CLOSTRIDIAL DISEASES

Necrotic enteritis (NE)

NE is found worldwide. Disease is most common in broiler chickens and meat turkeys. Broilers
aged 2-5 weeks are most frequently affected. It is also found regularly in layers, mostly in pullets
and young birds kept on litter.
Cause - is Clostridium perfringens type A or C in large intestine & caeca and
subsequent migration to small intestine where it produces toxins. Alpha toxin produced by Cl.
Perfringens types A and C and beta toxin produced by type C, are believed responsible for
intestinal and mucosal necrosis (characteristic lesion of NE). Both have been detected in feces of
birds with NE Predisposing factors include outbreaks of coccidiosis (mild/subclinical), changes
in diet (> starch and less fibre), anaemia and inadequate cleaning of poultry house, utensils and
equipment.

SPREAD - Clostridium perfringens can be found in feces, soil, dust, contaminated feed and
litter, or intestinal contents. Contaminated feed or litter are the main source of infection. Fish
meal is considered a most likely source of Clostridium perfringens contamination.

Pathogenesis: The presence of the organisms in intestine alone is not sufficient to induce necrotic
enteritis. The following two requirements have been proposed for induction of necrotic enteritis
i. The presence of some factor causing damage to the intestinal mucosa, and ii. Presence of
higher than normal numbers of intestinal Cl. perfringens organisms.
If these two requirements are fulfilled, lesions may develop, often starting at the tips of
the villi. Bacterial cells adhere to damaged epithelium and denuded lamina propria where they
proliferate and induce coagulative necrosis. Attraction and lysis of heterophils granulocytes as
well as further tissue necrosis and bacterial proliferation proceed rapidly.
Alpha-toxin, a necrotizing toxin produced by all Cl. perfringens types, has been assumed as an
important virulence factor. This toxin destroys cell membranes by hydrolysis of membrane
phospholipids. Toxins may also enter blood stream, causing systemic effects and death.

Clinical signs: Acute NE is characterized by increased mortality but few visibly sick birds,
indicating that affected birds die rapidly. The signs include depression, decreased feed intake,
and reluctance to move, ruffled feathers, drooling of saliva and diarrhoea. The mildest form of

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NE induces no visible illness of the birds but is associated with temporarily reduced weight gain,
impaired feed conversion ratio and increased condemnation rates at slaughter due to liver lesions.

Gross lesions - Characteristic gross lesion of NE is enteritis in upper part of small intestine with
necrosis of mucosa in large patches forming pseudomembrane attached to the intestinal mucosa.
The mucosa of caecal pouches are not changed but caecal tonsils and adjoining area of caeca
may be affected, occasionally. The membrane may be white, yellow, green, brown or red. A
yellow or green discolourations is the most commonly found. Birds dying with NE often show a
dark liver with dilated gall bladder, pale kidneys with prominent lobular outlines, and dark & dry
pectoral musculature indicating dehydration. The body condition depends on the course of
disease. Birds dying with acute NE are in good bodily condition. Three main types of liver
lesion may be found. The most common and most characteristic is cholangiohepatitis. Livers
with second type of lesion show light, randomly localized nodules (focal necrosis and
granulomas) of the liver parenchyma, and the third and most rare type is massive liver necrosis
causing a smaller or larger part of a liver lobe to be homogenously discoloured.

Microscopically - characteristic feature is an aggregation of large, Gram-positive, rod-shaped

bacteria surrounded by necrotic tissue delineated from viable tissue by a zone of granulocytic
infiltration containing pyknotic cell nuclei. Lesions are usually located within the lamina propria
of gut mucosa. The least extensive lesions are limited to focal necroses comprising some epithe-
lial and stromal cells on villus tips. The deepest lesions may even affect the muscular wall of
gut.

Diagnosis - Gross and microscopic lesions of the disease are adequate. In doubtful cases,
bacteriology, histopathology and examination for coccidian will be helpful.
Differential Diagnosis - The similar lesions producing disease like coccidiosis and ulcerative
enteritis should be considered.

