogenic E.coli belonging to the serogroup 

representing the monovalent serum. However, only live cells were 

agglutinated(and heated did not) such a case was excluded from the 

interpretation. 

3.2.5. Antibiotic sensitivity test (Blair et al., 1970) 

A disc diffusion technique adapted as it was recommended by using 

pure culture from the tested isolates as follows: - 

1. Mueller Hinton agar plates were dried in the indicator until the surface 

will free from visible moisture. 

2. Few colonies of the tested organism were suspended in broth and 

incubated for 4-5 hours until the turbidity was seen (this turbidity will 

adjusted to much McFarland No .0.5 Barium sulphate standard tube 

(0.5ml of 1.175% barium chloride hydrate, to 99 ml of 1% sulphoric 

acid) By sterile Pasteur pipette, 1 ml of the suspension was inoculated 

into the surface of the plate. 

41

3. The plate was swabbed in different directions to wet the whole of its 

surface 

4. Excessive fluid was discarded by its pipetting then plates were dried 

for up to 30 min. 

5. The chosen antibiotic discs were distributed to the surface of the plate 

with sterile point forceps, a gentle pressure over the discs was done to 

ensure full sticking of the discs to the medium then the plates were 

aerobically incubated at 37° C /24 hours. 

6. The degree of the sensitivity was determined by measuring the 

diameter of inhibition zone in mm produced by diffusion of antibiotics, 

from disc to surrounding medium. 

Table(1)Biochemical reactions,members of Enterobactericeae, 

Cruickshank et al., (1975). 

M.O. 

E.coli 

Indol 

M.R 

V.P. 

Citrate 

Urease 

Lactose 

Maltose 

Mannitol 

Sucrose 

TSI 

+ + - - - + + + d Y/Y/H2S- 

Salmonella - + - + - - + + - R/Y/H2S 

+ 

Enterobacter 

aerogens 

- - + + - + + + + Y/Y/H2S- 

Citrobacter 

diversus 

+ + - + + d + + - Y/Y/H2S- 

Klebsiella 

pneumoniae 

- - + + + + + + + Y/Y/H2S- 

Edwadsiella 

tarda 

+ + - - - - + + + Y/Y/H2S- 

TSI=triple iron agar d=26-75%(positive), Y=yellow ,R=red 

42

Table(2):Standard zone of inhibition of the used antibiotics for 

antimicrobial sensitivity test 

Chemotherapeutic Agents Concentration 

43 

*Zone of inhibition 

R I S 

ciprofloxacin 10 mg 15 16-20 21 

cefotaxime 14 mg 16 15-22 23 

Amikacin 30 mg 14 15-16 17 

ampicillin 10 mg 13 14-16 17 

Gentamycin 10 mg 12 13-14 15 

Trimethoprime-sulpharnethoxazole 1.5+23.75 mg 10 11-15 16 

Colistin 10 mg 16 - 18 

Amoxycillin clavulanic 20/10 mg 13 14-17 18 

doxycyclin 30UI 12 13-15 16 

erythromycin 15 13 14-22 23 

rifampicin 5 16 17-19 20 

streptomycin 10 11 12-14 15 

R: Resistant. I: Intermediate. S: Sensitive. 

*: Zone of inhibition was measured by mm diameter.

2.REVIEW OF LITERATURE 

2.1. Incidence of pathogenic Enterbacteriaceae. 

Review of Literature 

2.1.1. Different E. coli serotypes isolated from broilers: 

Bozorgmehri et al. (1980) isolated 90 strains of E.coli from 

chickens and recorded that the registrated serotypes were O78: K80, 

O111:B4, O128:B12, O119:B14 and O86:B7 while the remaining 

strains could not be serologically typed. 

Burkhanova (1980) isolated 337 strains of enteropathogenic E. 

coli from blood, bone marrow and liver of dead baby chicks. He 

proved that the prevalent serogroups of E. coli were O:9, O:119, O:55, 

O:125, O:26 and O:78. 

Abd El-Galil et al. (1983) isolated E.coli from first 10 days old 

dead baby chicks of a private poultry farms in Sharkia Government. 

They were 30 cases with an incidence of 15% and the most prevalent 

pathogenic serogroups were O:125, O:119, O:78 and O:55. 

Joya et al. (1990) investigated two outbreaks of diarrhea in 

broiler chicks at two independent farms in the Philippines from, which 

no pathogens other than E.coli were found. 

Osman (1992) examined 150 bacteriological swabs, which were 

taken directly from liver, spleen, lung and heart blood of first ten days 

old chicks, which were collected from various localities in Sharkia 

Governorate broiler farms. Forty two E.coli strains were isolated with 

3


an incidence of 28% serological typing of the isolated E.coli strain 

refer to 13 of O125:B76, 8 of O119:B69, 5 of O78:B80, 4 of O55: 

B59 and 12 untypable. 

Abu-Elyazeed et al. (1995) stated that enterotoxigenic 

Escherichia coli (ETEC) are diverse pathogens that express heat-labile 

(LT) and/or heat-stable (ST) enterotoxins. 

Draz (1996) collected 60 water and 45 litter samples from 

poultry environment and poultry farms for bacteriological 

examination. E.coli was isolated with the frequency of 36.7% (22/60) 

and 17.8% (8/45) respectively. E.coli stains isolated from water and 

litter were serotyped as O:2, O:4, O:6, O:11, O:15 and O:26 with of a 

frequency in water, 1 (4.6%), 3 (13.6%), 4 (18.1%), 1 (4.6%), 1 

(4.6%) and 2 (9.0%) respectively. Meanwhile, they were in litter, 2 

(25%), 1 (12.5%), 1 (12.5%), 0 (0.0%), 1 (12.5%) and 0 (0.0%), 

respectively. 

Jordan and Pattison (1996) stated that certain E.coli serotypes 

could cause disease in poultry as yolk sac infection, coligranuloma 

(Hijarres disease), egg peritonitis and colisepticemia. Most of 

pathogenic E.coli belong to small range of serotypes, which include 

O78:K80, O1:K1 and O2:K1. Colibacillosis characterized by air 

sacculitis, peritonitis, perihepatitis, pericarditis and congested dark 

liver as well as congested lung. 

Barnes and Gross (1997) reported that the presence of E.coli in 

drinking water is considered indicative of faecal contamination. It was 

4


suggested that E.coli spread rapidly after hatching. Feed is often 

contaminated with pathogenic coliform but these can be destroyed by 

hot pelliting process. Rodent dropping often contain pathogenic 

coliforms. 

Blanco et al. (1997) established a study for detecting the 

serogroups of E.coli that causes avian colibacillosis in spain. The 

serogroups of 625 avian E.coli isolated between 1992-1993 were 

determined. The 458 E.coli from chickens with septicemia belonged 

to 62 different 0 serogroups; however, 59% were of 18 serogroups (O: 

1, O:2, O:5, O:8, O:12, O:14, O:15, O:18, O:20, O:53, O:78, O:81, 

O:83, O: 102, O:103, O:115, O:116 and O:132). The high prevalence 

of O:18, O:81, O:115, O:116, O:132 isolates was not expected and 

many indicate the emergence of five new serogroups associated with 

avian colibacillosis not yet reported. 

El-Morsi (1998) examined twenty five liver samples from 

poultry and found that 5 samples were positive to E.coli by incidence 

of 20%. The isolated serotypes of E.coli from liver samples were 2 

untypable (40%), 2 belonged to O111:K58 (40%) and one was O126: 

K71 (20%). 

Fisher et al. (1998) induced E. coli septicemia in broilers in 

order to determine if lesions of acute septicemia could be grossly 

detected in visceral organs of broiler carcasses. Increased spleen and 

liver weight were observed during the acute phase of septicemia. Air 

saculitis, pericarditis and periphepatitis were observed also during the 

acute phase. 

5


Sakr (1998) found that out of 572 intestinal samples from broiler 

chickens, 115 isolates were recovered 45 E.coli (7.7%). 

Siam (1998) isolated E.coli from broiler organs (920 birds) 

including heart blood, spleen, lung, liver and gall bladder during spring, 

autumn, winter and summer seasons with the percentage of 38.8, 35.5, 

26.008 and 20.55 respectively. The percentage of isolation of E.coli 

from heart blood was 27.17% followed by lung 15.2%, then gall 

bladder (12.39%), liver (10.8%) and lastly yolk sac (2.6%). E.coli was 

isolated from broiler organs in high incidence at 5 th week of age. 

Dho-Moulin and Fair brother (1999) stated that avian 

pathogenic E.coli (APEC) cause aero-sacculitis disease in chickens. 

APEC are found in the intestinal microflora of healthy birds and most 

of the diseases associated with them are secondary to environmental 

host predisposing factors. APEC isolates commonly belong to certain 

serogroup, O:1, O:2 and O:78. Experimental infection studies have 

shown that the air-exchange regions of the lung and the air sacs are 

important sites of entry of E.coli into the blood stream of birds during 

the initial stages of infection. 

El-Shamy (1999) carried out bacterial studies on agents causing 

enteritis in chicken breeder flocks. Fourty four diseased hens were 

collected from two large farms in Dakahlia province having diarrhoea 

and the samples were from intestine, heart blood, liver and spleen. 

One hundred and eleven E.coli were isolated with an incidence of 

18%. Serological identification of 40 strains revealing 15 strains O55: 

6


K5 (37.5%), 15 strains O111: K4 (37.5%), 6 strains O128: K6 

(15.0%) and 4 strains O88: K4 with a percentage of 10.0%. 

Hirsh and Zee (1999) illustrated that colibacillosis of fowl is an 

economically important disease caused by invasive strains of E.coli. 

the disease takes many forms in fowl depending upon the age of the 

host and mode of infection. The egg surface could be contaminated 

with potentially pathogenic strains at the time of laying. The bacteria 

penetrate the shell and infect the yolk sac. Embryo that survive may 

die shortly after hatching, with losses occurring as late as 2 weeks 

after hatching fowl may also be infected by the respiratory tract and 

develop respiratory or septicemic disease. The course may be rapidly 

fetal or chronic, manifested by debilitation, diarrhea and respiratory 

distress. Other clinical syndromes seemingly caused by E. coli include 

cellulites, synovitis, pericarditis, salpingitis and panophthalmitis. 

Gomis et al. (2000) examined 241 broiler birds in Srilanka for 

bacteriological examination from different gross lesions, 162 E.coli 

were recovered. Twenty one percent of the birds had multiple lesions 

due to E.coli. The frequency of detection of these lesions were, 162 

(67%) pericarditis, 26 (11%) air saculitis, 24 (10%) hepatitis, 12 (5%) 

perihepatitis and 16 (7%) polyserositis. Serogroups O:78, O:85 and O: 

88 were distributed among the 32% of typable E.coli. 

El-Sayed et al. (2001) isolated pathogenic E.coli from 50 

samples of poultry carcasses at shops in Mansoura city, Dakahlia 

province positive E.coli samples were 15 with an incidence rate 

30.5%. E. coli isolates were serotyped as, 6 strains O55: K59, 3 strains 

7


O78:K80, 2 strains O126:K71 and one strain was O114:K96, 3 

untypable strains with percentage of 12, 6, 6, 4 and 2% respectively. 

Gomis et al. (2001) characterized virulence factor of E.coli 

isolates from broiler with cellulites and other colibacilosis lesions E. 

coli derived from cellulities lesions produced virulence factors similar 

to those found in E. coli isolated from other colibacillosis lesions in 

poultry. Two hundred and thirty-seven birds from thirty broiler flocks 

were examined. Eighty two (34.6%) of 237 birds condemned for 

cellulites had gross lesions of colibacillosis and 18.9% of E.coli 

isolates from the 2 types of lesions belonged to the same 0-serogroup 

E.coli of serogroups O:78, O:1 and O:2 were predominated. 

El-Boraay and Abo-Table(2002) recovered a total of 82 isolates 

of E.coli from the livers, lung and intestine of the autosied 110 

broilers (74.54%) at kalyobia province. Serotyping of the 82 E.coli 

isolates were carried out and resulted in, O78:,O55:K60 and O26:K 

60. 

Taha (2002) recovered E.coli from 19 chickens samples in 

Sharkia province. E. coli isolates were 7123 with a frequency among 

other enteric bacteria 30.4% seven E.coli isolates were serotyped and 

resulted in (one each) O8:K25; O15:K74; O55:K59 (B5) and O126: 

K71 (B16) with a percentage of 14.3% each. 

Mc Peake et al., (2005) detected that a total of 114 avian 

pathogenic Escherichia coli (APEC) isolates were collected from 

cases of colisepticemia according in broilers (77) and layers (37) 

8


within Ireland. In addition, 45 strains isolated from faeces of healthy 

birds included for comparison. All isolates were serogrouped and 

examined for known virulence factors, mostly by PCR. The O78 

serogroup represented 55 and 27% pf broiler and layer colisepticaemic 

isolates respectively. 

Landman and Cornelissen (2006) stated that E. coli can induce 

peritonitis, a major cause of mortality in layer hens, but also other 

localized and systemic infections. E.coli infections have also been 

described in turkeys, geese and ducks and are thought to be a cause of 

significant economic losses. However, little is known about the real 

economic impact of the disease in layer chickens. Bacteriological 

analysis is required to establish a defined diagnosis because other 

pathogens can also cause salpengitis and peritonitis in layer hens. 

Antibiotics chosen on basis of sensitivity testing and other 

pharmacokinetic propertied can be used as therapy; however residues 

in eggs may occur. Auto-vaccines are often used as prevention 

because in practice effective protection is only achieved against 

homologous E. coli serogroups. 

2.1.2. Salmonella 

Barbour et al. (1983) examined a total of 412 feed samples and 

632 litter samples from 15 poultry farms (2 breeding farms and 13 

rearing farms) for detecting salmonella species in Saudi Arabia 

poultry farms. Twelve of these farms had salmonella species in litter, 

five farms had salmonella species in feed and four had salmonella 

species in broth feed and litter. Seventeen feed samples (4.13%) and 

121 litter samples (19.15%) were contaminated with salmonella 

9


species. Sixteen salmonella serotypes were encountered, of which 6 

were found in broth feed and litter. The five most frequently isolated 

salmonella serotypes in feed and litter were S. concord, S. colen, S. 

livingstone, S. manhattan and S. paratyphi. 

Shahata (1983) examined 240 dead chicken from upper Egypt 

poultry farms for recovery of salmonella species. Nine different 

salmonella serotypes from 20 isolates with a rcovery rate 8.33% were 

obtained, S. typhimunium was one of the identified serotypes. 

