Genus Brucella has 6 species namely:
B. abortus, B. canis, B. melitensis, B. ovis, B. suis except B.
neotome (important pathogens of domestic animals and man)
Brucellosis = disease cause of this bacterial pathogen
Infections = reproductive organs and reticuloendothelial tissues
Abortion = in cattle, sheep, goat because of the lesions of the
female reproductive tract and of the placenta and fetus.
Transmission = directly or indirectly from infected animls to
humans (there is a zoonotic importance because of this)
Brucella melitensis = first member of the genus to be isolated by
David Bruce in 1887
Source = from the spleens of patients who died in Mediterranean
or Malta fever
Brucella abortus = the second member was isolated by Fredrick
Bang in 1897
Source = aborted bovine fetus
Brucella suis = isolated by Jacob Traum in 1914
Source = aborted piglet
B. Ovis and B. canis = more highly host adapted and they were
isolated in 1956 and 1969 respectively
Morphological and cultural features of the 6 members = not
sufficiently characteristic to differentiate. Nor the host from
which the organism is isolated . But each species have a
predilection site for certain hosts.
*Isolation of a Gram-negative rod from a suspected case of
brucellosis requires reliance on the results of the laboratory tests.
Brucella abortus
(Bacillus abortus, Bang’s bacillus)
Affected animal = cattle population (absent in countries where
eradication programs have been actively pursued)
Morphology and Staining reaction
= short rod / a coccobacillus (0.5 to 0.7 µm by 0.6-1.5 µm.)
=rods are short that are mistaken to be a cocci
=arranged singly, although in cultures appear to be in short
chains
= a facultative intracellular bacterium
= found in clumps and smears made from exudates
=Gram negative and stains with ordinary stains although with
some difficulty
=not acid fast
=Can resist decolorization with some mild acids.
Koster stain = which the organism stains deep red
Other characteristics = nonmotile, doesn’t form spore, doesn’t
have a well-developed capsule (poorly developed capsules have
been demonstrated on freshly isolated strains using special
stains)
Cultural and biological features
=aerobic growth (many strains require increased tension of CO2
for growth especially on primary isolations
= is catalase and oxidase positive and usually produce H2S from
protein or peptides rich in amino acids containing sulfur
=produces urease
=8 biotypes are currently recognized by dye sensitivity test, CO2
requirement, H2S production, and presence of A or M Antigen.
= not hemolytic, doesn’t liquefy gelatin, doesn’t produce acid
from glucose
= It has a characteristic substrate oxidation pattern
Growth = enhanced by serum or blood
Tool for primary isolation = complex media (serum dextrose
agar or Albimi Brucella agar or broth)
Antibiotics = they are sometimes added to these basal media to
inhibit contaminants in specimens such as milk and vaginal
discharge.
Primary isolation
= they are slow-growing and barely visible at 48 hours
= reach maximum size after 5- 7 days at the optimum growth
temperature (37oC)
= may be on a smooth type characterized by a round convex
colonies with an entire edge or they may be associated by rough
type colony in which they have large flat colonies with a dull
granular appearance.
=growth is sparse in fluid media and appear as a faint clouding.
Bacteriophage Typing
Bacteriophage = often used for identifying and typing cultures of
Brucella sp.
Tb, Wb, Fi, Bk2, and R strains = the phages currently in routine
Rotuine Test Dilution (RTD) = dilution of the phage; the
minimum number of phages that will produce confluent lysis on
a lawn of the propagating strain
Phage typing = is rapid and very useful tool of identifying B.
abortus
Antigen
cell wall of B. abortus = consists of an outer layer of
lipopolysaccharide protein on which polysaccharide chains are
exposed
Polysaccharide chains = carries the major antigen (A and M)
involved in the agglutination reaction
cell wall proteins have been groouped into three = 88,000-
94,000. 95,000-40,000 (porins); and 25,000-30,000
Epizootiology and Pathogenesis
B. abortus = an obligate facultative intracellular parasite of cattle
and some wild ruminants; they are not very resistant to sunlight
and drying and therefore survives better in winter than summer;
can survive and grow in host macrophages and epithelial cells
Transmission = ingestion of contaminated discharges or feed;
venereal transmission is possible but unusual; intramammary
and congenital transmission also occur
Other infected animals = horses, sheeps, chickens, and dogs
survives = in milk
destroys = pasteurization
Large dairy herds = more likely to be infected with a high
prevalence of infection within the herd. the greater level of
intensivism and opportunity for transmission within large herds
together with the greater likelihood of introduction of a latently
infected animals are some of the factors that contributes to the
higher risk of infection in large herds
factor of transmission = common pastures, contaminations of
streams, activities of carrion feeders
Brucella infection in free-living animals = bison, moose and elk
(comingling with this animals in cattle enhances the possibility
of cross-infection)
Prepubescent animals = resistant to infection
Heifer calves = can be infected early in life either in utero from
infected dams or by ingestion of contaminated milk
(susceptibility increases with sexual development and
pregnancy)
Latent infection = cannot be detected by serological test in
calves exposed in this way
Male cattle = resistant to infection but infection of the
epididymis and testicle has been recorded (Bulls can transmit the
infection in their semen)
Main portals of entry = oral mucosa (milk), nasopharynx, and
conjunctiva of cattle as well as the genital tract in both bulls and
cows
experiimental conditions = organisms has been shown to
penetrate the unbroken skin of guniea pig and cattle. after
penetrationof the host, B. abortus initially localizes in the
regional lymph node and then enters to the blood stream.