Hemorrhagic Enteritis or Ulcerative Enteritis (UE)

It is an bacterial infection of young chickens and turkeys characterised by sudden onset
and rapidly increasing mortality. The disease enzootic was first seen in quail, therefore
called’Quail’s disease’. Now it has been established that many avian species are susceptible.

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Therefore, the earliest name has been replaced by ‘ulcerative enteritis’.
Cause - It is caused by Clostridium colinum, an anaerobic spore-forming bacterium
with fastidious growth requirements. The organism is ubiquitous in nature and is excreted in
large numbers in feces of affected stock, which are an important source of infection. The spores
result in persistent contamination of premises after an outbreak. Influencing factors appear to
play an important part in the production of disease include: Coccidiosis, immunosuppressive
factors such as infectious bursal disease, chicken aneamia virus and others, and overcrowding
and inadequate hygiene. Keeping chickens on wire floors has been reported to be preventive.
CLINICAL SIGNS - In acute disease, there may be increased
mortality without any obvious signs. In other case, signs may include depression, huddling, with
ruffled feathers, anorexia and watery droppings. Mortality in chickens vary from 2% to as high
as 12%. Gross lesions - Birds dying with acute disease
show good condition and may have feed in the crop. More protracted disease may lead to
emaciation. The most important lesions are found in intestine, liver and spleen. First, there are
small, circular mucosal ulcers affecting the small intestine, caeca and upper large intestine. The
ulcers may penetrate as deep as the serosa, which may become perforated and result in
peritonitis. The ulcers may coalesce to form large areas with a pseudomembrane. Small ulcers
have a hemorrhagic border, which may be seen on serosal and mucosal surfaces. A hemorrhagic
border is less frequently found in larger lesions. In liver, there are yellowish to grey necrotic
lesions of varying size. Spleen is enlarged and hemorrhagic.
Microscopical lesions - Intestinal ulcers consist of small haemorrhagic and necrotic areas, often
with clumps of Gram(+) bacteria. The ulcers involve villi and extend into the submucosa. The
ulcers sometimes reach as deep as muscular coat and serosa. Affected tissue is surrounded by
granulocytes and mononuclear inflammatory cells. Liver lesions consist of multifocal foci of
coagulative necrosis that are often poorly demarcated and with minimal inflammatory reaction.
Gram-positive bacteria are occasionally found within the necrotic foci.
DIAGNOSIS - Gross lesions are often sufficient to make presumptive diagnosis. A definitive
diagnosis requires detection of causative bacteria in affected birds. Liver is the most suitable
organ for isolating Cl. colinum. Slide smears of liver and spleen tissues may be Gram-stained for
detection of Gram-positive bacilli and subterminal and free spores. The organism can be

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demonstrated from a direct fluorescent antibody test in cryostat sections of liver and intestine.
Differential diagnosis should be made for necrotic enteritis, coccidiosis and histomoniasis.

Gangrenous dermatitis (Malignant edema)

Disease occurs worldwide in chickens and turkeys. The condition is precipitated by
various detrimental microbial, nutritional and environmental factors. The disease is character
rized by a sudden onset, sharp increase in mortality, and gangrenous necrosis of the skin over the
wings, thighs, breast, and head. It occurs sporadically in chickens 4-16 wk old, affects broiler
and layer replacement stocks, and occasionally causes outbreaks in turkeys.
CAUSE - It may be associated with various aerobic and anaerobic bacteria; however Clostridium
septicum, Clostridium perfringens type A, and Staphylococcus aureus, either singly or in combi
nation are most often involved. Combined infections are often more severe. Young chicks
immunosuppressed by infectious bursal disease or chick anemia virus are predisposed. Disease
may occur secondary to avian adenovirus or reticuloendothelial virus infections as well. Skin
lesions due to trauma, wet litter, picking, or treading wounds may provide entry sites for
causative bacteria. Systemic effects arise from invading bacteria and their elaborated exotoxins.
CLINICAL SIGNS - First sign is usually a sudden dramatic increase in mortality in affected
flock. Overall mortality is 10-60%. Affected chickens are extremely depressed, lethargic, and
prostrate, and die within 8-24 hr. Red to black patches of moist, gangrenous skin are seen over
the breast, abdomen, wing tips, or thighs. Feather loss or sloughing of the epidermis is common.
When clostridial infection occurs, palpation of the affected areas often reveals crepitation due to
gas bubbles in the subcutis and musculature.
Gross Lesions - At necropsy, there is accumulation of gaseous, serosanguineous fluid in
subcutis, and musculature has a pale cooked appearance. Lliver and spleen are enlarged and may
contain infarcts or pale focal areas of necrosis. Kidneys are usually swollen, and the lungs may
be congested and edematous or necrotic. Atrophy of the bursa of Fabricius may be found in birds
that were exposed to infectious bursal disease virus in the first few weeks after hatching.