Likov et al. (1984) studied the index of infection in two broiler 

farms from the period of hatching up to the end of fattening. The fecal 

samples revealed high index of infection (15 to 31.7%) among 

revealed high index of infection (15 to 31.7%). Among the clinically 

normal birds, sixteen, 26-day old, salmonella free birds were divided 

into 2-groups and kept together with 2 birds (one in each group) orally 

infected with Salmonella isagni. Up to the 13 th day, the percentage of 

infected bird contacts reach to 92.3% and 100% in two groups. This 

made it responsible to believe that there was a chain of infections 

following one after another among healthy birds, maintained by 

sources of Salmonella species. 

Soerjadi Liem and Cumming (1984) determined the incidence 

of Salmonella species in 20 broiler flocks at time of slaughter in 

Australia. The Salmonella species incidence was tested by culturing 

the ceca of 50 randomly selected chickens per flocks. The results 

demonstrated high level of carriers (more than 30%) in 6 flocks, a low 

level of carriers (1 to 10%) in 2 flocks and 8 flocks from, which no 

10


salmonella species were isolated. Five salmonella serotypes were 

identified; S. typhomurium was the most common one of them. 

Irwin et al. (1989) studied the prevalent of Salmonella species in 

Ontario broiler chicken by culturing cloacal samples from 500 

individual birds selected from 50 poultry farms. Results of cloacal 

samples revealed, the presence of 19/500 (3.8%) samples contained 

salmonella species. Nine different salmonella species serotypes were 

isolated, the most common being S.hadar, S.heidelberg and S. 

mbandaka. 

Machado and Bernardo (1990) carried out a bacteriological 

examination of 300 chicken carcasses during the period of 1986-1987. 

it was found that 57% of the examined carcasses were contaminated 

with salmonella species. By serotyping, 3% of the salmonella strains 

were S.typhimurium. 

Poppe et al. (1991) estimated that the prevalence of salmonella 

species among Canadian commercial broiler flocks. They found that 

environmental (Litter and/or water) samples from 226 of 294 (76.9%) 

were contaminated with salmonella litter samples were more often 

contaminated with salmonella than water samples (47.4 versus 

12.3%). The most prevalent salmonella serovars from 50 salmonella 

strains were S.hadar, S.infantis and S.schwarzengrund; they were 

isolated from samples of 98/294 (33.3%), 26/294 (8.8%) and 21/294 

(7.1%) respectively. Thirty nine from 290 (13.4%) feed samples were 

contaminated with Salmonella enteritidis was isolated from 

environmental samples of 9/294 (3.1%). 

11


Osman (1992) collected 150 random, samples from different 

broiler farms at Sharkia Government she found that 45 stains were 

positive for salmonella species with an incidence of 30%. Serological 

typing of the isolated salmonella species revealing 21 (46.7%) were S. 

pullorum, 9 (20%) were S.gallinarum, 7 (15.6%) were S.typhimurium 

and 8/17 were not identified. 

Draz et al. (1996) examined 60 water and 45 litter samples from 

poultry environment and farms. Salmonella species was isolated with 

the frequency of 1.7% (1/60) and 2.2% (1/45) respectively. The 

isolated water borne strain was identified as S. typhimurium. 

Rusul et al. (1996) estimated the prevalence of salmonella 

species among broilers retailed at wet markets and processing plants 

in Malaysia. A total of 158 out of 445 (35.5%) and 52 out of 104 

(50%) broiler carcasses obtained from wet markets and processing 

plants were contaminated with Salmonella species respectively. 

Salmonella species was isolated from 14 out of 98 (14.3%) samples of 

intestinal content. Litter sample from broiler and breeder farms were 

positive for Salmonella species with the frequency of 8/40 (20%) and 

2/10 (20%), respectively. Examined breeder, broilers and layers 

flocks, Salmonella species were recovered from litter sample (42%), 

water in drinking troughs (36%), feed left over in feed trays (28%), 

water in the main tanks (17%), cloacal swabs (13%) and stock feed 

(8%). 

12


Sasipreeyajan et al. (1996) detected Salmonella species in 30 

broiler flocks in Thialand from October 1991 to August 1992. 

Hoop and Albicker-Rippinger (1997) isolated 37 Salmonella 

gallinarum pullorum strains from dead poultry between 1986 and 

1996. All strains expect one belonged to the biovar pullorum. 

Boonmar et al. (1998) assessed the prevalence of Salmonella 

species in chickens in Thailand. In 1997, 22 serovars of Salmonella 

species were isolated from 72 out of 100 chicken meat samples 

purchased from 10 retail markets in Bangkok and 20 out of 200 

chicken sample from one slaughterhouse and 19 out of 285 chicken 

feces obtained from three farms located in the east region of Thailand. 

The most predominant serovars was S.enteritidis, which was isolated 

from 28% of the most retail chicken feces samples examine. 

El-Morsi (1998) studied the incidence of the isolated Salmonella 

in the examined 25 liver samples of poultry. Positive samples for 

Salmonella species were 3 strains (12%). Serological typing for the 3 

isolated strains revealing one strain was S.typhimurium (33.3%) and 2 

strains were S. gallinarum (66.6%). 

Al-Nakhli et al. (1999) described the source and prevalence of 

pathogenic salmonella serovars among poultry forms in Saudia Arabia 

at the period of 1988 till 1997. A total of 1052 poultry samples and its 

environment were examined from, which Salmonella species were 

recovered with a percentage of 4%. Eleven salmonella serogroups 

representing 38 different salmonella serovars were identified. The 

13


majority of 276 isolates (26.2%) of salmonella species were recovered 

from heart, liver and intestine of the broilers and layers. S.entertidis 

(85 isolates, 98.8%), S.virchow (48 isolated, 57.8%) S. paratyphi (41 

isolates, 57.71%) and S.infants (30 isolates, 20.6%) were distributed in 

poultry and poultry environment. 

Mohammed et al. (1999) collected 200 faecal samples from 

living chickens in addition to 180 samples collected from chicken 

environment, Litter (75), feed (30) and water (75) at Kafr-El-Sheikh 

province. The total incidence of salmonella was 2.5% in chicken, 

5.33% in Litter, 3.33% in feed and 2.66% in water. The number of 

isolated S.entertidis strains from litter was (1) and feed (1), mean 

while S.typhimurium was isolated from chicken (2), S.anatum were 

isolated from chicken (3) and S.pullorun were isolated from chicken 

and water (1 each in number). 

Hatab (2001) collected 25 poultry feed samples from poultry 

houses at different localities in Dakahila province. Salmonella species 

were isolated from 3 raw mash samples with an incidence of 12%. 

Sertoyping of the three isolated Salmonella species revealed that 2 

strains were S. typhimurium and one untypable strain. 

Mohamad (2002) illustrated the occurrence of Salmonella 

species in 31 poultry manure samples in federal Germany with a 

frequency of 2/31 (6%). Both isolates were serotyped as S.agona. 

Salmonella species was counted in poultry manure with mean number 

of 162 cell per gram manure. 

14


Moreno et al. (2007) mentioned that antimicrobial resistance is 

an increasing phenomenon but its quantitative estimation remains 

controversial. The classical resistance percentage approach is not well 

studied to detect either emergence or low levels resistance. They 

performed E.coli enumeration in facial samples of broilers (82 pooled 

samples). Antimicrobial susceptibility of isolates was supplemented 

with 1 µg/ml of cefotaxime for E. coli detection, 93% (76/82) of 

broiler pooled samples tested positive. 

2.1.3. Frequent Enterobacteriaceae in broiler and surrounding 

environment. 

Verma and Adlakha (1971) stated that, out of 359 chickens 

suffered from pneumonia, sepcticemia, egg peritonitis, enteritis and 

persistent yolk sac. Klebsiella species were isolated from 12 chickens. 

Proteus species were isolated from 4 and paracolon bacteria from one. 

Karim and Ali (1976) examined 200 dead chick embryos of 22 

days old. Proteus species were isolated from dead chickens embryos. 

Sarakbi (1979) reported the incidence of Klebsiella species in 

different organs of ill and dead one day-old chicks. A total of 438 

samples from yolk-sac, heart blood, liver (146 each) were examined 

for the incidence of Klebsiella species, and result in recovery rates of 

17 (11.64), 10 (6.85%) and 10 (6.85%), respectively. K. pneumoniae 

was identified in 10, 7, 7 strains recovered from yolk sac, heart blood 

and liver samples respectively while K.ozoenae was isolated from two 

yolk sac samples. 

15


Abd-Allah (1981) mentioned that 275 chicks died in the first 10 

days yielded Klebsiella species from heart blood, lung and unabsorbed 

yolk sac. 

Brenner (1984) found that members of Citrobacter species are 

opportunistic pathogens, also Klebsiella species and enterobacterial 

species are opportunistic organisms. 

Morley and Thomson (1984) reported an outbreak of ocular 

disease caused by Klebsiella species affected a flock of 4 weeks-old. 

Orajaka and Mohan (1985) reported that Klebsiella species 

could cause embryo mortalities and excess losses in young chickens. 

Rosenberger et al. (1985) reported that Klebsiella species are 

environmental contaminants that occasionally cause embryo 

mortalities and excess losses in young chicken. 

Abdel-Gawad (1989) recovered Proteus mirabilis from 33 

(10.6%) out of 310 dead baby chicks. P. mirabilis strains were 

pathogenic to chicks, causing congestion of liver, heart, lung, spleen, 

kidney, unabsorbed yolk sac and omphalitis wither 24 hour after 

infection. 

Lin et al. (1993) found that Proteus morganis (Morganella 

morgani) cause 50% mortality in experimentally inoculated 4-weeks 

old chickens. 

16


Mohamad (1996) examined 150 baby chicks; disease and freshly 

dead at Sharkia Governorate broiler farms. He found the incidence of 

Klebsiella species (33.3%), Proteus species (22%) and Psendomonas 

aeroginosa (8.7%). 

Barnes and Gross (1997) reported that Proteus morgani has 

been associated with respiratory disease in chickens. 

Sakr (1998) found that 15 Salmonella species (2.4%), 14 

Klebsiella species (2.4%), 12 Proteus species (2.1%), 9 Shigella 

species (1.5%), 7 Citrobacter species (1.2%) and 8 Pseudomonas 

aeruginosa (1.3%). Serological identification of the isolated forty E. 

coli strains revealed 9 O:128 (20%), 7 O:1 (15.6%), 3 O:14 (13.3%) 

5 O:26 (11.1%), 4 O:1 (8.9%), 4 O:125 (8.9%) and 3 O:3 with 

prevalence of 6.7%. 

Hatab (2001) reported that mash ration of broilers proved to be 

highly contaminated with many pathogenic and potentially pathogenic 

organisms of Enterobacterialecie than pelleted feed Klebsiella species 

were isolated (32%) from raw mash, P. margani (M. morganii) (16%). 

Proteus mirabilis (8%), Salmonella species (12%), Enterobacterial 

species (16%) and Citrobacter species (8%), while in pellet feed 

Klebsiella species (24%), Proteus mirabilis (4%) and Citrobacter 

species (4%). 

Taha (2002) isolated Proteus species (13%), Klebsiella (4.3%), 

Citrobacter species (21.7%), Entrobacter species (4.3%) and 

Edwardsiella species (13%) from chicken in Sharkia. Enteric bacteria 

were isolated from water samples as Proteus (11.5%), Klebsiella 

17


species (11.5%), Citrobacter species (46.2%) and Enterobacter species 

(7.7%). 

Zaki et al (2002) collected 90 water samples from broiler farms 

(30 each). Coliforms were counted in reservoir tanks and drinkers as, 

97.53 and 4017.5 cells/100 ml water respectively Proteus, 

Pseudomonas, Klebsiella, Entrobacter and Citrobacter species were 

identified in reservoir tanks as, 11 (8.27%), 10 (7.52%), 13 (9.77%), 

13 (9.77%) and 8 (6.02%) respectively, meanwhile, in drinking water 

as, 14 (8.48%), 10 (6.06%), 18 (10.9%), 20 (12.12%) and 9 (5.45%), 

respectively. 

2.2. Antimicrobial sensitivity pattern for E. Coli 

El-Bakry et al. (1983) carried out sensitivity test on 30 isolates 

of E.coli (isolated from dead baby chicks) against furazolidone, 

gentamicin, tetracycline and chloramphenical. The ratio of sensitive 

strains to the totally tested ones were 80% (furazolidone), followed by 

70% (neomycin), 63% (gentamycin), 40% (choramphenicol) and 20% 

(tetracyclin). 

Giurov (1985) tested the susceptibility of 223 strains of E.coli to 

therapeutic agents with disk diffusion method. The organisms were 

Isolated from internal organs of birds died with colispticaemia. 

Serotyping revealed that O:1, O:2, O:4, O:8, O:26, O:78, O:111, 

O:103, O:141 and un-typable serotypes were 41, 70, 2, 3, 1, 70, 2, 1, 1 

and 32 in number resepectively. Highest number of strains proved 

sensitive to colistin (96.06%), the remaining drugs following in a 

descending order: flumequine (95.65%), amikacin (88.57%) car- 

18


pencillin (68.88%), furazolidone (83.13%) and kanamycin (61.67%) 

ampicillin (51.12%), chloramphenical (50.23%), and streptomycin 

(44.84%). 

Filali et al. (1988) isolated sixty two strains of E. coli (O:78, 

O:1&O:2) from 58 broiler farms suffering from respiratory signs and 

lesion characteristic to avian colibacillosis .All strains of isolated E. 

coli were sensitive to colistin, flumequine and gentamycin .A few 

strains were resistant to neomycin, nalidixic acid and trimethoprim. 

The frequently strains Resistant to nitrofurans, sulfonamides, 

chloramphenicol, spestinomycin and ampicillin was intermediate. 

Most strains were resistant to tetracycline. 

Adesiyum and Kaminjoli (1992) found that E. coli isolates were 

most resistant to streptomycin (81%) and tetracycline (79%) and least 

resistant to chloramphenicol (4%) and gentamycin (5%) The 

predominant resistance pattern for all isolate was streptomycintetracycline 

(28%). 

Osman (1992) reported that the sensitivity test for E. coli gave 

superior results with lincospectin (85-71%) followed by streptomycin, 

nitro furans, erythromycin, gentamycin, flumequine, nalidixic acid, 

sulfate with activity percentage of 80.9%, 78.5%, 73.8%, 6606%, 

59.5%52. 3%,42.8%, 40.4%, 23.8% and 19.5%, respectively. 

Meanwhile no effect could be observed with neomycin. 

Dinh and Nguyen (1995) studied the efficiency of antibiotics 

(streptomycin, chloramphenical, tetracycline, furazolidone, neomycin, 

19


ampicillin, sulphadimidine, trimethoprime sulphadimidine, combination 

of Kanamycin and gentamycin) on 201 enteropathogenic E. 

coli isolates. Streptomycin and tetracycline were the least effective but 

kanamycin and gentamycin were the most effective. The resistance 

pattern varied between the regions. 

