Avian Clostridial

Infections and Gut

IntegrityBy

Mohamed Hossany Negm

Senior Technical Support Specialist and MarketingMaster’s Degree, Poultry Science, very good

TEL:00201123223874

Faculty of Agriculture Zagazig University

GIT is a very complex organ

(supports Metabolism, Growth and

Performance).

GIT performs two basic functions:

- Acquisition and assimilation of nutrients.

- Maintenance of a barrier of protection against

microbial and viral infections.

Consequently; GIT development

and health is the key to productivity.

Factors influence performances of GIT*Intestinal health.

*Nutrition and feed ingredients.

*Microflora equilibrium.

GIT microflora plays a very

important role in absorption and

nutrient availability.

For the next decade, microflora

management will be the “buzz

word’’ for poultry intestine

Anatomy and physiology of the poultry GIT

is sufficiently different when compared to

monogastric mammals.

Nutritional requirements (much higher)

Microbial imbalance poorer performances

(disrupt digestion and absorption).

Consequently; It is necessary to

design appropriate nutritional and

feeding programs and additive

strategies.

I- Immunosuppressive Factors:

A-Feed: mycotoxins, antibiotics, pesticides, chemicals, deficiency

in: vitamins,Protein & minerals.

B-Environment: bad ventilation (ammonia and dust- cilliary

apparatus), temp. extremes, air drafts chilling, over heating, relative

humidity.

C-Microorganisms:1-Viral: *Vertically transmitted (CAV, ALV, REV &Reo virus).

*Horizontally transmitted (IBDV, MDV, NDV & AIV).

2-Bacterial: E.coli, MG & MS.

3-Parasitic: Coccidiosis.

Predisposing Factors For GIT

Infections

Predisposing factors (Continue)

II-Management factors:1-System of management (Multiage).

2-Distance to the nearest farm.

3-Distance between houses.

4-Stocking density (over crowding - social stress)

5-Chick quality.

6-Feeders and waterers (amount & quality).

7-Poor biosecurity (Rodents, Insects, Animals,

personnel).

8- Litter (dry or wet).

9-Duration between successive cycles.

10-Vaccination and medication.

Avian Clostridial infectionsClostridia organisms are straight or slightly curved gram positive

anaerobic rods with rounded ends, with oval subterminal spores

Disease conditions associated with Clostridialinfections:1- Gangrenous Dermatitis (C.septicum, C.perfrinens A).

2- Botulism (C. botulinum type C toxigenic group).3- Enteric infections:

a-Ulcerative Enteritis (C. colinum).

b-Necrotic Enteritis (Clostridium. Perfringens A and C).

N.B.: C. Perfringens causes cholangiohepatitis in chickens, turkey tail

cellulitis, gizzard erosions in layer pullets, and navel infections in

neonatal chicks. C. difficile causes enterotoxemia in young ostrich

chicks. C. piliforme causes Tyzzer’s disease in parrots.

Gangrenous Dermatitis(Gangrenous Cellulitis, Necrotic Dermatitis, Gangrenous

Dermatomyositis, Avian Malignant Edema, Wing Rot, Blue Wing Disease)Etiology:

C.septicum, C.perfrinens type A, Candida albicans and Staphylococcus

aureus either singly or in combination.

Epizootiology:

*Chickens, turkeys are susceptible.

*Clostridia are distributed in soil, feces, dust, contaminated litter or feed

and intestinal contents.

*Staphylococci are ubiquitous and common inhabitants of skin and

mucus membranes of poultry.

*Gangrenous dermatitis is believed to occur as a sequel to disease

produced by other infectious agents such as IBDV,CAIV, REV,and

Adenovirus (inclusion body hepatitis).

Extensive blood-tinged edema

with or without gas

(emphysema), is present

beneath affected skin.

Discolored muscle and

serosanguinous fluid

expanding underlying dermis

Spontaneous

separation of

epidermis

revealing

edematous,

hyperemic, acutely

inflamed dermis.

Dark moist areas of skin, usually devoid of

feathers, overlaying wings, breast abdomen,

or legs.

Underlying musculature is discolored gray or tan, and may

contain edema and gas between muscle bundles . serosanguinous

fluid in subcutaneous tissue

Clinical and Pathological Picture

Mortality ranges from 1—60% .

Diagnosis and Differential Diagnosis of Gangrenous

Dermatitis1-CPP.

2-Isolation of of the causative agent(s).