bacteremic phase = results in dissemination of the organisms to
the udder, uterus, and associated nodes.
B. abortus = has an unusal resistance to intraleukocytic killing;
proliferates massively in cells with high levels of erythrithol
(found in genital tract in male and pregnant female)
*it penetrates the epithelial cells of the chorion and proliferates,
producing placentitis. *Endometritis is produced with ulceration
of the epithelial lining of the uterus.
*Lesions of the fetus include edema and congestion of the lungs
along with hemorrhages of the epicardium and splenic capsule.
*fetal death occurs (it is unclear wheter endotoxin of the
organism or to interference with placental function)
*inflammation of the membranes(this interference with
thecirculation of the fetus may explain why abortion occurs)
*organisms found in the stomach and lungs of aborted fetuses
*abortion occur late in pregnancy
*organism is present in large numbers in uterine discharges for a
few days but is then gradually cleared from the reproductive
tract (after calving or abortion)
*infection is maintained in the reticuloendothelial tissue and the
udder
Milk = source of infection for calves and also humans (shed in
large numbers)
*most infected animals are remain carriers for life and shed the
organisms in uterine exudates and in milk after each calving
*abortion are usually seen only during the first pregnancy after
infection (placenta is infected during each pregnancy)
*organisms are seen in the lymph nodes of the digestive tract
and in the spleen in the infected cattle.
*it has been isolated from the blood and from hygromas of the
knee (shows high correlation with brucella abortion in African
cattle)
B. abortus in horses = less frequency than in cattle; frequently
localizes in bursae, joints, or tendon sheath and has been found
in poll evil and fistulous withers, supra-atlantal bursitis, and
supraspinous bursitis as well as in lesions of the fetlock and
sternum.
Immunity
*the organisms are so readily escapes the bactericidal effects of
antibody and complement in plasma
protective immunity = depends mainly on cell-mediated
responses in whcich the micorbicidal activity of macrophages is
enhanced following activation by lymphokines from sensitive T-
lymphocytes.
*the organism has been shown to multiply more slowly in the
macrophages of vaccinated calves than in controil animals.
humoral antibodies = corralate poorly with protective immunity
Strain 19 vaccines = cattle vacinated by this have been shown to
be resistant to challenge after antibody titers fall below
detectable levels
IgM agglutinins = first antibodies to appear in the plasma and
their levels reach a peak at about 2 weeks; IgG antibodies appear
later.
IgG1 subclass = infected cattle have high titers of
nonagglutinatingh antibody
pH 3.6 = treatment with acid at this level activates their
agglutinating ability.
strain 19 = IgM antibody levels increase rapidly in calves
vaccinated with this strain; IgG1 antibody levels rise more
slowly, do not attain high high levelsand do not persist. also,
levels of nonagglutinating IgG1 antibodies are much lower than
in naturaolly infdected animals.
protective immune responses = appear to be most effective when
infection occurs before puberty. the majority of infectionbs in
sexually mature animals persist for life although the immune
response does reduce the severity of disease in the uterus and
placenta so that such animals are more likely to carry to term
later in pregnancies.
Vaccines
strain 19 and the McEwen killed 45/20 = two vaccines that are
important in the control of B. abortus infection
Strain 19
= consists of viable culture of a strain that was discovered to
have very little virulence for guniea pigs and cattle but to
possess excellent immunizing properties.
= it has a great stability
= it is a smooth strain that is mildly pathogenic for guniea pigs
= pregnant cattle can be made to abort by inoculating them with
large doses of this
= vaccine organisms can usually be demonstrated without
difficulty in the fetal membranes and the fetus itself
= susceptible cattle, associating with those have aborted as a
result of inoculation with the vaccine strain, do not become
infected
= it is unlikely transmitted from one animal to another; if any
damage is doneby the use of tyhis vaccine, it is limited to the
animal injected.