Blanco et al. (1997) reported that ant microbial therapy is an 

important tool in reducing the enormous losses in portly industry 

caused by colibacillosis. Antimicrobial resistance testing of 468 avian 

E. coli strains isolated in Spain showed very high levels of resistance 

to trimethoprime-sulfamethoxazole (67%) and the flouroquinolones 

(13 to 24%). 

Abd-El-Mawla (1998) applied sensitivity test on 46 E. coli 

isolates versus to 11 different chemotherapeutic agents. It was noticed 

that all E.coli isolates were highly sensitive to neomycin, chloramphenicol, 

genataramycin and amikacin in a descending order of 

potently 93.6%, 91.3%, 89.1% and 82.6%, respectively. The tested 

isolates showed high resistance to penicillin G (100%), erythromycin 

(100%), trimethprime/sulphamethoxazole (80.4%) and flumequine 

(76.08%). A moderate sensitivity was encountered with tetracycline 

(73.90%), ampicillin (50%) and streptomycin (39.1%). 

El-Morsi (1998) found that E. coli showed a high resistance 

against ampicillin (100%), penicillin (97.67%) and tetracycline 

(69.77%). Other strains of E. coli showed high sensitivity to norfloxacin 

(93.02%), enrofloxacin (83.72%) and cefotaxim (72.09%). 

20


El-Ghamdi et al. (1999) compared antibiotic resistant E. coli 

isolates to ampicillin, chloramphenicol, getamicin, spectinomycin, 

tetracycline and trimethoprime spectinomycin, tetracycline and sulfamethoxazole 

ranged from 57% to 99.1% Although, amoxicillincluvalanate, 

ceftazidime And nitro furans ranged from zero to 2.6% 

resistance to spectonmycin reached 96% multi-drug resistance was 

alarmingly high. 

Lambie et al. (2000) isolated E. coli from diseased broilers. The 

isolates showed an increasing trend of resistance to amoxicillin, 

paramecia, gentamycin, nitrofurance, norfloxacin and sulphamethoxazole/ 

trimethoprim overall, norfloxacin appeared as the best 

antimicrobial drug for treatment. 

El-Sayed et al. (2001) stated that a high level of multi-resistant 

strains to the antibiotic. They found that 93.3% of the strains were 

ampicillin resistant and 73.3% of the strains were to amoxicillin and 

trimethoprim-sulfamethoxazol. 

2.3. Pathogenic Enterbacteria in Human 

2.3.1. E. coli 

Chapman et al. (2002) reported that phage therapy viruses that 

specifically target pathogenic. Bacteria has been developed over the 

last 80 years, primarily in the former soviet union, where it was used 

21


to prevent diarrhea caused by E. coli, among other things, in the red 

army, and was widely available over then counter. Presently phage 

therapy center in the republic of Georgia, However on January the 

2nd, 2007 the FDA gave approval to apply its O157: H7 killing phage 

in mist, spray or wash on live animals that will be slaughtered for 

human consumption 

Feng et al. (2002) stated that E. coli is one of the main species of 

bacteria living in the lower intestines of mammals, known as gut flora. 

When located in the large intestine. It actually assists with waste 

processing, vita mink production and food absorption. In 1885, it was 

discovered by the odor that E. coli strain O157:H9 is one of hundreds 

of strains of the bacterium that cause illness in human. E. coli are 

unable to sporulate so treatment with pasteurization or simple boiling, 

will kill all active bacteria. 

Alam and Zurek (2004) stated that if E.coli escape the intestinal 

tract through a perforation (hole or tear, for example from an ulcer 

ruptured appendix, or a surgical error) and enter the abdomen, they 

usually cause peritonitis that, E. coli are extremely sensitive to such 

antibiotics as streptomycin or gentamycin, so treatment with 

antibiotics is usually effective. This could rapidly change, E. coli 

rapidly aquired drug resistance. Certain strains of E. coli, such as E. 

coli O157:H7, E. coli O121 and E. coli O104:H21, are toxigenic 

(some produce a toxin very similar to that seen in dysentery). They 

can cause food poisoning usually associated with eating cheese and 

contaminated meat (contaminated during or shortly after slaughter or 

during storage or display). O157:H7 is farther notorious for causing 

22


serious ever life threatening completions like HUS (Hemolytic uremic 

syndrome). 

Szalanski et al. (2004) detected that E. coli can generally cause 

several intestinal and extra-intestinal infections such as urinary tract 

infections, meningitis, peritonitis, mastitis, septicemia and gramnegative 

peneumonia. The enteric E.coli are Gram-negative 

pneumonia. The enteric E.coli are divided on the basis of virulence 

properties into enterotoxigenic (ETEC), causative agents of diarrhea 

in humans, pigs, sheep, goats, cattle, dogs and horse, enteropthogenic 

(PEC) causative agent of diarrhea in human, rabbits, dogs and cats, 

enteroinvasive (ELEC), found only in human, enteroinvasive (ELEC, 

found only in human), verotoxigenic (VETC, found in pigs, cattle, 

dogs and cats), enterohaemorrhagic (EHEC, found in human, cattle 

and goats attacking porcine strains that colonize the gut in a manner 

similar to Human EPEC strains and enteroaggregative E.coli (Eagg 

E.c, found only in human). 

Sela et al. (2005) Although the urinary tract infection is more 

common in females due to the shorter urinary tract urinary tract 

infection is seen in both males and females, It is found in equal 

proportions in elderly the urinary tract through the urethra (as 

ascending infection), poor toilet habits can predispose to infection 

(doctors often device woman to wipe front to back, not back to front) 

but other factors are important (pregnancy in women, prostate 

enlargement in men and in many cases the initiating event is nuclear 

while ascending infections are generally the rule for lower urinary 

tract infections and cystitis the same may not necessarily hold for 

23


upper urinary tract like pyelonephritis which may be hematogenous in 

origin. Most cases of lower urinary tract infections in females are 

benign and do not need exhaustive laboratory work-ups. However, 

VTI in young infants must receive some imaging study, typically, a 

retrograde urethrogram, to ascertain the presence/absence of 

congenital urinary tract anomalies. Males too must be investigated 

further methods of X-ray, MR and CAT scan technology. 

Ahmed et al. (2006) mentioned that E. coli vaccines have been 

under development for many years. In March of 2006, a vaccine 

eliciting an immune response against the E. coli O157:H7 O-specific 

polysaccharide conjugated to recombinant exotoxin of Pseudomonas 

aeruginosa (O157-rEPA) was reported to be safe and immunogenic in 

children two to five years old. It has already been proven safe and 

immunogenic in adults. Phase iii clinical trial to verify the large-scale 

efficacy of the treatment is planned. In January 2007 the Canadian 

bio-pharmaceutical company Bioniche announced it has developed a 

cattle vaccine, which reduces the number of bacteria, shed in manure 

by a factor of 1000, to about 1000 bacteria per gram of manure. 

Girard et al. (2006) reported that a strain of E. coli is a group 

with some particular characteristics that make it distinguishable form 

other E. coli strains. These different are often detectable. Only on the 

molecular level; however, they may result in changes to the 

physiology or life cycle of the bacterium, for example leading to 

pathogenicity. Different strains of E. coli live in Different kinds of 

animals so it is possible to tell whether fecal material in water came 

from humans or from birds for example new strains of E. coli arise all 

24


the time from the natural biological process of mutation, and some of 

those strains have characteristics that can be harmful to a host animal. 

Although In most healthy adults humans such a strain would probably 

cause no more than about of diarrhea, and might produce no 

symptoms at all, in young children, people who are or have recently 

been sick, or in people taking certain Medications, an unfamiliar strain 

can cause serious illness and even death. Particular virulent example 

of such a strain of E. coli is E. coli O157:H7. 

Johnson et al. (2006) found that E. coli and related bacteria possess 

the ability to transfer DNA via bacteria conjugation, which allows anew 

mutation to spread through an existing population. It is believed that this 

process led to the spread of toxin extended-spectrum Beta-lactamase 

(ESBL)-producing E.coli and antibiotic-resistant strains of E.coli Are 

antibiotic-resistant strains of E.coli. ESBL- producing strains are bacteria 

that producing strains are bacteria that produce an enzyme called extendedspectrum 

beta lactamase, which makes them more resistant to antibiotics 

and makes the infections harder to treat. In many instances, only two oral 

antibiotics and every limited group intravenous antibiotics remain effective. 

Cheristie and Tim (2007) Appropriate Treatment depends on the 

disease and should be guided by laboratory analysis of the antibiotic 

sensitivities of the infecting strain of E.coli. As Gram-negative organism, 

E.coli are resistant to many antibiotics which are effective against Grampositive 

organism. Antibiotics which may be used to treat E. coli infection 

include (but are not limited to) amoxicillin as well as other semi-synthetic 

penicillin, many cephalosprians, carbapenems, aztreonam, trimelthoprimesulfamethoxazol 

ciprofloxacin, nitrofurantion and aminoglycosiders, Not 

all antibiotics are suitable for every disease caused by E.coli and the advice 

of a physician should be sought. Antibiotic resistance is a growing problem 

25


Some of this is due to overuse of antibiotics in human, but some of it is 

probably due to the use of antibiotics as growth promoters in food animals 

resistance to beta-lactam antibiotic has become more serious in recent 

decades as strains producing extended-spectrum beta-lactamases render 

many, if not all, of the penicillins and cephalosporins ineffective as 

therapy. Susceptibility testing should guide treatment in all infections. In 

which organism can be isolated for culture. 

Pearson (2007) recorded the presence of coliform bacteria in 

surface water is commonly used as model organism for bacteria in 

general. This is usually done using the MPN (most probable number) 

tests, this is usually probabilistic test which assumes bacteria meeting 

certain growth and biochemical criteria as E. coli and quantities it by 

various methods. Presence of E.coli numbers beyond certain cut-off 

indicates fecal contamination of water and indicates further 

investigation into the matter. E.coli is used for detection because there 

are a lot more conifers in human feces than there are pathogens 

(Salmonella typhi an example of such pathogen, causing typhoid 

fever) and E. coli is usually harmless, so it can’t get loose in the lab 

and hurt any one. However, sometimes it can be misleading to use E. 

coli alone as an indicator of human fecal contamination because there 

are other environments in which E. coli grows well, such as paper 

mills. 

2.3.2. Salmonella 

Martin et al. (2003) resistance of Salmonella and E. coli to 

extended-spectrum cephalosporins (ESCs) is being reported with 

increasing frequency. Ceftiofur, a veterinary ESC, may be used more 

often for the treatment of bacterial infections in animals. S. newport 

26


isolates from humans and animals are increasingly resistant to the 

ESCs. In humans, infections with ESC resistant Salmonella threaten 

the efficacy of ceftriaxone, the drug of choice for treating 

salmonellosis in children. They examined Salmonella isolates for 

susceptibility to 23 antimicrobials. Isolates resistant to ampicillin and 

amoxicillin/clavulanic acid that were additionally resistant to 3rd 

generation cephalosporins and/or a cephamycin were further 

characterized by several methods. We assessed plasmid profiles and 

PFGE patterns, used PCR and sequencing to determine the presence 

of the cmy-2 and other drug resistance genes, determined the 

isoelectric point of the beta-lactamases produced, and carried out 

conjugation, transformation and hybridization studies. E. coli isolated 

during other projects were also examined. Their results revealed that 

examination of all (119) S. newport isolates among 36,841 Salmonella 

isolates from 1993-2002 showed that 0.1% of the 1993-98 strains but 

0.4-0.6% of the 1999-02 strains were S. Newport. More than 50% of 

the strains from 1993-2002 were of bovine origin. None of the S. 

newport strains isolated before 2000 were multiply resistant to 

antimicrobials, whilst 32 of 33 bovine and 3 of 14 poultry S. newport 

strains isolated during the 2000-02 period were resistant to more than 

5 antimicrobials including the ESCs. All resistant strains possessed the 

cmy-2 gene. We also found cmy-2 encoded resistance to the ESCs 

among other Salmonella serovars and E. coli isolates. They concluded 

that the resistance of Salmonella and E. coli isolated from animals, the 

animal environment, and foods of animal origin to ESCs is increasing. 

This will limit treatment options when humans become infected with 

such highly drug resistant strains. 

27

28 


Dobrgan et al. (2006) reported that Salmonella enterica serovars 

typhi, paratyphi A, and sendai are human-adapted pathogens that 

cause typhoid (enteric) fever. The acute prevalence in some global 

regions and the disease severity of typhoid salmonella have 

necessitated the development of rapid and specific detection tests. 

Most of methodologies currently used to detect serovar typhi don’t 

identify serovars paratyphi A or sendai. To assist in this aim, 

comparative sequence analyses were performed at the loci of core 

bacterial genetic determinants and salmonella pathogenecity island 2 

genes encoded by clinically significant S. enterica serovars. Genetic 

polymorphisms specific for serovars typhi (at tarps), as well as 

polymorphisms unique to human-adapted typhoidal serovars (at 

second step), were observed. Further more, entire coding sequence 

unique to human-adapted Typhoidal salmonella strains (i.e. serovar 

specific genetic loci rather than polymorphisms) were observed in 

publicly available comparative genomic DNA micro array data sets.

4.Results 

A total of 295 chicken samples of liver , spleen and heart 

from each bird, in addition to 53 human urine specimens 

were subjected to bacterial isolation (table 5), only 245 

bacterial isolates out of 295 chicken specimens were 

recoverd from samples of one day old chicks, broiler and 

layers of Dakahlia province. 

Only 39 bacterial isolates were recovered from 53 human 

urine specimens from private laboratories(table 3). 

Table (3): Prevalence rate of isolated Enterobacteria from 

different sources 

Source 

Chicken 

specimens 

Human 

specimens 

Samples 

Liver and 

Spleen 

(295) 

Female urine 

(18) 

Male urine 

(30) 

Children urine 

(5) 

Recovered 

isolates/total 

samples 

245/295 

83.05% 

14/18 

77.7% 

22/30 

73.33% 

3/5 

60% 

45 

Recovered 

L.F/total 

samples 

205/295 

69.49% 

13/18 

27.2% 

18/30 

60.0% 

3/5 

60% 

Recovered 

NLF/ total 

samples 

40/295 

13.55% 

1/18 

5.5% 

4/30 

13.33% 

0.0 

0.0%

L.F=lactose fermenter NLF=Non lactose fermenter 

4.1-Enterobacteria in Chicken 

specimens. 

Out a total of 295 samples obtained from livers and 

spleen of broilers, one day old chicks and layers, 245 

samples were recovered as Enterobacterial isolates with 

prevalence rate 83.05% of total number. 205 out of 

295(69.49%) isolates were lactose fermenter isolates (table 

3, fig.1). Fourty isolates were recovered as non-lactose 

fermenter isolates with prevalence rate 13.55% of total 

isolates. 

One hundred and twenty five samples collected from 

broiler and 170 samples collected from both one day old 

chicks and layers. (4,5). Only 107 isolates from a total of 

125(85.6%) samples from broiler were recovered. 