Planter pododermatitis of

turkeys

Squamous cell carcinoma

Vitamin B

complex

(Biotin

deficiency)

Vitamin B

complex

(pantothenic acid

deficiency)

A- Treatment:

Chlortetracycline, oxytetracycline , erythromycin , penicillin ,

or copper sulfate in the water and chlortetracycline or

furoxone in the feed.

B- Water acidification, with citric and proprionic acid, have

been used to reduce but not eliminate mortality.

C- Thorough cleaning and disinfection of the house and

floor.

D- Improve litter condition,

E- Reduce moisture and bacterial levels in the environment.

F- Minimize trauma.

Prevention and control of

Gangrenous Dermatitis

Etiology:C. botulinum (type C toxigenic group).

Epizootiology:GIT obligate parasites. Their spores commonly

found in and around poultry farms.

Resistance of spores to inactivation, favor

spread of this organism.

Botulism (Limberneck Paralysis)

Diarrhea with

excess urates

in the loose

droppings.

Paralytic signs

progress

cranially from

the legs to

include wings

(drooped

wings), neck

(Limberneck),

and eyelids

(birds appear

comatose).

Gasping

when birds

are handled.

Infected birds

are found

setting and are

reluctant to

move.

Death results from cardiac and respiratory failure.

Lack gross or microscopic lesions

Clinical and Pathological Picture

Marek’s disease

Diagnosis and differential

Diagnosis:

*Clinical signs.

*Lack of gross or microscopic lesions.

*Definitive diagnosis by detection of

toxin in serum, crop, or gastrointestinal

washings from morbid birds by mouse

bioassay (mice inoculated with specific

antitoxin will be protected).

Differentiated from:

1- Marek’s disease.

2- Drug and chemical toxicity.

A-Treatment:

1-Vitamins AD3E, Bacitracin (100 g/ton in

feed), Streptomycin (1g/L in water),

Chlortetracycline.

2- Specific antitoxin (Valuable birds).

B-Prompt disposal of dead birds (Cannibalism)

C-Thorough cleaning and disinfection of

litter (calcium hypochlorite, iodophor, or

formalin).

D-Fly control (Toxic maggots).

Prevention and Control of Botulism

Ulcerative Enteritis (UE)

Necrotic Enteritis (NE)

Intestinal Pathological lesions(In some cases M.O. are relatively innocuous unless there are

cofactors (Dietary changes, Stress, Coccidiosis,

Immunosuppressive infections)

Enteric clostredial infections

Toxins

Ulcerative enteritis (UE)Acute bacterial infection of young chickens,

turkeys, quail - sudden onset - rapidly increasing

mortality.

Etiology:

C. colinum

Epizootiology:

Wide range of species - Accompany or follow

Coccidiosis, CIA, IBD or Stress - Transmission

through droppings – Infection via ingestion of

contaminated feed, water or litter - Chronic

carriers.

Clinical and Pathological Picture of UE

•Acute disease: No signs- Sudden death .

• Less acute: Listlessness, humping up,

partly closed eyes, ruffled feathers with

severe emaciation.

•Mortality: in young quail may be as high

as 100% in a matter of a few days. Chicken

losses typically range from 2—10%.

LiverYellow mottling to large irregular

yellow areas along the edges.

CecaUlcers in ceca have a central depression filled with dark

staining material coalescing together forming large

diphtheritic patches that can not be rinsed off. Perforation of

ulcers frequently occurs, resulting in peritonitis and intestinal

adhesions.

IntestineSmall yellow foci with

hemorrhagic borders in intestinal

serosa and mucosa, increase in

size, coalescing to form large

necrotic diphtheritic patches.

Ulcerations may be extensive

enough to erode through the

intestinal wall, perforating the

intestines resulting in peritonitis

UE

SpleenCongested,

Enlarged, Hgic.

2- Histomoniasis(Enterohepatitis- Blackhead

disease).

1- Coccidiosis

1-CPP.

2-Isolation of of the causative

agent(s): Crushed necrotic liver

tissue between two slides, fixed

by heat, and stained by gram’s

stain. Isolation of C. colinum

from liver or spleen or

ulcerative lesions. FAT, agar gel

immunodiffusion test.

Diagnosis and Differential Diagnosis of UE

Differential Diagnosis :

Diagnosis :

Etiology: Clostridium. perfringens types A and C

producing alpha toxin (C. perfringens A and C) and beta

toxins (C. perfringens C).Epizootiology:GIT obligate parasites. Their spores commonly found in and

around poultry farms.

High levels of barley, fish meal or wheat as well as damage of

the intestinal mucosa (high fiber content or coccidiosis)

predispose for NEN.B.:

Possible vertical transmission of C. perfringens has been shown.

