Anthrax

Anthrax is an acute serious highly infectious zoonotic disease of herbivorous caused

by the spore forming gram positive Bacillus anthracis. This disease is also known as
splenic fever due to the fact that there is extensive enlargement of the spleen
(Splenomegaly) due to this infection

Etiology: Large Gram positive, sporulating nonmotile, weakly hæmolytic; bacteria

with square straight ends, encapsulated in vivo, produces long chains. - Pathogenic
to herbivores, man, lab animals. - Habitat: Parasitic; persists in “cursed” fields. -
Sporulation only in aerobic conditions.

Pathogenic strains have a capsule and three virulent toxins;* the capsule that aids in
resistance to phagocytosis, * the complex toxin consists of a 1.lethal factor.
2. Oedema factor. 3. Protective antigen.

 Protective antigen acts as the binding activation part for both factors.

 Oedema factor------once it entered the cell following binding to protective

antigen-----increase in cyclic AMP ----- disturbance of water homeostasis----
fluid accumulation (oedema).

 Lethal toxin binding to protective antigen causes------- death of macrophages,

neutrophils, dendritic cells, some epithelial and endothelial cells.

 In naturally occurring disease, local effect of the complex toxin includes

swelling and darkening of tissues due to oedema and necrosis.

 When septicemia occurs----generalized increase in vascular permeability and

extensive haemorrhages-------shock and death.

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Occurrence: Worldwide in distribution; area prevalence varies according to soil,
climate and efforts put forward to suppress its occurrence.

In developed countries there is simultaneous occurrence of multicentric foci of

infection. Many cases of sudden death occur in areas showing recent appropriate
climatic conditions and in which anthrax has occurred long time ago (about 30 years).

In tropical and subtropical areas with high annual rainfall the infection persists in the
soil, so frequent serious outbreaks of anthrax are common each summer.

In temperate and cool climates only sporadic outbreaks of soil born infection is
observed. Outbreaks are small with small number of animals affected through
accidental ingestion of contaminated bone meal or pasture.

Transmission:

 Ingestion of contaminated food or water, injury to the digestive mucosa may

facilitate infection, but there is little doubt that infection can take place through
intact mucosa.
 Inhalation Of miner importance in animals, in man "Wool sorters disease" is
due to inhalation of anthrax spores in wool and hair industries.
 Through skin abrasions, biting flies and other insects have been found to
harbor anthrax organisms, it is a mechanical transmission.
Animals affected: All vertebrates; but it is more common in herbivorous animals as
cattle and sheep, less frequent in goats and horses, swine dogs and cats are
relatively resistant. Humans are intermediates between highly susceptible and
relatively resistant vertebrates.

Environmental factors: A soil borne infection outbreaks (Anthrax years)always occur

after a major climate change (heavy rain after a prolonged droughts, or dry summer
months after prolonged rain, and always in warm weather when environmental
temperature is more than 15°C. Spores are present in high concentration in wet soils
they remain suspended in in standing water with further concentration on the soil
surface as water evaporates. Grazing on tough scratchy feed in dry times, results in
abrasions of the oral mucosa and confined grazing on heavily contaminated areas
around water holes.

Zoonotic potential: More than 95% of human anthrax cases take the cutaneous form
and result from handling infected carcasses or hides, hair, meat or bones from such
carcasses. Protection for veterinarians and other animal handlers involves wearing
gloves, and other protective clothing when handling specimens from suspected
anthrax carcasses and never rubbing the face or eyes. The risk of gastrointestinal
anthrax may arise if individuals eat meat from animals infected with anthrax.
The risk of inhaling infectious doses becomes significant in occupations involving the
processing of animal byproducts for manufacturing goods (industrial anthrax). These
include the tanning, woolen, animal hair, carpet, bone processing, and other such
industries, where the potential for aerosolisation of substantial numbers of
spores increase the risk of exposure to infectious doses.

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Pathogenesis: Ingestion of spores infection through intact mucosa-
scratched mucosa- epithelium around erupted teeth resist phagocytosis
proliferation in regional draining lymph nodes lymphatic vessel and blood
circulation septicaemia invasion of all body tissues production
of lethal and oedema toxins edema and tissue damage renal failure
and shock and terminal anoxia death.

Clinical signs:

 Cattle and sheep: Two forms are common in these animals;

1. Peracute form; *most common in the in the beginning of the outbreak. *Animals
are found dead without any premonitory signs, but fever, muscle tremor, dyspnea
and mucosal congestion may be observed.* Course of the disease is about 1-2
hours. *After death, blood discharged from mouth, nostrils, anus or vulva is
common.
2. Acute form; *runs a course of about 48 hours. * There is severe depression and
listlessness in the beginning of the disease. * Sometimes preceded by a short
course of excitement. * High fever, up to 42°C. * Rapid and deep respiration and
increased heart rate. * Congested and haemorrhagic mucosae.* Anorexia and
ruminal stasis.* Pregnant cows may abort. * In milking cows there is severe
decrease in milk production, milk may be bloody or deep yellow in colour. *
Alimentary tract involvement is usual with diarrhea and dysentery. * Local oedema
of the tongue, throat, sternum, perineum and flanks may occur.
Horses: Anthrax is always acute in equine, sometimes it varies according to the
mode of infection
Through ingestion; there is septicemia with enteritis and colic.
Through insect bite; there is hot, painful, oedematous, subcutaneous swellings
appear about the throat, lower neck, floor of the thorax and abdomen, prepuce,
and mammary gland. There is high fever and severe depression with dyspnea
due to oedematous swelling of the throat or colic due to intestinal irritation. Course
of the disease is usually from 48.96 hours.