77/125(61.6%) as lactose fermenter, meanwhile 

30/125(24%) were recovered as non lactose fermenter ( 

table 4). 

Concerning specimens from layers and one day old 

chicks, Incidence of obtained isolates as lactose fermenter 

and non lactose fermenter from each farm ( table 5 ) 

revealed that only 138 isolates from a total of 

170(81.1%)samples from broiler were recovered. 

128/170(75.2%) were lactose fermenter, meanwhile 

10/170(5.88%) were recovered as non lactose fermenter. 

46

Table (4): Incidence of chicken Enterobacteria obtained on 

MacConkeyۥs medium from broiler. 

Farm 

Bela 

Bela 

Total 

no. 

of 

samples 

% of obtained 

isolates grown 

On 

MacConkey/ 

Total samples 

% of LF Gr-ve 

isolates/ 

Total samples 

% of NLF 

Gr-ve/ 

Total 

samples 

isolates 

8 6/8(75%) 6/6(100%) 0.0 

Bian 6 5/6(83.3%) 5/5(100%) 0.0 

Mansoura 23 21/23(91.3%) 11/23(47.8%) 10(43.4%) 

Shahen 7 5/7(71.4%) 5/5(100%) 0.0 

Kalig 16 15/16(93.7%) 15/15(100%) 0.0 

Omda 18 15/18(83.3%) 10/18(55.5%) 5/18(27.7%) 

Asher2 11 10/11(90.9%) 5/11(45.4%) 5/11(45.4%) 

Asher3 22 20/22(90.9%) 15/22(68.1%) 5/22(22.7%) 

Atef-Abd 

-El Aziz 

9 5/9(55.5%) 5/5(100%) 0.0 

47

Setawy 5 5/5(100%) 0.0 5/5(100%) 

total 125 107/125(85.6%) 77/125(61.6%) 30/125(24%) 


Table (5): Incidence of chicken Enterobacteria obtained on 

MacConkeyۥs medium from one day old chicks and layers. 

Farm 

Total 

no. 

of 

sample 

s 

% of obtained 

isolates grown 

On 

MacConkey/ 

Total samples 

% of LF Gr-ve 

isolates/ 

Total samples 

% of NLF 

isolates/ 

Total 

samples 

Mansoura2 17 15/17(88.2%) 15/15(100%) 0.0 

Bian2 14 12/14(85.7%) 12/12(100%) 0.0 

Kalig2 21 10/21(47.6%) 10/10(100%) 0.0 

Omda2 15 10/15(66.6%) 10/10(100%) 0.0 

Asher 16 16/16(100%) 16/16(100%) 0.0 

Asher4 16 15/16(93.7%) 5/16(31.2%) 10/16(62.5%) 

Abohegaz 16 15/16(93.7%) 15/15(100%) 0.0 

Abohegazy 13 10/13(76.9%) 10/10(100%) 0.0 

Abosoultan 17 15/17(88.2%) 15/15(100%) 0.0 

Abosoultan 

2 

14 10/14(71.4%) 10/10(100%) 0.0 

48

Abolila 11 10/11(90.9%) 10/10(100%) 0.0 

Total 170 138/170(81.1 

%) 

128/170(75.2 


49 

%) 

10/170(5.88 

%)

Fig. (1)Tube lactose fermentation using phenol red 

indicator(yellow in positive test but faint red for control 

medium). 

4.2- Enterobacteria in Human urine: 

Eighteen, 30 and 5 urine samples from female urine 

male and children contained 14/18(77.7%), 22/30 (73.33%) 

and 3/5(60%) isolates respectively (table 3). Concerning 

lactose fermenter isolates, the samples contained 13/18 

(72.2%), 18/30 (60.0%) and 3/5(60%) isolates 

respectively(table 3). 

Concerning non lactose fermenter isolates, the samples 

contained 1/18(5.5%), 4/30(13.33%) and 0.0(0.0%) isolates 

respectively(table 3). 

4.3-Biotyping of chicken Enterobacteria. 

Biochemical tests including, growth on MaCconkeyۥs 

medium oxidation fermentation test (O-F), growth on triple 

sugar iron agar (TSI, fig.3), Xylose Lysine decarboxylase 

(XLD.fig.2), Indol, M.R, V.P, Citrate(fig.4), urease, sugar 

50

fermentation (Lactose, Maltose, Mannitol, Sucrose) revealed 

that: 

Lactose fermenter isolates from broiler (table 6) were 

E.coli and Citrobacter diversus among tested isolates and 

represented 68/107 (63.55%) and 9/107 (8.41%) respectively. 

Meanwhile non Lactose fermenter isolates were salmonella, 

pseudomonas and proteus and represented 24/107(22.42%), 

4/107(3.7%), 2/107(1.86%) respectively 

Table(6)Incidence isolates after identification according to 

biochemical reactions. 

Isolate type 

E.coli 

Salmonella 

Broiler 

Chickens 

Layers and One 

day old chicks 

68/107 105/138 

(63.55%)* (76.08%)* 

24/107 

(22.42%) 

7/138 

(5.07%) 

51 

females 

12/13 

(92.30%)* 

1/1 

(100%) 

males 

human 

16/18 

(88.88%)* 

3/4 

(75%) 

children 

3/3 

(100%)* 

0.0

Pseudomona 

s spp. 

Citrobacter 

diversus 

4/107 

(3.7%) 

9/107 

(8.41%) 

Proteus spp 2/107 

(1.86%) 

2/138 

(1.44%) 

23/138 

(16.6%) 

1/138 

(0.72%) 

0.0 

1/13 

(7.69%)* 

1/4 

(25%) 

2/18 

(11.11%) 

0.0 

0.0 

0.0 0.0 0.0 

*Percentage to total number of isolates obtained on MacConkey 

medium 

Lactose fermenter isolates from one day old chicks 

and layers (table 6) were E.coli and citrobacter diversus 

among tested isolates. They represented 105/138(76.08%) 

and 23/138(16.6%) respectively. Meanwhile non Lactose 

fermenter isolates were salmonella, pseudomonas and 

proteus and represented 7/138 (5.07%), 2/138 (1.44%) 

and1/138 (0.72%) respectively 

4.4-Biotyping of urine isolates 

Lactose fermenter isolates from females urine 

subjects(table 6, Fig.1) were E.coli and citrobacter diversus 

among tested isolates and represented 12/13 (92.30%) and 

1/13(7.69%) respectively. Mean-while non Lactose fermenter 

52

isolates were salmonella, pseudomonas and proteus and 

represented 1/1(100%),0.0 and 0.0 respectively. 

Lactose fermenter isolates from males urine 

subjects(table 6) were E.coli and Citrobacter diversus among 

tested isolates and represented 16/18 (88.88%) and 2/18 

(11.11%) respectively. Meanwhile non Lactose fermenter 

isolates were salmonella, pseudomonas and proteus and 

represented 3/4 (75%),1/4(25%) and 0.0 respectively. 

Lactose fermenter isolates from children urine 

subjects(table 6) were E.coli and Citrobacter diversus among 

tested isolates and represented 3/3(100%) , 0.0 and 0.0 

respectively. Meanwhile non Lactose fermenter isolates were 

salmonella, pseudomonas and proteus and represented 0.0, 

0.0 and 0.0 respectively 

53

Fig.(2)Xylose lysine decarboxylase(XLD) medium, only in the 

fourth tube (right side) the isolate fermented sugars producing 

yellow colour, no H2s was produced. 

54

Fig.(3)Triple sugar iron agar(TSI), third and fourth tubes(right 

side),Acid slant(yellow) over Acid butt(yellow) no H2S. with 

gas in the third tube. First and second are control. 

4.5-Pathogenicity test. 

Two weeks broiler inoculated with 5.8X10 5 fresh 

subcultured E.coli isolates, after 2days depression, was 

observed, P.M. revealed congesion of internal organs (Fig.5), 

liver specimens were cultured for re-isolation of the same 

inoculated microorganisms. 

4.6-Results of serotyping of E.coli isolates. 

55

Dignostic E.coli polyvalent O antisera revealed O1, 

O135 , O157 and O78 serotypes in some broiler , layers , and 

one day old chicks. 

4.7-Results of antibiogram on isolates from one day old 

chicks: 

Testing chemotherapeutic agents including Cefotaxime, 

Ciprofloxacin, Amoxycyclin, Gentamycin, (Sulpamethoxazole, 

Trimethoprim), Colistin, Erythromycin, Rifmpicin, Streptomycin 

and Doxycycline against biotyped isolates (table 7) showed 

that:- 

Only the isolate from Asher1 farm was sensitive to all 

drugs, Asher3 farm isolate was sensitive to all drugs except 

(Sulpa-methoxazole, Trimethoprim), Abohegazy farm isolate 

was sensitive to all drugs except to (Sulpamethoxazole, 

Trimethoprim), Asher4 farm isolate was resistant to seven 

drugs and sensitive only to three drugs (Cefotaxime, 

Ciprofloxacin and Colistin). 

The results also showed that all the isolates were 

sensitive to Cefotaxime and Ciprofloxacin also all farms 

isolates were sensitive to colistin except the isolate from 

Mansoura1 farm. 

56

Fig.(4).Citrate negative E.coli isolate. 

57

Fig.(5)Pathogenicity lesions showing congested liver and 

pericharditis after challenge with 5.8X10 5 fresh E.coli culture. 

58

Fig.(6)An isolate was sensitive to gentamycin, ciprofloxacin 

and nitrofurantoin. 

59

Fig.(7) An isolate was sensitive to ciprofloxacin, 

amikacin and gentamycin. 

Table (7) Antibogram of isolates from one day old chicks against ten 

chemotherapeutic agents. 

60

Farm 

Mansoura1 

Omda1 

Asher3 

Asher4 

Abohegazy 

Absoltana1 

Khalig 

Asher1 

Omda2 

CTX30 CIP Amox GN SXT25 CT DO E 

S 

S 

S 

S 

S 

S 

S 

S 

S 

Abosoltana2 S 

Mansoura2 

Abolila 

S 

S 

Abohegazy2 S 

Khalig2 

S 

S 

S 

S 

S 

S 

S 

S 

S 

S 

S 

S 

S 

S 

S 

R 

R 

S 

R 

R 

R 

R 

S 

R 

R 

R 

S 

S 

R 

S 

R 

S 

R 

S 

S 

S 

S 

S 

S 

S 

S 

S 

S 

R 

R 

R 

R 

S 

S 

R 

S 

R 

R 

R 

R 

R 

R 

R 

S 

S 

S 

S 

S 

S 

S 

S 

S 

S 

S 

S 

S 

S 

S 

S 

R 

S 

S 

S 

S 

S 

S 

R 

R 

S 

R 

R S 

R S 

S S 

R R 

S S 

R S 

R S 

S S 

S R 

S S 

R R 

S R 

S R 

S S 

RD ST 

R = Resistant S = Sensitive CTX30 = Cefotaxime CIP =Ciprofloxacin 

GN = 

Gentamycin SXT25 = Sulfaphmethaxazole/Trimethoprime CT = Colistin 

E = Erythromicin RD = Rifampicin ST = Streptomycin AMP = Ampicillin 

DO = Doxycycline 

4.8-Sensitivity pattern of the isolates from one day old 

chicks. 

Fourteen isolates from one day old chicks were tested 

10 chemo-therapeutic agents including Cefotaxime, 

61 

S 

S 

S 

R 

S 

S 

S 

S 

R 

R 

S 

S 

S 

R

Ciprofloxacin, Amoxicillin Gentamycin, (Sulpamethoxazole / 

Trimethoprim), Colistin, Doxycycline Erythromycin, Rifmpicin 

and Streptomycin ), sensitivity of the isolates to one or more 

drugs was recorded (table 8). 

Only one Salmonella isolate from Asher1 farm out of 14 

isolates 1/14 (7.1%) was sensitive to all examined drugs, 

there were no sensitive isolates to only one or only two or 

only four drugs, 1/14 (7.1%) Salmonella isolate from Asher4 

farm was sensitive to only three drugs, they were Cefotaxime, 

Ciprofloxacin and Colistin. 

One E.coli isolate from Mansoura2 farm was sensitive to 

only five drugs which were Cefotaxime, Ciprofloxacin, Colistin, 

Gentamycin and Streptomycin. 3 E.coli isolates from 

Mansoura1, Omda1 and Omda2 farms were sensitive to only 

six drugs, they were Cefotaxime, Ciprofloxacin, Gentamycin, 

Doxycycline, Rifmpicin and Streptomycin for Mansoura1 

isolates and Cefotaxime, Ciprofloxacin, Doxycycline, 

Rifmpicin, Streptomycin and Colistin for Omda1 and 

Cefotaxime, Ciprofloxacin, Gentamycin, Colistin, Doxycycline 

and Erythromycin for Omda2 meanwhile 4 E.coli isolates from 

Khalig, Abosoltana2, Abolila and Khalig2 were sensitive to 

only seven drugs, they were Cefotaxime, Ciprofloxacin, 

Gentamycin, Colistin, Doxycycline, Rifmpicin and 

Streptomycin for Khalig and Cefotaxime, Ciprofloxacin, 

Gentamycin, Colistin, Doxycycline, Rifmpicin and 

Erythromycin for Abosoltana2 and Cefotaxime, Ciprofloxacin, 

62

Gentamycin, Colistin, Erythromycin, Streptomycin and 

Amoxycylin for Abolila farm. 

Two E.coli isolates from Abosoltana1 and Abohegazy2 

were sensitive to only eight drugs which were Cefotaxime, 

Ciprofloxacin, Gentamycin, Colistin, Doxycycline, Rifmpicin, 

Streptomycin and (Sulpamethoxazole/Trimethoprim) for 

Abosoltana1 and Cefotaxime, Ciprofloxacin, Gentamycin, 

Colistin, Doxycycline, Erythromycin, Streptomycin and 

Amoxycylin for Abohegazy2. 

Two E.coli isolates from Asher3 and Abohegazy farms 

were sensitive to only nine drugs which were Cefotaxime, 

Ciprofloxacin, Gentamycin, Colistin, Doxycycline, 

Erythromycin, Streptomycin, Amoxycylin and Rifmpicin for 

Asher3 farm and Cefotaxime, Ciprofloxacin, Gentamycin, 

Colistin, Doxycycline, Erythromycin, Streptomycin, Rifmpicin 

and (Sulpamethox-azole, Trimethoprim) for Abohegazy farm. 

63

Table (8) Sensitivity pattern of the isolates from one day old 

chicks against 10 chemotherapeutic agents. 