Necrotic enteritis (NE) Enterotoxin may cause serious necrotic lesions in gut

wall, as well as increased mortality and/or liver

condemnation at slaughter.

Flecks of

blood

Hyperemia and

diffuse necrosis

of the mucosa

with multifocal

ulceration

Friable intestine,

distended with gas

with mottling

serosal surface

Fibrinonecrotic

enteritis with formation

of diphtheritic

membrane.

General clinical

signs-Reluctance to

move-Diarrhea-

Acute death

Clinical and Pathological Picture of NE

Coccidia Clostridium Mortality %

None None 0

None Yes 16

E. Acervulina Yes 53

E. necatrix Yes 28

Interaction of Coccidia and NE

(Clostridium perfringens)

Groups

Total mean lesion

score (Day 36-42)

E.

maxima

C.

perfringens

Uninfected Control 0 3.8*

Infected with C. Perfringens 0 10.8

Infected with E. maxima 3.4 6.4*

Infected with E. maxima at 28 days and

Infected with C. Perfringens at 34 days 2.4 13.2

* Mild NE lesions possibly due to clostridia infection from the hatchery or spread from experimentally

infected birds.

Mean coccidial and NE lesion scores (each range 0-4) in birds

given various combinations of clostridia and coccidial infections.

Source R. B. Wiliams et al., 2003, Parasitology Research, 90:19-26)

1-CPP.2-Isolation of C. perfringens

Diagnosis and Differential Diagnosis of NE

Differential Diagnosis :

Diagnosis :

1- Ulcerative enteritis.

2- Eimeria brunetti infection.

1-Veterinary biosecurity and hatchery hygiene

2- Destruction of large numbers of Clostridial

pathogens :

* Using disinfectants.

* Feed decontamination through:Physical destruction of microorganisms(irradiation,

Heating).

Chemical mechanisms(Dietary acid additives).

3-Strengthen resistance of birds.

Prevention and Control of

GIT Clostridial Pathogens

.

.

.

.

* Clean out and disinfect houses thoroughly between crops.

* Improve hygiene in hatcheries and during transportation

* Pick up carcasses as soon as possible to restrict contamination.

* Avoid wet litter.

* Treat litter with acidifiers.

* Increase period between crops.

* Fly control (reduce the risk of toxic maggots).

1- Hygienic measures:1- Veterinary Biosecurity

2- Reduce:

* Proportion of wheat in feed.

* Clostridia contamination (disinfectants on farms).

* Chick placement density.

* Fish-meal in diets.

N.B.:

Severe coccidiosis may exacerbate clinical effects of clostridia infection.

Traditional anticoccidials may directly control NE, and ionophores may control

both coccidiosis and NE.

2- Destruction of large numbers of Clostridial pathogens 2.1- Using Disinfectants and Sanitizers.

2.2- Feed decontamination.

Finally bacteria are

killed in the hostile

environment by

combined effects of

acids, higher water

activity and body

temperature

Non-dissociated

organic acids

penetrate

bacterial cell wall

& disrupt normal

physiology

Rel

easi

ng

Protons (+)(Hydrogen ions) Bacterial internal

pH decrease

Specific mechanism will act to bring pH to

normal level consumes energy and stop growth

Disrupt DNA

synthesis

Physical methods

(but able to re-

infection)

Heat treatment:1-Pelleting (temperatures

of about 70 C; but exposure

time is not sufficient to reduce

significant rates of

contamination).

2-Expander-system (105-

130 C for just a few seconds on

pressure - up to 50 bar).

3-Extruder (115-155 C for

30 seconds result in complete

decontamination).

Irradiation (not

allowed in many countries).

Protected organic acids in a matrix crossing upper digestive system without

denaturation. In intestine, matrix will be emulsified and hydrolysed by liver

and pancreatic secretions and intact acids liberated in non-dissociated form.

Chemical methods

3- Strengthen Resistance of

Birds

Chemotherapy

(antibiotics).

Competative

Exclusion.

ChemotherapyRole of antibiotics in therapy can not be ignored. Growth

promoters has proven useful in stabilizing GIT microflora

and improving general performance of poultry and

preventing some specific intestinal pathologies

However !

✺Tedious and Costly Process (need special factories and

environment, as well as a special well trained and educated

labors).

✺ Residues (withdrawal time).

✺ R-Factor (creation of resistant Salmonellae and Coli-forms

for human beings

✺Problems face production (e.g. yeast mutation, temperature

changes, fermentors…).