= it is rarely shed in milk of vaccinated animals
it can cause infection to humans, although it is usually mild with
recovery in a much shorter time than with virulent strains
= it is dangerous to humans and should be handled with caution
= calves are vaccinated 2-10 months of age (vaccination at this
age is advocated to avoid persistent agglutinins that could create
diagnostic problems later in the animal;s life)
= it protects about 65-70 percent of animals for 4 or 5
pregnanciesand is most effective for protection of young
breeding animals when applied in a herd basis.
= animals vaccinated as adults are also protected but develop
persisting agglutinins.
= Latently infected animals or animals in the early stages of
infection do not benefit from vaccination.
Colostral antibodies may interfere with response to strain 19
during the first 5 months of the calf's life.
= vaccination of cows in early pregnancy with large doses
produces probability of uterine infection. this risk is greatly
diminished when the dose is reduced.
= horses with fistulous withers sometimes treated by this
vaccination
McEwen killed 45/20 vaccine
= a rough strain that is inactivated and adjuvanated for use as a
vaccine or antigen in the anamnestic diagnostic test
= it is administered in two doses (6- 12 weeks apart)
= it stimulates complement fixing antibodiesand agglutinins for
rough B. abortus antigens of the IgG1 subclass
= it has been used widely for the control of bovine brucellosis
Diagnosis
bacteriologically and serologically = B. abortus can be
diagnosed by these.
Bacteriological diagnosis
= isolation and recognition of B. abortus are often accomplished
by direct culture on a basal medium such as tryptose agar or
Albimi agar to which serum and selected antibiotics are added.
= cultures are incubated at 37oC in an atmosphere of 10 percent
co2 tension and are examined 2-3 days
=serologic typing of Brucella strains and phage typing with the
Tbilis (Tb)strain of bacteriophage are useful aids to definitive
identification, but their use in restricted to specifically equuipped
reference laboratories
Specimens should be examine by the following:
1. The aborted fetus = direct cultures of specimens of the
stomach contents, intestinal contents, or the lung tissue
2. the placenta = direct stained smears from the outer surface of
the chorion, especially from the margins of the characteristic
thickenings, will usually suffice to make positive diagnosis
without the need for cultural method (the organisms occur free
or enclosed in the epithelial cells). these are visualized using the
modified Koster stain by which appear red against a blue
background.
3. The uterine exudate = after abortion or calving, when the
placenta has been infected in the lochia and can be recognized
by guniea pig inoculation. (organisms seems to disappear and
cannot be found at the uterus until the animal is again pregnant
and reinfection of the organ occurs)
4. milk = udder is infected and detected by the intraperitoneal
injection in milk into guniea pigs or by direct culture
5. abscesses = direct culture of specimens from abscesses of the
testicle and epididymis usually produced pure cultures, and
isolations have been made from hygromas in cattle and from
infected burasae in horses.
Inoculation of guniea pigs = most reliable method of detecting
B. abortus in infected materials. their serums are examined for
the presence of antibodies and specimens from the spleen, liver,
regional lymph nodes, testicles are cultured.
Direct examination of tissues and flourescentantibody = aid to
diagnosis and practicaly useful for examing contaminated
materials (placental membranes, cotyledons, fetal or vaginal
discharges).
serological diagnosis
=the test are applied to serum, whole blood, vaginal mucus,
seminal plasma, whey, or milk.
Serum tube agglutinatio test (SAT) = used in eradication
programs; The test is performed in small test tubes on dilutions
of serum. Complete agglutination at a serum dilution of 1:100
and higher is considered positive. In Great Britain, a titer at least
50 percent agglutination at 1:40 is considered positive
Flase-positive reaction = can be caused by reactions of
antibodies to yersinia enterocolitica 09 and other Gram-negative
organisms. It occur in a significant number of serum sampls
from latently or chronically infected animals. Animals in the
early stage of infection similarly may fail to react.
Buffered antigen plate agglutination tests = replaced standard
SAT because the use of buffered antigen reduces the frequency
of false-positive reactions
Rose bengal plate tests = B. abortus cells stained with rose
bengal and suspended in acidic buffer to inhibit nonspecific
agglutinins. The test can be used as a screen or definitive test.
The frequency of false-negative reactions is less than for the
SAT
Card test = the antigen is a buffered, stained, whole cell
suspension of B. abortus strain. the antibody source is plasma
separated from blood following clumping or erythrocytes by
lectins. the test is useful for the detection of early inection where
IgM is the dominant antibody.