Sensitivity of MicroMicro- 

Sensitivity 

organisms to one 

or more Drugs 

organisms incidence 

Sensitivity to all drugs Salmonella 1/14 (7.1%) 

Sensitivity to only one drug Nil 

Sensitivity to only two drugs Nil 

Sensitivity to only three drugs Salmonella 

Sensitivity to only four drugs Nil 

Sensitivity to only five drugs E.coli 

Sensitivity to only six drugs E.coli 

Sensitivity to only seven drugs E.coli 

Sensitivity to only eight drugs E.coli 

Sensitivity to only nine drugs E.coli 

0.0 

0.0 

1/14 (7.1%) 

0.0 

1/14 (7.1%) 

3/14 

(21.4%) 

4/14 

(28.5%) 

2/14 

(14.2%) 

2/14 

(14.2%) 

4.9-Results of antibiogram on layer isolates. 

Eleven isolates from layers were tested 11 

chemotherapeutic agents including Cefotaxime, Ciprofloxacin, 

Ampicillin, Spectam, Gentamycin, (Sulpamethoxazole/ 

64

Trimethoprim), Colistin, Erythromycin, Rifmpicin, Streptomycin 

and Doxycycline), the results revealed that:- 

Asher4 farm isolates (table 9) showed resistance to (Sulpha- 

methoxazole, Trimethoprim) and sensitivity to all other used 


The isolate from Asher1(A), the isolate from Asher1(B) 

and that from Bian2(B) farm were the most resistant isolate as 

they were resistant to six chemotherapeutic agents also the 

results recorded that the isolates from all farms were sensitive 

to Cefotaxime and Ciprofloxacin meanwhile the isolates from 

all farms were resistant to Colistin except the isolate from 

Abohegazy farm and isolate from Bian2(B). 

65

Table (9) Antibiogram of layer isolates against 11 

chemotherapeuticagents. 

Farm CTX30 CIP AMP GN SXT25 CT DO E RD ST SH 

Abosoltana S 

Abohegazy S 

Bian 2(A) S 

Asher1(A) S 

Asher3 S 

Mansoura1 S 

Mansoura2 S 

Asher1(B) S 

Bian 2(B) S 

Asher2 S 

Asher4 S 

S 

S 

S 

S 

S 

S 

S 

S 

S 

S 

S 

R 

R 

S 

R 

S 

S 

S 

S 

S 

R 

S 

S 

S 

S 

R 

R 

S 

R 

S 

R 

R 

S 

S 

S 

S 

R 

R 

S 

S 

R 

R 

S 

S 

R 

S 

R 

R 

R 

R 

R 

R 

S 

R 

R 

S 

S 

S 

S 

R 

R 

S 

R 

R 

R 

S 

S R 

S S 

S S 

S S 

S R 

S S 

S R 

S S 

R S 

R R 

S S 

R = Resistant S = Sensitive CTX30 = Cefotaxime CIP =Ciprofloxacin 

GN = 

Gentamycin SXT25 = Sulfaphmethaxazole/Trimethoprime SH = Spectam 

CT = Colistin E = Erythromicin RD = Rifampicin ST = Streptomycin 

AMP = Ampicillin DO = Doxycycline 

4.10-Sensitivity Pattern of layer isolates. 

Sensitivity Patterns of Eleven isolates from layers were 

recorded against 11 chemotherapeutic agents including 

Cefotaxime, Ciprofloxacin, Ampicillin, Spectam, Gentamycin, 

66 

S 

R 

R 

S 

S 

R 

R 

R 

S 

S 

S 

S 

S 

S 

R 

R 

R 

S 

R 

R 

R 

S

(Sulpamethox-azole/ Trimethoprim), Colistin, Erythromycin, 

Rifmpicin, Strepto-mycin and Doxycycline), the sensitivity 

results of the isolates(table 10 ) to only one or more drugs 

revealed that:- 

There were no isolates sensitive to all drugs also no 

isolates were sensitive to only one or only two or only three 

drugs. 1/11(9.09%) isolate from Asher2 farm was sensitive to 

only four drugs, they were Cefotaxime, Ciprofloxacin, 

Gentamycin, and Rifmpicin. 

1/11(9.09%) isolate from Asher2 farm was sensitive to 

only five drugs, they were Cefotaxime, Ciprofloxacin, 

Doxycycline, Rifmpicin and Ampicillin. 

3/11 (27.2%) E.coli isolates from Asher1(A), Asher1(B) 

and Bian2(B) farms were sensitive to only six drugs which 

were Cefotaxime, Ciprofloxacin, Colistin, Doxycycline, 

Erythromycin and Rifmpicin for Asher1(A) and Cefotaxime, 

Ciprofloxacin, Erythromycin, Rifmpicin and Ampicillin for 

Asher1(B) and Cefotaxime, Ciprofloxacin, Erythromycin, 

Ampicillin, Rifmpicin and (Sulpamethoxazole, Trimethoprim) 

for Bian2(B) isolate. 

2/11(18.18%) E.coli isolate from Mansoura1 and 

Mansoura2 farms were sensitive to only seven drugs, they 

were Cefotaxime, Ciprofloxacin, Erythromycin, Ampicillin, 

Strepto-mycin, Gentamycin and Doxycycline for Mansoura1 

67

isolate and Cefotaxime, Ciprofloxacin, Ampicillin, Gentamycin, 

Doxycycline, colistin and Spectam for Mansoura2 isolate. 

1/11(9.09%) E.coli isolate from Abosoltana farm was 

sensitive to only eight drugs, they were Cefotaxime, 

Ciprofloxacin, Gentamycin, Rifmpicin, colistin, spectam, and 

Doxycycline. 

2/11(18.18%) E.coli isolate from Abohegazy and 

Bian2(A) farms were sensitive to nine drugs which were 

Cefotaxime, Ciprofloxacin, Colistin, Doycyclin, Gentamycin, 

Erythromycin, Streptomycin, (Sulphamethazole, Trimethoprim) 

and Spectam for Abo-hegazy isolate and Cefotaxime, 

Ciprofloxacin, Colistin, Doycyclin, Gentamycin, Erythromycin, 

Streptomycin, Spectam and Ampcillin for Bian2(A) isolate 

farm. 

1/11(9.09%) E.coli isolate from Asher4 farm was 

sensitive to ten drugs which were Cefotaxime, Ciprofloxacin, 

Colistin, Doycyclin, Gentamycin, Erythromycin, Streptomycin, 

Spectam, Ampcillin and Rifmpicin. 

68

Table(10)Sensitivity pattern for isolates from layers. 

Sensitivity of 

Microorganisms to one or 

more Drug 

Type of 

Microorganisms 

Sensitivity to all drugs Nil 

Sensitivity to only one drug Nil 

Sensitivity to only two drugs Nil 

Sensitivity to only three 

drugs 

Nil 

Sensitivity to only four drugs salmonella 

Percentage of 

Sensitivity 

0.0 

0.0 

0.0 

0.0 

1/11 (9.09%) 

Sensitivity to only five drugs salmonella 1/11(9.09%) 

Sensitivity to only six drugs E.coli 

Sensitivity to only seven 

E.coli 

drugs 

Sensitivity to only eight 

E.coli 

drugs 

Sensitivity to only nine drugs E.coli 

3/11 (27.2%) 

2/11 (18.1%) 

1/11 (9.09%) 

2/11 (18.1%) 

Sensitivity to only ten drugs E.coli 1/11 (9.09%) 

4.11-Results of antibiogram on isolates from broiler. 

Ten isolates from broiler farms were tested 11 

chemotherapeutic agents(table 11) including Cefotaxime 

(CTX30), Ciprofloxacin(CIP), Gentamycin(GN), 

69

Sulphamethazole, trimetho-prime(SXT25), Spectam(SH), 

Colistin(CT), Erythromycin(E), Rifampicin(RD), Streptomycin 

(ST), Ampicillin (AMP), Doxycyclin (DO) . 

Results showed that (table 14)only one isolate of 

Salmonella from Omda farm was sensitive to all the tested 

drugs, meanwhile E.coli isolates from Bian and Shahen farms 

were resistant to seven drugs. The results also recorded that 

all isolates were sensitive to Cefotaxime meanwhile there was 

multiple resistance to (Sulphamethazole, trimethoprime), 

Colistin and Spectam. 

Table(11)Antibiogram of isolates from broiler against 11 


Setawy Farm CTX30 S CIP S GN R SXT R25 

SH R CT R DO R SE RD R ST S AMRP Asher2 

Bella 

S 

S 

Bella 

Asher3 S 

Bian S 

Mansoura S 

Shahen S 

Khalig S 

Omda S 

Atef abd 

El-Aziz 

S 

S 

S 

S 

S 

S 

S 

R 

S 

S 

R 

S 

S 

R 

S 

R 

R 

S 

R 

R 

R 

R 

R 

R 

R 

R 

S 

R 

70 

R 

S 

R 

R 

R 

R 

R 

S 

R 

R 

S 

R 

R 

R 

R 

R 

S 

R 

S 

S 

R 

S 

R 

S 

R 

S 

S 

R S 

R S 

R S 

R R 

R R 

R S 

R S 

S S 

S S 

S 

R 

S 

R 

R 

S 

R 

S 

S 

S 

R 

R 

R 

R 

R 

R 

S 

R

R = Resistant S = Sensitive CTX30 = Cefotaxime CIP =Cipro-floxacin 

GN = Gentamycin SXT25 = Sulfaphmethaxazole , Trimethoprime SH = 

pectam CT = Colistin E = Erythromicin RD = Rifampicin ST = Streptomycin 

AMP = Ampicillin DO = Doxycycline 

4.12-Sensitivity pattern of broiler isolates. 

Sensitivity Patterns of ten isolates from broiler were 

recorded against 11 chemotherapeutic agents including 

Cefotaxime, Cipro-floxacin, Gentamycin, (Sulpamethox-azole/ 

Trimethoprim), Spectam, Colistin, Doxycycline, Erythromycin, 

Rifmpicin, Strepto-mycin Ampicillin, and), the sensitivity results 

of the isolates(table 11) to only one or more drugs revealed 

that 

1/10 (10%) Salmonella isolate was sensitive to all 

drugs but no isolates were sensitive to only one drug , 1/10 

(10%) E.coli isolate from khalig farm was sensitive to only two 

drugs which were Cefotaxime and Ciprofloxacin meanwhile 

2/10 (20%) E.coli isolates from Bian and Mansoura farm were 

sensitive to only three drugs, they were Cefotaxime , 

Ciprofloxacin and Doxycyclin for Bian farm meanwhile they 

were Cefotaxime , Ciprofloxacin and Gentamycin for 

Mansoura farm, 1/10 (10%) Salmonella isolate from Setawy 

farm was sensitive to only four drugs, they were Cefotaxime , 

Cipro-floxacin, Erythromycin and Streptomycin, 

2/10 (20%) E.coli isolates from Shahen and Asher 3 

farm were sensitive to only five drugs, they were Cefotaxime, 

71

Ciprofloxacin, Doxycyclin , Rifampicin and Streptomycin for 

Shahen farm meanwhile they were Ciprofloxacin, Cefotaxime, 

Gentamycin, Rifampicin, Streptomycin for Asher3 farm. 

2/10 (20%) E.coli isolates from Atef Abd El-Aziz and 

Asher 2 farm were sensitive to only six drugs, they were 

Cefotaxime ,Ciprofloxacin, Doxycyclin, Erythromycin, 

Rifampicin, Strepto-mycin for Atef Abd El-Aziz farm meanwhile 

they were Cefotaxime, Ciprofloxacin, Doxycyclin, Rifampicin, 

Streptomycin and Amp-icillin for Asher2 farm. 

1/10 (10%) E.coli isolate from Bela Bela farm was 

sensitive to seven drugs they were Cefotaxime, Ciprofloxacin , 

Gentamycin, Spectam, Colistin, Rifampicin and Doxycyclin. 

No isolates was sensitive to only eight or nine or ten drugs. 

72

Table(12 )Sensitivity Pattern of isolates from broiler. 

Sensitivity of microorganisms 

to only one or more drugs 

Sensitive to all drugs 

Sensitive to only one drug 

Sensitive to only two drugs 

Sensitive to only three drugs 

Sensitive to only four drugs 

Sensitive to only five drugs 

Sensitive to only six drugs 

Sensitive to only seven drugs 

Sensitive to only eight drugs 

Sensitive to only nine drugs 

Sensitive to only ten drugs 

Microorganisms 

Salmonella 

Nil 

E.coli 

E.coli 

Salmonella 

E.coli 

E.coli 

E.coli 

Nil 

Nil 

Nil 

Percentag 

e 

1/10 

(10%) 

0.0 

1/10 

(10%) 

2/10 

(20%) 

1/10 

(10%) 

2/10 

(20%) 

2/10 

(20%) 

1/10 

(10%) 

0.0 

4.13-Sensitivity percentages of each antimicrobial 

agent 

Results of sensitivity percentage of each antimicrobial 

agent against broiler isolates (table 13) showed that the most 

73 

0.0 

0.0

effective chemotherapeutic agent was cefotaxim with 

percentage 100% ,meanwhile 

(Sulphamethozole/Trimethoprime) was the lowest effective 

chemotherapeutic agent with percentage 10%, Ciprofloxacin 

recorded 90% sensitivity percentage so it comes after 

Cefotaxime in usage; Rifampicin recorded 70% sensitivity 

percentage, (Gentamycin, Spectam, Colistin, Erythromycin, 

Streptomycin, Ampicillin and Doxycyclin were recorded as 

intermediate effective chemotherapeutic agents. 


agent against one day old chicks isolates(fig.7) showed that 

the most effective chemotherapeutic agents were Cefotaxim 

and Ciprofloxacin that recorded 100% sensitivity meanwhile 

(Sulphamethazole/ Trimetho-prime) recorded the lowest 

sensitivity percentage that was 21.42%. Colistin and 

Gentamycin recorded sensitivity percentage 92.85% and 

85.71% respectively. 

Doxycyclin, Streptomycin, Amoxycyclin, Riphampicin and 

Erythro-mycin recorded sensitivity percentages 71.42%, 

71.42%, 28.57%, 64.28% and 57.14% respectively so they 

had intermediate effect. 


agents against isolates from layers revealed that cefotaxime, 

ciprofloxacin, rifa-mpicin, ampicillin, (Sulphamethazole+ 

Trimethoprime), genta-mycin, spectam, colistin, streptomycin, 

74

Doxycyclin, Erythromycin, were effective drugs respectively 

with percentages 100%, 100%, 63.63%, 63.63%, 63.63%, 

54.54%, 54.54%, 54.54%, 54.54% and 18.18%. 


agents against isolates from broiler revealed that cefotaxime, 

cipro-floxacin, rifampicin, doxycyclins, streptomycin, , 

gentamycin, erythromycin, spectam, ampicillin ,triple sulpha, 

amoxicillin, were effective with percentages 100%, 90%, 

70.0%, 60.0%, 60.0%, 40.0%, 30.0%, 20.0%, 20.0% and 

20.0% respectively 

Table (13) Sensitivity percentages of each antimicrobial 

against isolates from broiler, One day old chicks and layers. 