Histopathologic demonstration of gangrenous necrosis with numerous coccoid bacteria or large,

gram(+) rods with or without spores in affected tissues is sufficient to confirm a clinical
diagnosis. Diagnosis - Isolation of the etiologic agent (Cl. Septicum, Cl.
Perfringens, or Staphylococcus aureus), together with the history and clinical findings.

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Diff. Diagnosis - Disease should be differentiated from exudative diathesis, fowl pox, Squamous
cell carcinoma of skin of chickens.

Avian tuberculosis - Avian tuberculosis occurs throughout the world in many avian and some
mammalian species and in domestic poultry it is generally seen in mature stock kept in
conditions of poor management. It usually runs a protracted course, causing reduction in
condition, reduced egg production and eventually death. Although loss in a flock is intermittent it
is invariably in adult fowls and this, together with the culling of unthrifty birds and the
depression in egg production, can cause serious economic loss. The infection is of importance
also because the disease occurs in wild birds, pigs, rabbits and mink.

EPIDEMIOLOGY - Cause - Mycobacterium avium is the name given to a complex group of

mycobacterial organisms that, according to current taxonomy, consists of four subtypes, M.
avium subsp. avium consists of three serotypes (1, 2 and 3) and several genotypes: this subtype is
fully virulent for birds and small terrestrial mammals. M. avium subsp. Paratuber
culosis consists of a number of genotypes and affects ruminants and other animals.
These mycobacteria are acid- and alcohol-fast whcn stained by the Ziehl - Neelsen method and
the organisms often appear beaded. Of this large number of serotypes it is types I, 2 and 3 that
are most virulent and are mainly responsible for the disease in poultry. Even among these
serotypes there is considerable variation in virulence.

M. avium, compared with other mycobacteria, is relatively resistant to antimicrobials and

relatively resistant to a number of disinfectants but is sensitive to ionic detergents. Outside the
body it can survive for many years but the unprotected organism is killed by direct sunlight and
within the carcass the organism survives for no more than a few weeks.

HOSTS - It is probable that all species of bird can be infected but susceptibility among domestic
species seems to be in the following order: chickens, ducks, geese and, least susceptible, turkeys,
in which it is relatively uncommon. The disease is observed most commonly in older poultry
because of the greater opportunity for infection with age and the generally long incubation
period. However, occasionally, heavy losses may occur in pullets on multiage sites where the
infection is endemic and the standards of hygiene poor. Game birds, particularly

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pheasants, are also susceptible. Some birds kept in zoological gardens seem to be prone to
tuberculosis, perhaps because of the difficulty in adequately cleaning and disinfecting pens. Cage
birds may also succumb to avian tuberculosis but tuberculosis in parrots and canaries may also
be caused by M. bovis or M. tuberculosis. Among mammals M. avium can cause progressive
disease in swine, rabbits and mink and can cause sensitivity in cattle to the skin tuberculin rest.