Plate agglutination = test is a heavy suspension of strains of B.
abortus stained withj gentian violet and brilliant green to make
the test easier to read. Serum, whole blood, or whey can may be
used in this test. (using glass slide or plate)
whole-blood method = used for testing range cattle when it is
desirable to hold the animals in chutes until the results are
known
Milk ring test = the antigen is a hematoxylin-stained suspension
of killed B. abortus. It is mixed with fresh milk in a tube in the
proportion of one drop per milliliters of milk. the mixture is then
incubated in a water bath at 37oC for 1 hour.
clumps of agglutinated organisms= a positive test is indicated by
a bluish violet cream layer and a decolorized milk column. The
test is widely used for the examination of bulk milk samples and
so allows herds in an area to be quickly and economically
screened. positive herds are then subjected to blood tests on
individual animals.
complement-fixation test = one of the most accurate tests in
widespread use for the diagnosis of B. abortus infection. It
detects IgM and IgG1 antibodies only. The test does not
differentiate infected animals from animals recently vaccinated
with strain 19 or 45/20. This test designed to test antibody to the
45/20 vaccine is also available. And for that, the test has been
automated in many laboratories thus resulting in greater
precision.
Antiglobulin (coombs) test = measures binding of
nonagglutinating antibodies to B. abortus antigen.
The ravinol and mercapteothenol tests = used to help distinguish
between agglutinin titers caused by chronic infacetion and those
caused by vaccination with strain 19. IgM antibodies are
sensitive to mercaptoethenol.
Enzyme-linked immunosorbent assay (ELISA) = offer promise
of greater sensitivity and specificity than the test described
above.
Allergic skin test = based on purified antigens; is to be a
valuable addition to serology for detecting animals.
Control
1. affected animals are detected and eliminated from the herd
(detection using serological test - milk ring test - agglutination
test - slaughter)
2. resistance of the remaining animals or any replacements is
increased by vaccination
3. General priciples of hygiene are imposed to prevent spread or
reintroduction of infection
systematic calf vaccination = a protective measure in herds that
are only lightly infected
systematic calfhood vaccination with strain 19 or 45/20 = is used
in heavily infected herds
calfhood vaccination = it is maintained in such a herd as long as
there is a real danger of the exposure to the disease from any
source
periodic milk-ring tests and random testing on serum=
surveillance of herd is maintained on bulk milk samples when
they are sent to slaughter
back-tagging = allows infection to be traced back to the her of
origin
Antimicrobial sensitivity
Gentamicin, kanamycin, tetracycline, and rifampicin = B.
abortus is sensitive to these substances
tetracycline + streptomycin = treatment for human brucellosis
*cattle are not usually given antimicrobials for prophylaxis or
therapy of B. abortus infection
Brucella suis
(porcine type of Brucella)
=caused by brucellosis in swine.
=has wider range of host specificty (humans, dogs, rodents,
horses, reindeer, musk, oxen, and wild carnivores)
Five types are recognized = Biotype 1, 2, 3, 4, and 5
Biotype 1, 2 , 3 = have the pig as their natural host but can be
transmitted to other host
Biotype 2 = occurs naturally in horses and relatively avirulent
for humans
Biotype 4 and 5 = reindeer and rodents are the primary natural
hosts (but others can be infected)
Morphology and Staining reaction
B suis is closely resembles B. abortus
Cultural and Biochemical Features
=does not require increased levels of CO2 for growth.
= large amounts of H2S are produced by Biotype 1 but more
produce by other biotypes
= Urea is highly hydrolyzed
Antigen
=surface antigen predominates a smooth cultures of all biotype
except biotypes 4 and 5
Biotype 4 = has both A and M antigens in equal amounts
Biotype 5 = has mostly M antigens on its surface
Epizootiology and Pathogenesis
= it occures to most swine raising areas (not been reported in
Great Britain, Iceland, or Canada)
= Survival in the environment has not been well studied (but
similar to B. abortus)
Feces and urine = the organisms survives at least 6 weeks
Feral swine= is most infected than domesticated swine; can be
also a source of infection for cattle
Biotype 1 = is isolated from tissues of wild swine
wild boars = important reservoirs of B. suis biotype 2
source of infection = contaminated pastures from these hosts can
infect domesticated swine
Reindeer (Rangifer tarandus) = brucellosis is an important
disease; B. suis biotype 4 has been isolated from them; it cause
bursitis; spondylitis, arthritis, and orchitis
Larvae of reindeer warble fly = seen the B. suis organism and
there is a speculation that this insect may act as a vector of this
disease
Wolves of Siberia and sled dogs in Alaska = seen B. suis
because they fed on infected reindeer meat
Soviet Union = brucellosis is of particular interest and
significance because of the large number of their free-living
reindeer
Domestic guard dog = seen to be a carrier of B. suis biotype 2
(source of infection for a herd of domestic swine)
*although feral animals can be a source of B. suis infection for
pigts, the organism is transmitted almost exclusively from pig to
pig (by coitus or by the ingestion of feed contaminated by urine
or genital excretions from infected boars or sows)
* spread is rapid in a susceptible herd
* outbreak of disease followed by intermittent cases as
susceptible newcomers contact chronically infected pigs.