Antimicrobial 

agents 

Sensitivity percentages of each 

antimicrobial 

One day old 

chicks 

Broiler 

Cefotaxime 14/14(100%) 

Ciprofloxacin 14/14100% 

10/10(10%) 

9/10(90% 

) 

Gentamycin 12/14(85.71%) 4/10(40% 

) 

Sulphamethaz 3/14(21.42%) 1/10(10% 

ole 

/Trimethoprime 

) 

Spectam ND 2/10(20% 

) 

Colistin 13/14(92.85%) 2/10(20% 

) 

Erythromycin 8/14(57.14%) 3/10(30% 

) 

75 

Layer 

11/11(100%) 

11/11(100%) 

6/11(54.54%) 

7/11(63.63)% 

5/11(54.5%) 

2/11(18.18%) 

9/11(81.81%)

% 

120% 

100% 

80% 

60% 

40% 

20% 

0% 

Rifampicin 9/14(64.28%) 7/10(70% 

) 

7/11(63.63%) 

Streptomycin 10/14(71.42% 6/10(60% 6/11(54.54%) 

) 

) 

Ampicillin ND 2/10(20% 

) 

7/11(63.63%) 

Doxycyclin 10/14(71.42% 6/10(60% 6/11(54.54%) 

) 

) 

Amoxycillin 4/14(28.57%) ND ND. 

ND=not done 

Sensitivity percent 

Cefotaxime 

Ciprofloxacin 

Gentamycin 

(Sulphamethazole+ 

Trimethoprime) 

Spectam 

Colistin 

Erythromycin 

Riphampicin 

Streptomycin 

Ampicillin 

Doxycyclin 

Amoxycillin 

Agent 

76 

Broiler 

1 Day Chick 

Layer

Fig.(8) Sensitivity percentages of each antimicrobial agent 

against broiler, One day old chick and layer isolates. 

4.14-Results of antibiogram on isolates from human 

urine 

Concerning child urine amikacin and gentamycin were the 

most effective antibiotics out of all examined antibiotics(table 

14) ,mean-while amikacin, ciprofloxacin, gentamycin and 

nitrofurantoin respectively were the most effective against 

isolates obtained from male and femal urine isolates(Fig.8,9). 

Table (14) Antibogram E.coli isolates from human urine. 

Source Sample 

No. 

CTX30 CIP Amox GN F SXT25 ِAK RD 

1 R N R S N S S R 

Child 

Urine 2 S N S S N R S R 

Male 

Urine 

3 

1 

2 

3 

R N 

R R N R S R 

R S R S S R S R 

R R R S 

R R S 

R R R S R S S R 

77 

R

Femal 

urine 

4 

5 

1 

2 

3 

4 

5 

R S R R R R S R 

R S R S S R R R 

R R R S 

78 

S R S 

R R R S R R S R 

R S R S R R S R 

R R R S S R S R 

R S R R R R R R 

R = Resistant S = Sensitive N=not done CTX30 = Cefotaxime 

CIP= =Ciprofloxacin GN = Gentamycin SXT25 = Sulfphamethaxazole 

, Trimethoprime RD = Rifampicin. AK=amikacin 

Amox=amoxicillin F=nitrofurantoin 

R

Discussion 

Concerning bacteriological identification ,the obtained results revealed 

that 205 E.coli isolates were obtained out of 245 apparently healthy 

birds, this result agreed with Calnek.,(1997),who considered the E.coli 

as one of the serious problems responsible for economic losses to 

poultry industry. Meanwhile salmonella isolates represented 13.55%, 

comparable results were recorded by Craven et al.,(2000) who stated 

that 24% of broiler specimens were positive for salmonella species. 

Results of disk diffusion sensitivity tests against the bacterial 

isolates obtained from broiler revealed that all isolates obtained from 

all farms table were sensitive to cefotaxime, as concerning results 

which showed that the most effective antimicrobial agent against all 

tested E.coli broiler isolates was cefotaxime (100%), meanwhile lower 

percentage of cefotaxime sensitivity (62%) was recorded by Sylvester 

et al.,(2006). 

4/10(40%) E.coli from broiler were sensitive to gentamycin 

(table 9). Comparable results was obtained by Mishra et al.,(2002), 

increasing trend of resistance to gentamycin was also recoded by 

Lambie et al.,(2000). 

Except one E.coli broiler isolate (table 9), all broiler isolates 

were resistant to (sulphamethoxazole /trimethoprime), these results 

agreed with that obtained by Sepethri and Zadeh.,(2006) who found 

77

that the incidence of (sulphamethazole/trimethoprime) resistance 

among E.coli isolates was (80.7%), similar results obtained by Torkey 

(1995) who found that the resistance to (sulphamethazole 

/trimethoprime) was 88.5%. 

Spectam was more effective than (sulphamethazole/trimetho- 

prime) as antimicrobial agent ( table 9) where 2/10(20%) E.coli broiler 

isolates were sensitive to spectam, this result agreed with the results 

obtained by Eltahawy (1997) who noted that all Gram-negative bacilli 

isolates were resistant to at least two of the four major antibiotic groups 

(aminoglocoside, fluoroquinolon, third generation cephalosporin and 

carbapenems). 

Two out of ten (20%) E.coli broiler isolates were sensitive to 

colistin. Comparable results were found by Osman (1992) who 

reported 19.5% as the activity percentage of colistin against E.coli 

isolates. 

Six out of ten (60%) E.coli broiler isolates were sensitive to 

doxycyclin, meanwhile higher results were obtained by Saleh (2007) 

and Shome et al.,(2005), they reported that E.coli isolates were 

resistant to doxycyclin with percentage 96.6% and 100% respectively. 

On the other hand, Mishra (1995) found that all E.coli isolates 

collected from 2 commercial units and from free range chickens of 3 

villages were sensitive to chlortetracycline and doxycyclin. Vakani et 

78

al., (1997) reported that 10% of E.coli isolates were susceptible to 

doxycyclin. 

Seven of ten (70%) E.coli broiler isolates were resistant to 

erythromycin (table 9). Meanwhile Abd-El-Mawla (1998) recorded 

resistance percentage 100% against erythromycin and pencilin G. 

Seven of ten (70%) E.coli isolates were sensitive to 

rifampicin. Comparable results were obtained by Prescoet and 

Yielding (1990). 

Six out of ten (60%) E.coli isolates were sensitive to 

streptomycin. Similar results were obtained by Abd-El-Mawla (1998), 

Sylvester et al., (2006) and Khoshkhoo and Peighambari (2005) 

who recorded variable values of 60.9%, 71% and 26.7% respect- 

ively. 

Two out of ten(20%) E.coli broiler isolates were sensitive to 

ampicillin. Comparable sensitivity was obtained by David et al., 

(1996) who reported sensitivity percentage 16.9% to ampicillin, also to 

that obtained by Vakani et al., (1997) who found that less than 10% of 

E.coli isolates were sensitive to ampicillin. A study achieved on E.coli 

of poultry by Mishra et al.,(2002) showed the resistance of isolates to 

ampicillin 82%. 

79

Our study revealed that, all Salmonella broiler isolates were 

sensitive to cefotaxime. This result agreed with Juncosa morros et al., 

(2005) who reported that cefotaxime sensitivity percentage for 

Salmonella isolates was (99.9%). 

Also all Salmonella broiler isolates 100% were sensitive to 

ciprofloxacin. This result also stimulated by Babireke-Iriso et al., 

(2006) who found that sensitivity to ciprofloxacin was 97%. 

1/2(50%) Salmonella broiler isolate was sensitive to 

gentamycin. Meanwhile higher results were obtained by Lee et 

al.,(2003) who found that gentamycin sensitivity was 43%. Our lower 

results might be due to few obtained strains of salmonellae. 

1/2(50%) Salmonella broiler isolate was sensitive to(sulpha- 

methazole /trimethoprime). This result agreed with Larkin et 

al.,(2004) who found that all salmonella isolates were sensitive to 

(cephalothin, chloramphenicol, ciprofloxacin, gentamycin, sulpha- 

methazole /trimethoprime and tetracycline) by varible percentages. 

1/2(50%) Salmonella broiler isolate was sensitive to spectam, 

this result agreed with Eltahawy(1997) who noted that all Gram- 

negative bacilli were resistant to at least two of four major antibiotics. 

80

1/2(50%) Salmonella broiler isolates was sensitive to colistin, 

meanwhile Larkin et al.,(2004) who recorded activity percentage 

64.4% of colistin. Our lower results might be due to few obtained 

strains of salmonellae. 

1/2(50%) Salmonella broiler isolates was sensitive to 

doxycyclin. This result agreed Saleh (2007), who record 96.6% 

resistance percentage to doxycyclin. Kumar et al.,(2003) who reported 

resistance percentage 58.8% for doxycyclin, on the other hand, and all 

isolates were resistant to doxycyclin as reported by Hegazy (1991). 

Salmonella broiler isolates were sensitive to erythromycin. This 

result agreed with Kondrachi (1976) who mentioned that most strains 

were sensitive to erythromycin, neomycin, Similar result was obtained 

by Osman (1992) who reported that erythromycin sensitivity 

percentage was 73.8% ,meanwhile disagreed with Abbass (1993) who 

reported that erythromycin is not effective against E.coli. 

1/2(50%) Salmonella broiler isolates was sensitive to rifampicin 

this result agreed with that of Molla et al.,(1999) who found that only 

4 antimicrobials (gentamycin, kanamycin, rifampicin and 

sulphamethazole/ trimethoprime) were effective against Salmonella 

isolates. 

Salmonella broiler isolates were sensitive to streptomycin, 

meanwhile Kumar et al.,(2003) reported resistance percentage 

81

45.10%. Resistance percentage 11.9% was obtained by Kikuvi et 

al.,(2006). 

1/2(50%) Salmonella broiler isolates was sensitive to ampicillin. 

It was also found that the most common resistance was reported against 

ampicillin as reported by Kikuvi et al., (2006) and 77% resistance 

percentage to ampicillin as was recorded by Yang et al.,(2004). 

Antibiogram of isolates from one day old chicks revealed that:- 

14 E.coli from one day old chicks (100%) were sensitive to 

cefotaxime. Comparable results were stimulated by Saha et al.,(2003) 

who recorded cefotaxime sensitivity 79.17%, meanwhile lower 

resistance 62% was reported by Sylvester et al.,(2006). 

All 14 E.coli from one day old chicks were sensitive to 

ciprofloxacin. Meanwhile higher Ciprofloxacin resistance percentage 

50.98% was obtained by Kumer et al.,(2003). Vakani et al.,(1997) 

found that 46.27% of E.coli isolates from chickens were resistant to 

ciprofloxacin. 44% of avian E.coli isolates were found resistant to 

ciprofloxacin as reported by Khoshkhoo and Peighambari(2005). 

Four out of 14(28.57%) E.coli from one day old chicks were 

sensitive to amoxycillin. This result agreed with the result mentioned 

by Lambie et al.,(2000) who isolated E.coli from diseased broilers, the 

isolates showed increasing resistance to amoxycillin. 

82

12/14(85.71%) E.coli isolates from one day old chicks were 

sensitive to gentamycin, meaning that gentamycin resistance result was 

14.29%. Comparable results was stimulated by Sylvester et al.,(2006), 

Kumar et al.,(2003) who recorded gentamycin resistance percentage 

20%, 22.5% respectively, Resistance percentages 34.5% ,34% and 

33.4% were obtained by Saleh (2007),Archana et al.,(2002) and 

Osman (1992) respectively. 

Three out of 14 (21.42%) E.coli isolates from one day old chicks 

were sensitive to sulphamethazole/ trimethoprime means that 

resistance percentage was 78.58%, later result agreed with that 

mentioned by Torkey et al.,(1995), Abd-El-Mawla (1998) and 

Gholamreza and Abbass (2006) who found that the incidence of 

sulphamethazole/trimethoprime was 88.5%, 80.4% and 80.7% 

respectively, it disagreed with the percentage of sulphamethazole/ 

trimethoprime resistant avian E.coli isolates 67%, 92% and 72.6% as 

were obtained by Blanco et al.,(1997), Saleem et al.,(1999) and 

Khoshkhoo and Peighambari (2005) respectively. 

Therteen out of 14(92.85%) E.coli isolates from one day old 

chicks isolates were sensitive to colistin. This result agreed with that 

mentioned by Prescott and Yieldding (1990) who reported that 

fluoroqunimolones had potent antimicrobial activity against 

Enterobacteriaceae. 

83

Ten out of 14(71.42%) E.coli from one day old chicks isolates 

were sensitive to doxycyclin. Tsai and Fang found that over 90% of 

E.coli isolates were resistant to oxytetracycline, chlortetracycline, 

tetracycline and doxycyclin. Shome and his colleagues found that all 

E.coli isolates were also resistant, on the other hand, it was found all 

E.coli isolates collected from 2 commercial unites and from free range 

chickens from 3 villages were sensitive to chlortetracycline and 

doxycyclin, Mishra(1995). 

Eight out of 14(57.14%) E.coli isolates from one day old chicks 

were sensitive to erythromycin, Activity percentage 73.8% of 

erythromycin was mentioned by Osman (1992), meanwhile resistance 

percentage 100% against erythromycin and pencillin G was recorded 

by Abd-El-Mawla (1998). 

Nine out of 14(64.28%) E.coli isolates from one day old chicks 

were sensitive to rifampicin. This result was similarly stimulated by 

Prescott and Yieldding (1990). 

Ten out of 14(71.42%) E.coli isolates from one day old chicks 

were sensitive to streptomycin. Meaning that streptomycin resistance 

percentage was 28.58%. This result agreed with that obtained by Abd- 

El-Mawla (1998) who reported resistance percentage 39.1% for 

84

streptomycin, meanwhile higher percentage 81% was obtained by 

Adesiyum and Kaminjoli (1982). 

Our study revealed that all Salmonella isolated obtained from 

one day old chicks were sensitive to cefotaxime, comparable results 

were recorded by Jamal et al.,(1998) and Juncosa Morros et 

al.,(2005)who reported that cefotaxime sensitivity for Salmonella was 

99.7% and 99.9% respectively. 

All Salmonella isolates from one day old chicks were sensitive 

to ciprofloxacin, this result agreed with that obtained by Larkin et 

al.,(2004) who found that all Salmonella species isolates were sensitive 

to ciprofloxacin. 

Antibiogram of isolates from layers revealed that :- 

All E.coli isolates from layer were sensitive to cefotaxime. 

This results agreed with Liu et al.,(2007) who reported that E.coli 

isolates were more susceptible to cefotaxime than to ceftiofur. 

All E.coli isolates from layer were sensitive to ciprofloxacin. 