Diagnosis: 1. direct detection of B. anthracis; in dead or living animal showing

clinical signs:

 Blood and / or oedematous fluid are collected with a syringe from

peripheral blood (ear or tail vein) or subcutaneous oedema avoiding
contamination of the environment. Smears are prepared in the laboratory,
dried and immediately fixed on burner. In a dead animal blood is
collected from ear vein by pricking the area with a needle, making blood
films with immediate fixation on the flame of an alcoholic burner near the
animal in the field, if blood did not appear, cut the ear and make smears
from the cut end avoiding contamination of surroundings with blood.
Punctured or cut ear should be wrapped with goose immersed in xylole or
phenol.
 Fixed slides are then send to the nearest laboratory with caution to be
stained with Gram and capsule stains; examined by a highly qualified
person for the presence of capsulated, large Gram positive bacilli.
Visualisation of the encapsulated bacilli, usually in large numbers, in a
blood smear stained with polychrome methylene blue (McFadyean
reaction) is fully diagnostic. However if the animal has been dead more
than 24 hours, the capsule may be difficult to detect.

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2. Culture: Bacillus anthracis is readily isolated in relatively high numbers from
blood or tissues of a recently dead animal that died of anthrax, and colony
morphology of B. anthracis is quite characteristic after overnight incubation on blood
agar. The colony is relatively large, measuring approximately 0.3–0.5 cm in diameter.
It is grey-white to white, non-haemolytic with a rough, ground-glass appearance..

Colonies of Bacillus cereus on the left; colonies of Bacillus anthracis on the right. B.
cereus colonies are larger, more mucoid, and this strain exhibits a slight zone of
hemolysis on blood agar.

Colonies with ground glass appearance

Tailing and prominent wisps of growth trailing back toward the parent colony, all in the same direction,
are sometimes seen. This characteristic has been described as a ‘medusa head’ or ‘curled hair’
appearance.

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 3. Immunological detection and diagnosis
Ascoli test
Ascoli (1911) for the detection of thermostable anthrax antigen in animal tissue being
used for by-products (hides, wool). This uses antiserum raised in rabbits to produce a
precipitin reaction. The test lacks high specificity.
To perform the Ascoli test, put approximately 2 g of sample in 5 ml of saline
containing 1/100 final
concentration of acetic acid and boil for 5 minutes. The resultant solution is cooled
and filtered through filter paper. A few drops of rabbit B. anthracis antiserum are
placed in a small test tube. The filtrate from the previous step is gently layered over
the top of the antiserum. A positive test is the formation of a
visible precipitin band in less than 15 minutes. Positive and negative control
specimen suspensions should be included.

Precipitation line

Necropsy findings:

1. Striking absence of rigor mortis.

2. Carcass undergoes gaseous decomposition.

3. All natural orifices usually exude dark, tarry blood which does not clot.

Any case of sudden death should be considered as anthrax unless proven otherwise.
The carcass should not be open and subjected to post mortem examination to avoid
sporulation of B. anthracis and long contamination of environment with the highly
resistant spores.

When the carcass is opened under legally controlled circumstances we may found the
following pathological lesions; *failure of blood clotting, *ecchymotic haemorrhages,
*blood stained serous fluids in the body cavities, *severe enteritis and Splenomegaly;
the enlarged spleen is soft looking like a "blackberry jam". *Subcutaneous swellings
contain gelatinous material. *Enlargement of local lymph nodes is an important feature
of anthrax in horses.

Treatment: Early feverish stage of the disease may be treated by:

1. Penicillin- 20000 Unit / Kg BWT.

2. Streptomycin (8-10 g/ day in 2 doses IM for cattle is much more effective than
penicillin).

3. Oxytetracycline (5mg /kg bwt / day; parenteraly proved superior to penicillin.

Control: 1. When an outbreak occurred; *place the farm in quarantine;* destroy all the
discharges and cadavers; *vaccination of survivors; prohibition of milk and meat

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movement in infected herds during the quarantine period to avoid entering infection to
the human food chain. *All suspected cases and in-contact animals must be
segregated until cases cease and for two weeks thereafter. *The administration of
hyper-immune serum or single dose of long acting tetracycline or penicillin to in
contact animals in quarantine may prevent further losses.

When the disease occur in a previously clean area; all in contact animals should be
either treated with hyper immune serum or vaccinated

Disposal of infected materials: Infected carcass should not be openedbut immediately

burned or buried, together with bedding and soil contaminated by discharges.
Carcasses should be buried at least 2 meters deep with an ample supply of quicklime
added.

Disinfection: of bone meals, premises, hides, fertilizers, wool and hair require special
care. Before sporulation takes place ordinary disinfectant or heat (60C for few
minutes) can be used to kill vegetative forms, it is satisfactory when necropsy room or
abattoirs are contaminated. When spore formation occur, within few hours of
exposure to air; disinfection is almost impossible by ordinary means. Strong
disinfectants such as;

 5% Lysol- in contact with the spore for two days.

 Strong solutions of formaldehyde or NaOH (5-10%) are most effective.
 Per acetic acid (3% solution) is an effective sporicide, if applied to the
soil( 8L / m2) is an effective sterilant.
 Infected clothing should be sterilized by socking in 10% formaldehyde.
 Hides, wool and mohair are sterilized commercially by gamma irradiation
from a cobalt source.
Immunization: It is extensively used and many types of vaccines are available.

a. Living attenuated strains of the organism with low virulence but capable of
sporulation is the most successful; they have a disadvantage of producing the
disease in certain species. Saponin or saturated saline solution is put with the
vaccine to delay the absorption.
b. Stern avirulent spore vaccine; has overcome the risk of causing anthrax by
vaccination and produces a strong immunity which lasts for atleast 26 months
in sheep.
Milk of vaccinated cows should be discarded for 72 hours after vaccination, and
withheld from slaughtering for 45 days.