initial infection = B. suis is localized in the regional lymph
nodes. It proliferates and may caiuse extended bacteremia with
generalized infection of the spleen, lymph nodes, joints, udder,
and genitalia
B. suis = a facultative intracellular parasiteand owes a great deal
of its pathogenic properties to its ability to survive in the host's
phagocytic cells.
Erythritol = has a growth-stimulating effect upoin B. suis and is
found in placenta of sows and seminal vesicles of boars
stimulatory action of eythritol + its presence on tissues = lesions
in male and female genital tracts
clinical signs anmd lesions produce = depends on animal's age,
previous exposure, organ or organs involved
Suckling and weanling piglets = they can be affected
adults = can have abortion, metritis, spondylitis (vertebrea in the
lumbar and sacral region are not uncommon), lameness, and
paralysis
posterior paralysis = caused by pressure from the necrotic tissue
on the spinal cord
nodular splenitis = also been associated with brucellosis
Immunity
field and experimental evidence = indicates the swine immunity
to brucellosis is very slight and that after a period of herd
resistance, animals are again susceptible to the disease
Abortion = don't occur after the first exposure but when
reexposed
cell-mediated immunity = important in protecting the host
Diagnosis
cultural methods and agglutination test = positively diagnose
brucellosis in swine
Isolated B. suis = from blood, spleen, uterus, lymph nodes, and
mammary glands of sows and testes and semen of boars
 *methods are the same as B. abortus except that an increase of
the CO2 tension of the culture jar (unnecessary)
Agglutination test = used as an aid in diagnosis but not as
reliable in swine as it is in cattle; it is valuable in determining
whether the infection exists in a herd
Card test and rose bengal test = superior to most other
serological tess for detection of antibody in swine serum
ELISA = it replaced the tube agglutination test
Control
sell all stock to slaughter = simplest way to eradicate the disease
*the premises should be thogoughly cleaned and disinfected
For breeding herd in which blood lines must be preserved = pigs
are raised from the infected unit. They are weaned (8 weeks old)
and tested individually by agglutination test
If negative = they are removed from the infected herd, placed on
a clean ground and raised in isolation
*all pigs are tested periodically (reactors are immediatley
removed)
when of Breeding age = serological negative gilts are bred to
noninfected boars. the original herd is disposed of as soon as the
replacemen unit has grown to sufficient size
Antimicrobial Susceptibility
B. suis is sensitive to = vitro to aminoglycosides, rifampicin, and
tetracyclines
clinical efficacy of antimicrobials = very low
Brucella melitensis
(Bacterium melitensis, caprine type of Brucelle, Micrococcus
melitensis)
=first isolated by Bruce in 1887 from the spleen of aresident of
the Island of Malta who had died of a disease known as Malta
fever or Mediterranean fever
= it is a disease of goats and sheeps
Morphology and Staininbg Reacions
= forms small rods that are so short that they are mistaken to be
a cocci
stains = same as B. aborus
Cultural and Biochemical Features
= does not require supplementary CO2 for growth and either
does not produce H2S (does in trace amounts)
solid and liquid media of B. abortus = suitable for B. melitensis
3 biotype = Biotype 1, 2, 3
Biotype 1 = the most common
Antigens
smooth cultures = have the A and M antigens or both antigens
together.