Ciprofloxacin resistance percentage 50.98% were obtained by Kumar 

et al.,(2003). Vakani et al.,(1997), found that 46.27% of Escherichia 

coli isolates from chicken were resistant to ciprofloxacin. 44% and 34% 

resistance percentages of E.coli isolates were recorded by Khoshkoo 

and Peighambari (2005) and Sylvester et al.,(2006) respectively. 

85

Seven out of eleven (63.63%) E.coli isolates from layers were 

sensitive to ampicillin. Comparable results were obtained by Abd-El- 

Mawla (1998) who recorded moderate sensitivity percentage 50% for 

ampicillin against E.coli isolates. Higher percentages 89.6% and 96.7% 

were obtained by Gundogan et al.,(2006) and Khoshkhoo and 

Peighambari (2005) respectively, meanwhile other studies on E.coli of 

poultry showed that the resistance of isolates to ampicillin were 82% 

and 74.24% as recorded by Mishra et al.,(2002) and Hui and Das 

(2000) respectively. 

Six out of eleven(54.54%) E.coli isolates from layers were 

sensitive to gentamycin. This result agreed with the result obtained by 

El-Ghamdi et al.,(1999) who compared antibiotic resistant E.coli 

isolates to ampicillin, chloramphenicol, gentamycin, spectinomycin, 

tetracyclin , trimethoprime spectinomycin, tetracyclin and sulpha- 

methazole and found it ranged from 57% to 99.1%. 


sensitive to sulphamethazole/ trimethoprime. This result is nearest to 

the result of El-Sayed et al.,(2001) who stated that sulphamethazole 

/trimethoprime resistance percentage was 73.3% and disagreed with 

Blanco et al.,(1997) who stated that resistance percentage was 67%. 

86

Two out of eleven(18.18%) E.coli isolates from layers were 

sensitive to colistin, on the other hand, Abd- El-Motelib and El- 

Zanati (1993). Giurov (1985) recorded the susceptibility percentage 

96.06% for 223 strains of E.coli using disk diffusion method. 


sensitive to doxycyclin, this result agreed with Van Donkersgoed et 

al.,(2003) who found that 68% E.coli isolates were sensitive to all 18 

antimicrobial tested agents. 

Nine out of eleven(81.81%) E.coli isolates from layer were 

sensitive to erythromycin. 73.8% activity percentage of erythromycin 

obtained by Osman (1992). Resistance percentage 100% of 

erythromycin was obtained by Abd-El-Mawla (1998). 


sensitive to rifampicin, this result agreed with Prescott and Yieldding 

(1990) who reported that fluoroquniolon had a potent antimicrobial 

activity against Enterobacteriaceae . 


sensitive to streptomycin, this result agreed with Giurov(1985) who 

obtained sensitivity percentage 44.84% to streptomycin. Abd-El- 

Mawla (1998) reported sensitivity percentage 39.1% to streptomycin, 

meanwhile Osman (1992) obtained sensitivity percentage 80.9%. 

87

Five out of eleven(54.5%) E.coli isolates from layers isolates 

were sensitive to spectam. this result agreed with that obtained by 

Eltahawy (1997) who noted that all Gram-negative bacilli isolates 

were resistant to at least two of the four major antibiotics group 

(aminoglocoside, fluoroquinolon, third generation cephalosporin and 

carbapenems). 

Our study revealed that all Salmonella isolates were sensitive to 

cefotaxime. This result agreed with Jamal et al.,( 1998) and Juncosa 

Morros et al., (2005) who reported that cefotaxime sensitivity for 

Salmonella isolates were 99.7% and 99.9% respectively. 

All Salmonella isolates from layer were sensitive to 

ciprofloxacin. This result agreed with Jacobs-Reitsma et al.,(1994), 

Poppa et al.,(1995) and Juncosa Morros et al., (2005) who found 

that all Salmonella isolates were sensitive to ciprofloxacin also, this 

result agreed with Babirake-Iriso et al.,(2006) who found that 

sensitivity to ciprofloxacin was 97%, variable values 91.5% and 

93.33%) were obtained by David et al.,(1996) and Hui and Das 

(2001) respectively. 

1 out of 2(50%) Salmonella isolates from layer were sensitive to 

ampicillin, this result agreed with that obtained by Giurov (1985) who 

recorded that ampicillin sensitivity percentage was 51.12% and Dinch 

88

and Nguyen (1995) who reported moderate sensitivity to ampicillin 

also, this result agreed with Abd-El-Mawla (1998) who recorded 

sensitivity percentage 50% for ampicillin. 


gentamycin this result agreed with that obtained by El-Ghamdi et 

al.,(1999) who recorded from resistance percentage ranged from 57% 

to 99.1% for gentamycin also, Lambie et al., (2000) reported that the 

isolates showing increasing trend of resistance to amoxicillin, 

paramecia, gentamycin, nitrofurance, norfloxacin and 

sulfamethazole/trimethoprime. 

No Salmonella isolates were sensitive to sulfamethazole/ 

trimethoprime. this means that sulfamethazole/trimethoprime resistance 

percentage was 100%. This result agreed with that obtained by Poope 

et al.,(1995) who recorded sensitivity percentage 1.7% for 

sulfamethazole /trimethoprime and agreed with Lambie et al., (2000) 

who reported resistance for sulpha. And disagreed with resistance 

percentage 73.3% which obtained by El-Sayed (2001). 

No Salmonella isolates were sensitive to colistin this means 

resistance percentage 100%, this result agreed with Saha et al (1992) 

who found that Salmonella typhimuriun isolates showed resistance 

against commonly used antimicrobial agents. 

89


to doxycyclin. Resistance percentage 96.6% to doxycyclin was 

recorded by Saleh (2007) on the other hand, all isolates were resistant 

to doxycyclin, as mentioned by Hegazy (1991). 

No Salmonella isolates were sensitive to erythromycin this 

means resistance percentage 100%, this result agreed with Abbass 

(1993) who reported that colistin, sulphate, erythromycim, lincomycin 

and pencillin were not effective. 

Two Salmonella isolates from layer were sensitive to 

rifampicin. This result agreed with Molla et al.,(1999) who found that 

only 4 antimicrobials ( gentamycin, kanamycin, rifampicin and 

sulfamethazole/ trimethoprime ) were effective against all Salmonella 

isolates. 

No Salmonella isolates were sensitive to streptomycin this 

means that resistance percentage 100%, this result agreed with Jain 

and Chen (2006) who found that 98.9% of Salmonella isolates were 

resistant to streptomycin. Saleh (2007) found that resistance percentage 

79.1% to streptomycin. 

All Salmonella isolates from layer were resistant to spectam. 

This result agreed with Eltahawy (1997) who noted that all Gram- 

negative bacilli isolates were resistant to at least two of the four major 

antibiotics group (aminoglocoside, fluoroquinolon, third generation 

cephalosporin and carbapenems). 

90

Summary 

Bacterial diseases have long been recognized as a major risk factor in 

poultry health management. Every poultry farmer is aware of the risks 

of bacterial infections and their subsequent effect on mortalities, 

productivity and profitability. The present work could be summarized 

in the following:- 

91

1- Enterobacteria were isolated from liver and spleen of chickens 

(broiler, one day old chicks and layers) with prevalence rate 83.05%. 

69.49% and 13.55% isolates were lactose fermenter and non-lactose 

fermenter respectively. 

2-Incidence of lactose fermenter among enterobacteria were 61.6% 

and 81.1% in broiler and in both layers and one day old chicks 

respectively, meanwhile Incidence of non lactose fermenter among 

enterobacteria were 24% and 5.88% in both layers and one day old 

chicks respectively, 

3-lactose fermenter isolates from human urine (female, male and 

children) represented 29.8%, 18/30 81.8% and 60% of total samples 

respectively, meanwhile non lactose fermenter isolates represented 

5.5%, 13.33% and 0.0(0.0%) of total samples respectively, 

4-Lactose fermenter isolates from broiler were E.coli and Citrobacter 

diversus among tested isolates and represented 68/107 (63.55%) and 

9/107 (8.41%) respectively. Meanwhile non Lactose fermenter isolates 

were salmonella, pseudomonas and proteus and represented 24/107 

(22.42%), 4/107(3.7%), 2/107(1.86%) respectively 

5-Lactose fermenter isolates from one day old chicks and layers (table 

4) were E.coli and Citrobacter diversus among tested isolates. They 

92

epresented 105/138(76.08%) and 33/138(23.91%) respect-ively. 

Meanwhile non Lactose fermenter isolates were salmonella, 

pseudomonas and proteus and represented 7/138 (5.07%), 2/138 (1.44%) 

and1/138 (0.27%) respectively. 

6-Lactose fermenter isolates from males urine subjects (table 4) were 

E.coli and Citrobacter diversus among tested isolates and represented 

16/18 (88.88%) and 2/18 (11.11%) respectively. Meanwhile non Lactose 

fermenter isolates were salmonella, pseudomonas and proteus and 

represented 3/4 (75%),1/4(25%) and 0.0 respectively. 

7-Lactose fermenter isolates from childe urine subjects(table 4) were 

E.coli and Citrobacter diversus among tested isolates and represented 

3/3(100%) , 0.0 and 0.0 respectively. Meanwhile non Lactose fermenter 

isolates were salmonella, pseudomonas and proteus and represented 0.0, 

0.0 and 0.0 respectively 

8-Results of antibiogram on biotypes from one day old chicks showed 

that all the isolates were sensitive to cefotaxime and ciprofloxacin also 

all farms isolates were sensitive to colistin except one isolate from 

only one farm. 

9-Results of sensitivity pattern of the isolates from one day old chicks 

revealed that most isolates were sensitive to three or more drugs. 

Fortunately there were no isolates sensitive to only one or two drugs. 

93

10-Results of sensitivity percentage of each antimicrobial agents 

against isolates from one day old chicks revealed that cefotaxime, 

ciprofloxacin, colistin, gentamycin, streptomycin, doxycyclins, 

rifampicin, erythromycin triple sulpha, amoxicillin, were effective with 

percentages 100%, 100%, 92.85%, 71.42%, 71.42%, 64.28%, 

64.28%, 57.14% and 28.57% respectively. 

11-Results of antibiogram on layers isolates revealed that the isolates 

from all farms were sensitive to cefotaxime and ciprofloxacin 

meanwhile the isolates from all farms were resistant to colistin except 

only two farms. 

12- Sensitivity Pattern of layers isolates revealed that there were no 

isolates sensitive to all drugs also no isolates were sensitive to only one 

or only two or only three drugs, meanwhile most isolates were sensitive 

to only four or more drugs indicating that the isolates from layers were 

less resistant than that from one day old chicks. 


against isolates from layers revealed that cefotaxime, ciprofloxacin, 

rifampicin, ampicillin, (Sulphamethazole+ Trimethoprime), 

gentamycin, spectam, colistin, streptomycin, Doxycyclin, 

Erythromycin, were effective drugs respectively with percentages 

94

100%, 100%, 63.63%, 63.63%, 63.63%, 54.54%, 54.54%, 54.54%, 

54.54% and 18.18%. 

14-Results of antibiogram on isolates from broiler revealed that all 

isolates were sensitive to cefotaxime meanwhile there was varying 

resistance to (Sulphamethazole, trimethoprime), Colistin and Spectam. 


against isolates from broiler revealed that cefotaxime, ciprofloxacin, 

rifampicin, doxycyclins, streptomycin, , gentamycin, erythromycin, 

spectam, ampicillin ,triple sulpha, amoxicillin, were effective with 

percentages 100%, 90%, 70.0%, 60.0%, 60.0%, 40.0%, 30.0%, 20.0%, 

20.0% and 20.0% respectively. 

16-Results of sensitivity pattern of the isolates from broiler revealed 

that except one isolate, most isolates were sensitive to only two or 

more drugs indicating that the isolates from broiler were more resistant 

than that from layers but they less resistant than that from one day old 

chicks. 

17-Results of antibiogram on biotypes from human urine revealed the 

following:-Concerning children urine, amikacin and gentamycin were 

the most effective antibiotics out of all examined antibiotics, 

meanwhile amikacin, ciprofloxacin and gentamycin were the most 

effective against isolates obtained from male and female urine isolates. 

95

Conclussion 

1- Enterobacteria were isolated from broiler, one day old chicks and 

layers with prevalence rate 83.05%. lactose fermenter were frequently 

isolated from all examined specimens of chickens more than non-lactose 

fermenter.It was similarly recorded in human urine specimens. 

2-Lactose fermenter isolates from broiler were E.coli and Citrobacter 

diversus. Meanwhile non Lactose fermenter isolates were salmonella, 

pseudomonas and proteus. 

96

3-Results of antibiogram on biotypes from one day old chicks, broiler and 

layers showed that all the isolates were sensitive to cefotaxime and 

ciprofloxacin, also all one day old chicks isolates were sensitive to colistin 

except one isolate from only one farm, meanwhile the isolates from all layer 

farms were resistant to colistin except only two farms and there was 

varying resistance to (Sulphamethazole, trimethoprime), Colistin and 

Spectam in broiler isolates. 

4-Sensitivity Pattern of layers isolates revealed that there were no isolates 

sensitive to all drugs also no isolates were sensitive to only one or only two 

or only three drugs, meanwhile most isolates were sensitive to only four or 

more drugs indicating that the isolates from layers were less resistant than 

that from one day old chicks. 

5-Results of sensitivity percentage of each antimicrobial agents against 

isolates from one day old chicks revealed that the cefotaxime, ciprofloxacin, 

and colistin are first three effective drugs should used in cases of 

emergencies, infection control or for treatment of one day old chicks. 

6-Results of sensitivity percentage of each antimicrobial agents against 

isolates from layers and broiler revealed that the cefotaxime, ciprofloxacin 

and rifampicin, are first three effective drugs should used in cases 

emergencies, infection control or for treatment of layers and broiler. 