SPREAD - In the transmission of infection the most important source of the organism is the
infected host, including domestic poultry, game birds and per or wild birds. Next in importance,
because of the prolonged survival of Al avium outside the body of the host, are items
contaminated with the droppings and excrement of such birds. These commonly include litter,
contaminated pens and pasture, equipment and implements that come into contact with infected
hosts, and the hands, feet and clothing of attendants.

INFUENCING FACTORS - The infections are worldwide but disease varies between and
within countries. In domestic poultry lack of hygiene in management and the age of the birds
influence the appearance of the disease since the organism is highly resistant in the environment
and within the host is generally associated with a long incubation period.

Clinical Signs - may be prolonged over a period of weeks or months before death. There is
generally progressive but slow loss of condition and accompanying loss of energy and increasing
lethargy. Although the appetite usually remains good, there is eventually gross emaciation with
marked atrophy of the sternal muscles, with the ‘keel’ becoming prominent or even ‘knife-
edged’. The face and comb become pale and sometimes jaundiced and the comb is shrunken and
often there is persistent diarrhoea with soiling of the tail feathers.

Occasionally, birds may die suddenly in good bodily condition and yet show advanced lesions of
tuberculosis. In such eases rupture of the affected liver or spleen with consequent internal
haemorrhage is often the precipitating cause of death.

Gross lesions - in the chicken, are most commonly seen in the intestines, liver, spleen and bone
marrow but may be found in any organ or tissue. Irrespective of the organ involved, the lesions
are typical tubercular granulomata. They are irregular, grey-white nodules, varying in size from
pinpoint to large masses of coalescing tubercular material. ‘When cut through, the nodules are

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firm and caseous and the centres may be a pale yellow colour, particularly those from the bone
marrow. Those in the liver and intestine may show bile staining. llae liver and spleen are often
grossly enlarged and occasionally rupture, resulting in blood in the body cavity and sudden
death. The wall of the intestine is invariably studded with similar lesions, varying in size from a
millimetre to several centimetres in diameter. They usually involve the whole thickness of the
intestinal wall and eventually ulcerate into the lumen of the intestine, with consequent discharge
of bacilli and probably constituting the major source of infection within the droppings.

DIAGNOSIS - The clinical signs and gross lesions are strongly indicative of avian tuberculosis
and the demonstration of acid/alcohol-fast tubercle bacilli in lesions or sections is supportive of
this. There is seldom any difficulty in demonstrating the organisms, which are often present in
very large numbers, particularly in young lesions and those from the bone marrow. Cultural
examination, or even chick inoculation of suspect material, may be necessary when organisms
are few or for isolation and identification of the causal agent. The agent can also be identified by
DNA techniques.

Immunological tests are also of value in the recognition of infected birds during life. They
include the tuberculin test, an agglutination test and ELISA. The tuberculin test in the fowl
consists of injection of 0.05-0.1 mL of avian tuberculin into one wattle using a needle about 1 cm
long and of 25 gauge. The other wattle remains uninjected as the control. When testing a flock it
is usual to inject the tuberculin into the wattle on the same side for each bird. The needle is
introduced at the lower edge of the wattle and is directed upwards into the centre. The test is read
48 h after the injection of tuberculin, although some positive reactions may be observed sooner
than this. The test is read by palpating the two wattles simultaneously between the first finger
and thumb of each hand. A positive reaction is recognized by a hot, soft, edematous swelling of
the injected wattle, which maybe twice the size of the uninjected one or even larger. Most
uninfected birds will show no reaction in the injected wattle and occasional small, firm, pea-like
swellings can usually be ignored. The accuracy of the test, relative to gross lesions seen at
necropsy, in detecting infected birds is about 80%. However, birds in an advanced stage of
infection may give no reaction. It is possible, however, that such birds would be thin or
emaciated on handling during the testing of a flock and thus arouse suspicion of tuberculosis.

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DIFFERENTIAL DIAGNOSIS - In differential diagnosis, at necropsy, most difficulty might
be in differentiation from neoplasia but the simple enucleation of tubercular lesions from the
surrounding tissues and the demonstration of typical acid fast organisms should be adequate.