Epizootiology and Pathogenesis
natural hosts = goats and sheeps
other infected animals = pigs, cattle, camles, and humans, feral
animals including hares and impala
Goats = more susceptible than sheeps
Source = ingestion of feed or water contaminated by uterne and
vaginal discharges from infected does
Milk = contains large numbers of this organisms
Pathogenecity = same as B. abortus in cows
B. melitensis = facultative intracellular parasite that survives and
multiplies within the phagocytic cells of the reticuloendothelial
system
Erythrithol = the growth is stimulated
Extended bacteremia with fever = 5 to 10 weeks or longer is
followed by abortion storm (highly susceptible goat herd or
sheep flock)
Infected does = shows lameness, hygromas, and mastitis
Billy goats = develops orchtis
Kid goats = can remain latently infected until sexually mature
Immunity
Goats and sheep = develop effective protective immune
responses to this organism that eventually clear the infection
Humoral antibody = appears about 2-3 weeks after infection
Infected sheep = develop delayed hypersensitivity to antigens of
B. melitensis
live avirulent Rev 1 and strain 53H38 = vaccines used to prevent
and control B. melitensis
Rev 1 = more effective than 53H38; prevents abortion and
shedding of the organism; animals immunized by this become
serologically negative 6 months later. disadvantage of this is that
it may localize in the placenta of pregnant does and cause
abortion. Localization in the mammary gland and shedding in
the milk also occur
53H38 = stimulates more persistent antibody response then Rev
1
Diagnosis
bacteriological methods = the same as those used to detect
brucellosis
phage sensitivity, agglutination with monospecific antiserum,
guniea pig inoculation = isolations may be identified
complement fixation, agar-gel immunodiffusion, and rose bengal
plate test = serological tests tat are available and give
comparable results with the classic agglutination test
Whey test = can be performed also
Milk-ring test = not reliable on sheep milk
Intrapalpebral allergic test = more sensitive but slightly less
specific than serological assays for detection of infection in
sheep flocks
Antimicrobial Susceptibility
Aminoglycosides, rifampicin, and tetracyclines = sesitive to B.
melitensis
Efficacy of antimicrobials in therapy = has not been reported
Brucella neotomae
= first isolated by Stroenner and Lockman from a dessert wood
rat (Neotoma lepida) trapped alive in the Great Salt Dessert of
Utah
= it has not been recovered from any other naturally infected
host
wood rat = well tolerated and upon experimental infection it
persist for at least a year without producing significant lesions
Brucella ovis
=causes epididymitis in rams
= early isolation in New Zealand and Australia
= produces late abortions in females and lowers flock fertility

Morphology and Staining Characteristics
=acid fast under certain conditions of staining
korster staining method = it stains blue, not red
cultural and biochemical features
=requires supplementary CO2 for growth
= does not produce H2S
=most strains are urease negative
= does not metabolized erythritol or other carbohydrates
colonies = appear to be smooth, they are always in the rough
phase on primary isolation
= there are no biotypes
= the only species to show less than 100 percent DNA homology
with otehr species in the genus
Antigens
= it has an R surface antigen that cross reacts with roughj strains
= does not react with A or M antigens
= shares a large number of protein antigen with other brucella
sp.
Epizootiology and Pathogenesis
sheep = only natural hosts of B. ovis
British breeds = more susceptible than Merinos
rams = clinically affected much more frequently than ewes
Transmission = venereal, and rams become infected by mating
with ewes previously covered by other infected rams
ewes = developed placental infections with subsequent abortion
Pasture = B. ovis can survive for a month or two
Route of natural infection = ingestion; rams can be be
experimentally infected by the oral route as can ewes in the early
stages of pregnancy; rams as young as 4 months old can be come
infected
clinical signs = transient, inaapparent bacteremia with
subsequent localization of the organism in the epididymis of the
male. (tail of the epididymis is affected often unilaterally). the
ram may show no clinical signs at this time but shed the
organism in its semen for a prolonged period .
epididymis = a permatocele forms and ruptures, and finally
spermatic granulomas form with later testicular atrophy
Rams= have lowered fertility
Type 3 hypersensitibity responses = plays a role in the
pathogenesis of the epididymis because lesions in
experimentally challanged rams are more severe in animals
previously vaccinates than in nonvaccinated controls.
Ewes - less susceptible to infection by B. ovis but lesions occur.
They vary from a superficial purulent exudate on an intact
chorioallantoic membrane to advanced fibrosis and necrosis of
this membrane
Lambs = organism is pathogenic to this in the utero, but the fetus
may survive in the presence of infection
Placentitis = interferes with fetal nutrition
Immunity
cell-mediated = immunological basis of the protective immune
response
*many rams that become infected do not develop epididymitis
humoral antibodies = detectable in about 3-4 weeks after
infection, and delayed cutaneous hypersensitivity to B. ovis
antigens develops later
vaccines = consists of saline oil adjuvant bacterins of
formalinized B. ovis or live B. melitensis Rev 1
B. ovis 7-26 = a live vaccine was shown to be effective in 6-10
weeks old lambs (it was not transmitted to unvaccinated sheep)
Vaccination of ewes = does not influence spread of infection in a
flock
Diagnosis
=palpation of the testicles, culture of the semen, and
demonstration of antibodies in the serum
Palpation = is of limited value since many advanced cases show
no palpable lesions of the testes (enlarged epididymis with
testicular atrophy is valuable in diagnosis)
culture of the semen = important adjunct to diagnosis and has
been facilitated by the use of selective media modieifed for the
growth of the organism
flourescent antibody test = use to detect the organism in a semen
smear
complement-fixation, ELISA, immunodiffusion, and
hemagglutination test = used for the detection of the antibody in
the serum
ELISA = less accurate and sensitive as the complement-fixation
test nad is easier to perform
complement-fixation test = rams are five times more likely than
ewes to be positive
Control
= depends on preventing spread of infection between rams
Infected older rams = must kept separate from young rams and
the latter are used only on ewes known to be free of infection
eradication of the disease = reacts in the complement-fixation
and ELISA
Brucella canis
(canine type of Brucella)
first isolation = in 1966 from outbreaks of abortion and whelping
failures among dogs in the US and Great Britain
1968 = it was characterized and named Brucella canis
= highly adopted to domestic dogs
=not readily transmitted to other animals, although it will infect
humans.