97

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118 --

ﻰﺑﺮﻌﻟا ﺺﺨﻠﻤﻟا 

ﺢﺒﺻاو ﻦﺟاوﺪﻟا ﺔﺤﺻ ﻰﻠﻋ ةرﻮﻄﺧ تاذ ﺔﻳﺮﻴﺘﻜﺒﻟا ضاﺮﻣﻻا ﺮﺒﺘﻌﺗ 

ﻰﻠﻋ ﺎﻬﺗﺎﻌﺒﺗو ﺔﻳﺮﻴﺘﻜﺒﻟا تﺎﺑﺎﺻﻻا ﺪﻳاﺰﺗ ﻦﻣ فﻮﺨﺗ ﻰﻓ ﻦﺟاوﺪﻟا ﻰﺠﺘﻨﻣ 

ﺪﻟاﻮﺗ ﻰﻟا ﺔﻳﺮﻴﺘﻜﺒﻟا ﺔﺑﺎﺻﻻا راﺮﻜﺗ ﻊﺟﺮﻳو . قﻮﻔﻨﻟا ةدﺎﻳزو ﺔﻴﺟﺎﺘﻧﻻا 

ﻞﻤﻌﻟا اﺬه ﻰﻓو ﺔﻳﻮﻴﺤﻟا تادﺎﻀﻤﻠﻟ ﺔﻣوﺎﻘﻤﻟا ةدﺪﻌﺘﻣ ﺎﻳﺮﺘﻜﺒﻟا ﻦﻣ عاﻮﻧا 

. ﺎهﺪﺟاﻮﺗ ىﺪﻣو 

عاﻮﻧﻻا ةﺬه ﺪﻳﺪﺤﺗ ﻢﺗ 

جﺎﺟد ىراﺪﺑ ﺐﻠﻗو لﺎﺤﻃو ﺪﺒآ ﻦﻣ ﺔﻳﻮﻌﻣ فﻮﺠﻟا ﺎﻳﺮﻴﺘﻜﺒﻟا لﺰﻋ ﻢﺗ -1 

عاﻮﻧﻻا ﺖﻧﺎآو . ﺪﺣاو مﻮﻳ ﺮﻤﻋ ﺖﻴآﺎﺘﻜﻟاو ضﺎﻴﺒﻟا جﺎﺟﺪﻟاو ﻦﻴﻤﺴﺘﻟا 

% 83.6 ﺔﺒﺴﻨﺑ ﺎﻬﻟﺰﻋ ﻢﺗ ﺚﻴﺣ زﻮﺘآﻻا ﺮﻤﺨﺗ ﻰﺘﻟا ﻰه اﺪﺟاﻮﺗ ﺮﺜآﻻا 

ىﻻﻮآ ﺎﻴﺷﺮﺸﻳﻻا ﺖﻧﺎآ .% 16.32 ﺔﺒﺴﻨﺑ 

ﺖﻧﺎآ ةﺮﻤﺨﺗﻻ ﻰﺘﻟا ﺎﻤﻨﻴﺑ 

لﺰﻋ ﻢﺗ ﺎﻤﻨﻴﺑ عراﺰﻤﻟا ﺔﻴﺒﻟﺎﻏ ﻦﻣ ﺎﻬﻟﺰﻋ ﻢﺗ ﺚﻴﺣ اﺪﺟاﻮﺗ عاﻮﻧﻻا ﺮﺜآا 

ىﻻﻮآ ﺎﻴﺷﺮﺸﻳﻻا ﻦﻣ ﻞآ لﺰﻋ ﻢﺗ ﺪﻗو . عراﺰﻤﻟا ﺾﻌﺑ ﻦﻣ ﻼﻴﻧﻮﻤﻟﺎﺴﻟا 

. ﺮﺑﺎﻨﻌﻟا ﺲﻔﻧ ﺾﻌﺑ ﻦﻣ ﻼﻴﻧﻮﻤﻟﺎﺴﻟا و 

ﺖﻠﺜﻣ ﺚﻴﺣ نﺎﺴﻧﻻا لﻮﺑ ﻦﻣ ﺔﻳﻮﻌﻣ فﻮﺠﻟا ﺎﻳﺮﻴﺘﻜﺒﻟا لﺰﻋ ﻢﺗ -2 

لﺎﺟﺮﻟﺎﺑ ﺔﺻﺎﺨﻟا تﺎﻨﻴﻌﻟا ﻰﻓ ﺎﻬﻨﻣ ﺮﺒآﻻا ءﺰﺠﻟا ىﻻﻮآ ﺎﻴﺷﺮﺸﻳﻻا 

. تاﺪﻴﺴﻟا ﻰﻓ ﺐﻠﻏﻻا ﻮه ﻼﻴﻧﻮﻤﻟﺎﺴﻟا عﻮﻧ نﺎآ ﺎﻤﻨﻴﺑ , لﺎﻔﻃﻻاو ءﺎﺴﻨﻟاو 

ﻦﻣ ﺔﻟوﺰﻌﻤﻟا ﺎﻳﺮﻴﺘﻜﺒﻠﻟ ﺔﻳﻮﻴﺤﻟا تادﺎﻀﻤﻠﻟ ﺔﻴﺳﺎﺴﺤﻟا تارﺎﺒﺘﺧا ﺞﺋﺎﺘﻧ-3 

ﻞﻜﻟ ﺔﻳﺮﻴﺘﻜﺒﻟا تاﺮﺘﻌﻟا ﻞآ ﺔﻴﺳﺎﺴﺣ تﺮﻬﻇا ﺪﺣاو مﻮﻳ ﺮﻤﻋ ﺖﻴآﺎﺘﻜﻟا 

ﺮﻴﺛﺎﺗ ﻦﻴﺘﺴﻟﻮﻜﻠﻟ نﺎآ ﺪﻘﻟو ﻦﻴﺳﺎﺴآﻮﻠﻓوﺮﺒﺴﻟاو ﻢﻴﺴآﺎﺗﻮﻔﻴﺴﻟا ﻰﺋاود ﻦﻣ 

. ﺪﺣاو ﺮﺒﻨﻋ ءﺎﻨﺜﺘﺘﺳﺎﺑ ﺮﺑﺎﻨﻌﻟا ﻞآ ﻰﻠﻋ ﻖﻠﻄﻣ 

مﻮﻳ ﺮﻤﻋ ﺖﻴآﺎﺘﻜﻟا ﻦﻣ ﺔﻟوﺰﻌﻤﻟا تﺎﺑوﺮﻜﻴﻤﻠﻟ ﺔﻴﺳﺎﺴﺤﻟا ﻂﻤﻧ نﺎآ -4 

ﻦﻣ ﺮﺜآا وا ﺔﺛﻼﺜﻟ ﺔﺳﺎﺴﺣ ﺖﻧﺎآ تﻻﺰﻌﻟا ﻢﻈﻌﻣ نا ﺎﺤﺿﻮﻣ ﺪﺣاو 

ﺔﺳارﺪﻠﻟ ﺔﻌﺿﺎﺨﻟا ﺔﻳﻮﻴﺤﻟا تادﺎﻀﻤﻟا

ﻦﻴﺘﺴﻟﻮآ , ﻦﻴﺳﺎﺴآﻮﻠﻓوﺮﺒﺳ , ﻢﻴﺴآﺎﺗﻮﻔﻴﺳ ﺔﻠﻣﺎﺷ ﺔﻳﻮﻴﺤﻟا تادﺎﻀﻤﻟا 

-5 

, ﻦﻴﺴﻴﻣوﺮﺜﻳرا , ﻦﻴﺴﻴﺒﻣﺎﻔﻳر , ﻦﻴﺴﻴﻣﻮﺘﺑﺮﺘﺳ , ﻦﻴﻠﻠﻴﺴﺒﻣا , ﻦﻴﺴﻴﻣﺎﺘﻨﺟ , 

, % 100 % 100 ﺐﺴﻨﺑ لﺎﻌﻓ ﺮﻴﺛﺎﺗ تﺮﻬﻇا ﺎﻔﻠﺴﻟاو ﻦﻴﻠﻠﻴﺴﻴﺴآﻮﻣا 

% 21.42, 

% 57.14 , % 64.28, 

% 64.28 , % 71.42, 

% 92.85 

. ﺪﺣاو مﻮﻳ ﺮﻤﻋ ﺖﻴآﺎﺘﻜﻟا 

ﻦﻣ ﺔﻟوﺰﻌﻤﻟا تﺎﺑوﺮﻜﻴﻤﻠﻟ ﺪﺿ 

ﻦﻣ ﺔﻟوﺰﻌﻤﻟا ﺎﻳﺮﻴﺘﻜﺒﻠﻟ ﺔﻳﻮﻴﺤﻟا تادﺎﻀﻤﻠﻟ ﺔﻴﺳﺎﺴﺤﻟا تارﺎﺒﺘﺧا ﺞﺋﺎﺘﻧ-6 

ﻰﺋاود ﻦﻣ ﻞﻜﻟ ﺔﻳﺮﻴﺘﻜﺒﻟا تاﺮﺘﻌﻟا ﻞآ ﺔﻴﺳﺎﺴﺣ تﺮﻬﻇا ضﺎﻴﺒﻟا جﺎﺟﺪﻟا 

ﻰﻓ ﻦﻴﺘﺴﻟﻮﻜﻠﻟ ﺔﻣوﺎﻘﻣ كﺎﻨه نﺎآ ﺪﻘﻟو ﻦﻴﺳﺎﺴآﻮﻠﻓوﺮﺒﺴﻟاو ﻢﻴﺴآﺎﺗﻮﻔﻴﺴﻟا 

. ﻦﻳﺮﺒﻨﻋ ءﺎﻨﺜﺘﺳﺎﺑ ﺮﺑﺎﻨﻌﻟا ﻞآ 

ضﺎﻴﺒﻟا جﺎﺟﺪﻟا ﻦﻣ ﺔﻟوﺰﻌﻤﻟا تﺎﺑوﺮﻜﻴﻤﻠﻟ ﺔﻴﺳﺎﺴﺤﻟا ﻂﻤﻧ نﺎآ -7 

ﻦﻣ ﻂﻘﻓ ﺔﺴﻤﺧ وا ﺔﻌﺑرﻻ ﺔﺳﺎﺴﺣ ﺖﻧﺎآ تﻻﺰﻌﻟا ﻢﻈﻌﻣ نا ﺎﺤﺿﻮﻣ 

ﺎﻬﺗﺮﻴﻈﻧ ﻦﻣ ﺔﻣوﺎﻘﻣ ﻞﻗا تﻻﺰﻌﻟا ةﺬه ﺖﻧﺎآ ﻚﻟﺬﺑو ﺔﻳﻮﻴﺤﻟا تادﺎﻀﻤﻟا 

. ﺪﺣاو مﻮﻳ ﺮﻤﻋ ﺖﻴآﺎﺘﻜﻟا ﻦﻣ ﺔﻟوﺰﻌﻤﻟا 

, ﻦﻴﺳﺎﺴآﻮﻠﻓوﺮﺒﺳ 

, ﻢﻴﺴآﺎﺗﻮﻔﻴﺳ ﺔﻠﻣﺎﺷ ﺔﻳﻮﻴﺤﻟا تادﺎﻀﻤﻟا -8 

, ﻦﻴﺴﻴﺒﻣﺎﻔﻳر , ﻦﻴﺴﻴﻣوﺮﺜﻳرا , ﻦﻴﺘﺴﻟﻮآ , مﺎﺘﻜﺒﺳ , ﺎﻔﻠﺴﻟا , ﻦﻴﺴﻴﻣﺎﺘﻨﺟ 

ﺐﺴﻨﺑ لﺎﻌﻓ ﺮﻴﺛﺎﺗ تﺮﻬﻇا ﻦﻴﻠﻜﻴﺴﻴﺴآودو , ﻦﻴﻠﻠﻴﺴﺒﻣا , ﻦﻴﺴﻴﻣﻮﺘﺑﺮﺘﺳ 

% 36.6, 

% 36.6, 

% 54.5, 

% 45.4, 

% 18.1, 

% 72.7, 

% 00 % 100 

ﺖﻴآﺎﺘﻜﻟا ﻦﻣ ﺔﻟوﺰﻌﻤﻟا تﺎﺑوﺮﻜﻴﻤﻟا ﺪﺿ % 72.7, 

% , 36.6% 

,45.6 

. ﺪﺣاو مﻮﻳ ﺮﻤﻋ 

ﻦﻣ ﺔﻟوﺰﻌﻤﻟا ﺎﻳﺮﻴﺘﻜﺒﻠﻟ ﺔﻳﻮﻴﺤﻟا تادﺎﻀﻤﻠﻟ ﺔﻴﺳﺎﺴﺤﻟا تارﺎﺒﺘﺧا ﺞﺋﺎﺘﻧ -9 

ءاوﺪﻟ ﺔﻳﺮﻴﺘﻜﺒﻟا تاﺮﺘﻌﻟا ﻞآ ﺔﻴﺳﺎﺴﺣ تﺮﻬﻇا ﻦﻴﻤﺴﺘﻟا ىراﺪﺑ 

ﺎﻔﻠﺴﻟاو ﻦﻴﺘﺴﻟﻮﻜﻠﻟ ﺔﻨﻳﺎﺒﺘﻣ ﺔﻣوﺎﻘﻣ كﺎﻨه نﺎآ ﺪﻘﻟو ﻢﻴﺴآﺎﺗﻮﻔﻴﺴﻟا 

. 

مﺎﺘﻜﺒﺴﻟاو

ﻦﻣ 

ﺔﻟوﺰﻌﻤﻟا ﺎﻳﺮﻴﺘﻜﺒﻠﻟ ﺔﻳﻮﻴﺤﻟا تادﺎﻀﻤﻠﻟ ﺔﻴﺳﺎﺴﺤﻟا تارﺎﺒﺘﺧا ﺞﺋﺎﺘﻧ -9 

ﻦﻴﺳﺎﻜﻴﻣﻻا ﻰﺋاود نﺎآ لﺎﻔﻃﻻا تﺎﻨﻴﻌﺑ ﻖﻠﻌﺘﻳ ﺎﻤﻴﻓ -: 

ﻰﺗﻻا تﺮﻬﻇا لﻮﺒﻟا 

نﺎآ ﺎﻤﻨﻴﺑ ﺺﺤﻔﻠﻟ ﺔﻌﺿﺎﺨﻟا ﺔﻳودﻻا ﻦﻤﺿ ﺔﻴﻟﺎﻌﻓ ﺮﺜآﻻا ﺎﻤه ﻦﻴﺴﻴﻣﺎﺘﻨﺠﻟاو 

ﺪﺿ ﺔﻴﻟﺎﻌﻓ ﺮﺜآﻻا ﺎﻤه ﻦﻴﺴﻴﻣﺎﺘﻨﺠﻟاو ﻦﻴﺳﺎﺴآﻮﻠﻓوﺮﺒﺴﻟاو ﻦﻴﺳﺎﻜﻴﻣﻻا 

. ثﺎﻧﻻاو رﻮآﺬﻟا تﺎﻨﻴﻋ ﻦﻣ ﺔﻟوﺰﻌﻤﻟا تﺎﺑوﺮﻜﻴﻤﻟا 

VITA 

- The auther was born on 30 October 1984 in Zagazig, Sharkia.

- She received her primary education in kawmia primary school, 

Zagazig from which graduated in 1994. 

- She received her preparatory education in EL-Sadat prep. 

School for girls, Zagazig from which graduated in 1997. 

- She received her secondary education in EL-Sadat secondary 

School for girls, Zagazig from which graduated in 2000. 

- She graduated from Faculty of Science, Zagazig University in 

2005. 

- She occupied in research institute of animal health in 2006. 

- She applied for the degree of Master in bacteriology in faculty 

of Vet. Med., Zagazig in 9/9/2005.

E. Coli

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