1- Subtherapeutic

use (G.P.) or

misuse results in

2- Therapeutic

use affects on

Indigenous gut flora (intestinal

upsets) and persist even after

their cessation (Watkins and Kratzer;

1984).

1- Development of resistant

population of bacteria (So

subsequent use for therapy is

difficult, Ghadban;1999).

2- Negative effect on lactic

acid bacteria (Bougon et al.,1987).

Moreover !

WHO is urging the meat producing

countries around the world to use

“Environmentally Friendly”

alternative methods to control

infectious diseases

American Medical Association

urged that non-therapeutic use in

animals of antimicrobials should be

terminated.

Now many supermarkets

are already selling

antibiotic-free meat

In 2006 all growth promotors already banned in EU.

Many other countries will have to follow this measures to

export to EU. Consequently; Many governments around

the world are banning or severely limiting the use of

antibiotics at non-therapeutic levels.

Alternative methods of

controlling infectious

diseases are looked for

Raising poultry without antibiotics may result in

?

Poor

performance Enteritis

1- Economic problem (Engster et al.; 2002).

2- Animal welfare problem (Norton; 2000).

Competative Exclusion

As early as 1928, Voltera reported that

native micro-flora competitively exclude

bacterial pathogens from the intestinal tract

of poultry.

Accordingly; Competitive exclusion (CE)

is a term describes the protective effect of

natural or native bacterial microflora of

GIT which limits colonization of some

bacterial pathogens.

Day-old-chick prior to acquiring the

normal healthy adult intestinal flora.

Protective adult bacteria

of many different strains

colonise the caecum.

Nigel Horrox (1997) mentioned that the bird’s

microflora is potentially depleted for a period of

time at hatching and following any medication

with an anti-microbial product.

Electron Microscopy of Caecum

Competative Exclusion

First CE products were simply fecal

contents. Researchers have developed

defined mixtures of bacteria or yeasts for

use as CE products.

These products are called:

Probioticsor direct-fed microbial

or CE cultures

Competative Exclusion

Probiotic* Pure culture of one or more living M.O. given in

feed and proliferate in the host-bird’s GIT.

* Implies antagonistic activity and competition for

attachment sites by ensuring that the bird

Maintains beneficial

microbial population in GIT

Competative Exclusion

Improving feed intake and digestionIntestinal bacterial flora takes part in the metabolism of nutrients

and synthesis of vitamins (Nahanshon et al.; 1992 and 1996).

Immune stimulation1- T cell function Enhancement

and increase anti-salmonella

IgM antibodies (Dunhan et al.;

1993).

2- Stimulation of lgA production

(Nahanshon et al.; 1994).

3- Increase macrophages

effectiveness (Goldin and

Gorbach (1984).

Altering metabolism*Increase digestive enzyme activity (amylase, protease and lipase)(Ghadban;1999).

*Decrease bacterial enzyme activity (Reduction in b-glucuronidase )(Coloe et al.;

1984 and 1987 and Jin et al.; 1997).

* Reduce ammonia in the excreta and litter of broilers (Chiang and Hsiem; 1995).

Maintaining

beneficial

microbial

population in

GIT

Microbial antagonisms1- Production of bacteriocins,

organic acids and hydrogen

peroxide (Fuller; 1989).

2- Neutralizes enterotoxins

produced by pathogenic

bacteria (Schwab et al.;

1980)

Competative Exclusion

Usage of probiotics is recommended:1-In newly hatched chicks for rapid

establishment of microflora (to safeguard the

host against enteric bacterial pathogens) (Mead;

2000).

2-In altered intestinal flora due to stress,

usage of antibiotics or coccidiostats (to

restore a protective microflora) (Joan Jeffrey; 1998).

Competative Exclusion

This microbe versus microbe army is

the body's first line of defense that

prevents a range of illnesses. Certainly this might be valuable in decreasing the

use of antibiotics (whose usage may result

in bacterial resistance and formation of

residues in organs and tissues of treated

birds).

Competative Exclusion

Sensitivity of

some GIT

bacteria to low

pH

1 2 3 4 5 6 7 8 9 103.5 4.5 Moulds

Yeasts

Streptococci

C. perfringens

Staphylococci

SalmonellaeE. coli

Lactic acid bacteria

Fuller (1989) concluded that the auspicious effect of

probiotics over the host is due to: 1-Stooping implementation of pathogenic bacteria over intestinal mucus

membranes.

2-Better adhesion of lactic acid bacteria to intestinal epithelium.

Lowering

PH