Morphology and Staining reactions
=a small rod-shaped coccobacillus similar to other Brucella sp.
Stains = similar to other Brucella sp.
Cultural and Biochemical Features
=produces only rough or mucoid colonies
= smooth phase has not been observed
= it is inhibited by 10 percent CO2
Broth =growth becomes quite mucoid and a ropy, viscous
sediment is formed
=does not utilize erythritol as a preferred nutrient
=produces large amounts of urease
= does not form H2S.
Antigens
= it carries R but not A or M surface antigens
= s species-specific antigen is associated with the mucoid
antigen shed by the organism
= cross-reactivity with Actinobacillus equuli, Pseudomonas
aeruginosa, and P. multicoda also occur
= it has been found in breeding kennels, pet and stray dogs, and
in lkaboratory colonies of beagles
= infection appears to be more prevalent in stray dogs than in
pets
= 2-3 years age group had a higher prevalence of seropositivity
than did older dogs
Transmission = occurs proncipally at the time of abortion, when
mayn bacteria are shed in the persistent vaginal discharge. it
continues 4- 6 weeks after an abortion
Male dogs = harbor the organism in their genital tracts, from it
shed intermittently (transmitted by coitus to the female)
=after initial infection, bacteremia follows for 2 years.
= infected dogs do not show no clinical signs but some do
exhibit clinical signs have generalized lymphadenitis, splenitis,
and embryonic deaths and abortions at approximatelt 50 days of
gestation.
Infected males = have epididymitis, scrotal dermatitis, and
testicular atrophy.
Sperm agglutination in the semen, phagocytosis of spermatozoa,
delayed skin hypersensitivity to testicular antigens = isioimmune
responses to sperm antigens plays a role in the pathogenesis of
B. canis infections and male fertility
*there is evidence that damage to epithelial cells of the testes,
epididymis, and prostate caused by B. canis (triggers production
of sperm antibody)
Immunity
cell-mediated = protective immune response of B. canis
Humoral antibodies = appear at between 2-7 weeks nad are
present in highest concentration in bacteremic animals
Titers = diminish with resolution of the bacteremia
*there is no successful vaccines against B. Canis infection in
dogs
B. ovis vaccines = can stimulate some resistance to B. Canis, but
this resistance is much inferior to that observed dogs following
recovery from B. canis infection
Diagnosis
*diagnosis is easy in animals that show clinical signs of abortion
and infertility (females) or epididymis (males)
Clinical signs, direct cultre of blood, lymph nodes, or bone
marrow; and serological test = are used in diagnosis
mercaptoethanol, tube agglutination, complement-fixation,
counter immunoelectrophoresis, agar-gel immunodiffusion, and
a rapid slide agglutination test = are not completely standardized
and are subject to occassional but important "false-positives" and
hard to interpret "low titers" and to broad heterotypic reactivity
shown by B. canis antigens
Agar-gel immunodiffusion test = based on internal antigens of
B. canis is the most sensitive and accurate serologic means of
detecting infection.
Isolation of B. Canis in infected dogs = most certain diagnostic
method
Antimicrobial Susceptibility
Vitro to aminoglycosides, rifampicin, and tetracycline = B. canis
is sensitive to
*B. canis infection is unlikely to be successful
aminocycline (25mg/lb twice daily) + streptomycin (25 mg/lb) =
given by intramascular injection for 7 days is effective in curing
only 3 out of 11 experimentally infected dogs.
Brucellosis (Undulant Fever) of Humans
= can be caused by B. abortus, B. canis, B. melitensis, and b suis
(all biotypes)
=species involved is in great part determined by opportunity for
exposure
B. suis infection = common in swine belt (north central of US)
B. abortus = southeastern US is found
B. melitensis = coomon in Mexico and in localized areas of the
southwestern part of US
=this species is also found with some frequenxy in the
midwestern states (where it is contracted from swine)
Caribou (North American reindeer Rangifer tarandus) =
brucellosis in humans have been derived in Alaska infected with
B. suis biotype 4
B. canis = many peopele were found to be infected with this
species ten years after it was isolated. Half were laboratory
personnel who were working with the organism, and most of the
owners of infected dogs were infected
Cases of B. abortus and B. suis = usually seen in veterinarians,
farmers, slaughterhouse personnel, and others connected to cattle
and swine
=men are more susceptible than woman (2/3 of the cases were
men that was infected partly B. abortus and partly B. suis
infection)
direct contact = is more hazardous than drinking infected milk
(infections in men are more common even in areas where the
predominant type is B. abortus)
20 and 45 years = age group that can have brucellosis
infants = who drinks milk is seldom become infected although
the disease has been diagnosed in children as young as 4 years
old
B. suis = more virulent for humans then B. abortus; most often
contracted by exposure to blood or tissue fluids from infected
swine in the slaughterhouse
Brucellosis as an Occupational Hazard for Veterinarians
brucellosis = poses a hazard to veterinariansin rural practice
because they often come in contact with infected secretions
Blood test surveys = indicates that a comparatively large
percentage of veterinarians reacted positively, although may had
no clinical history of the disease
infection in veterinary colleges = was not uncommon
*the veterinarians who showed no evidence of antibodies to
brucella infection were either recent graduates or involved in
commerce, laboratory work, or teaching.
practicing veterinarian = is exposed to more viable Brucella
organisms than other people in rural communities. during the
common procedure of removing a retained placenta from a cow,
the vet. comes in intimate contact with the uterine discharges
rich in organisms that can enter his or her body through the
conjunctiva or the intact skin, or by inhalation.
wounds on the hands = make ideal sites of entry
Accidental inoculation with B. abortus strain 1 9 = major source
of infection in vet in vaccinating cattle. Veterinarians may jab
the needle into his thigh or scratch thumb.
strain19 = it usually does not cause chronic infections in humans
but can cause severe effects in those previously infected with B.
abortus
Diagnosis
*the acute disease is frequently overlooked or misdiagnosed.
*patients frequently decide they have influenze or chills and then
recover
acute stage = the disease may be misdiagnosed because of its
sporadic in nature and its vague signs
diagnosis = based on the patient's history, clinical signs,
isolation of the organism, and serological tests.
severe phase of the disease = individual is acutely ill, suffers
from prostration and weakness, develops daily fever (afternoon
and evening), suffers chills and night sweats during which fever
disappears only to have the cycle recur on following days. the
acute symptoms usually lessen after a few days, but , following
an interval of varying length during which the patient feels
better, another period of acute symptoms may appear. There may
be several remissions
intermittent fever = responsible for the name undulant fever
Infections with B. melitensis and B. suis = are usually more
severe than those with B. abortus, but this is not always the case.
*the mortality is low, but recovery from infections often is very
slow.
*many persons never fully recover from the effects of this
disease.
chronic cases = this form is more difficult to diagnose. usually
the patients suffers from great debility, weakness, a low-grade
remittent fever, and joint pains; there may be sweating, lassitude
and malaise, gastritis, abdominal pain, skin rashes, headache,
irritability, depression, insomnia, arthritis, and backache.
*patients may be lebeled neurotic (complains are gard to
substantiate by laboratory test)
Melitococcic spondylitis = an adteoarticular complication of
brucellosis of humans in Italy
septic arthritis of the hip= came from B. suis
Brucella organisms = implicated cases of osteomyelitis and
diseases in the nervous system
blood culture = when positive, are diagnostic
isolation of the organism = from the blood is usually dirricuylt
and often possible, particularly when the offending organism is
B. abortus
*Greater success is achieved in acute rather than in chronic cases
Standard agglutination test, antiglobulin test, and the
complement-fixation test = three serological test used in aid to
diagnosis
Agglutination test = it has been used for years, but it must be
interpreted with great caution because it may produce aberramt
results, such as occassional prozones, and "nonagglutination"
antibodies may be present, giving a negative test even in persons
who have had the disease for a long time; useful particularyly in
recently acquired acute cases of brucelllosis
Antiglobulin test (Coomb's test) and complement fixation test =
are supplementary and are value in the diagnosis of long-
stranding brucellosis
Treatment
Tetracyclines = are recommende by FAO/WHO Expert
Committe on Brucellosis be given for 21 days
Tetracycline + streptomycin = must be given to severe infection
and for all B. suis infections
Chloramphenicol = is of value but urged caution in its prolonged
use.
*although these antibiotics often produce a good
initial clinical response, a large percentage of patients
suffer recurrence of the disease.