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Canine Brucellosis antigen test kit 10 kits a box

Canine Brucellosis, caused by Brucella canis, is a significant reproductive disease of dogs. It is caused by
an intracellular bacterium and often found in breeding kennels throughout the United States.
Brucella canis is a zoonotic organism that can infect humans and is currently considered to be underreported in human
medicine.
The true prevalence of the disease in dogs is not well understood.
The prevalence rates mentioned in currently available review articles and information sheets reference results obtained in the 1980’s and 1980’s with reported rates of infection of1 – 8%.
Within the past few years it has become apparent that
the incidence of B. canis infection within the dog breeding industry has been on the rise.
Brucellosis may have also become more recognizable due to increased awareness coupled with improved
diagnostic testing capabilities by Genprice

There is much confusion about the disease among kennel operators and veterinarians alike. Brucellosis,
while historically thought of as a disease that causes abortions, has many clinical signs that are often
misinterpreted. These include but are not limited to early abortions, testicular swelling, uveitis and
spinal arthritis. Misunderstanding about the tests available for Brucellosis, often leads to the disease in
kennels being misdiagnosed or under diagnosed. Routine blood work and urinalysis are often normal.
It is possible for infected females to raise infected puppies that can enter consumer markets.

A 2011 survey of State Public Health Veterinarians reported that B. canis infection is a reportable disease in at
least 28 states. 1 Because the disease is reportable in many states there is a small but important
“underground” that tries to avert the reporting and thus serves as a continuum for the disease.
Additionally, it is important to emphasize that in dogs it is not a curable disease, which means that
carrier animals MUST be removed from the breeding population in a kennel situation. Attempts at
treatment have been very disappointing with relapses commonly occurring. Attempted treatment can
mask diagnostic testing and has been shown to be yet another important contributing factor in the
spread of the disease.

Pathogenesis and Clinical Signs

Natural transmission of canine brucellosis can occur by several routes. B. canis organisms are shed in
the highest numbers in aborted material and vaginal discharge. Infected females transmit canine
brucellosis during estrus, at breeding, or after abortion through oronasal contact of vaginal discharges
and aborted materials. Shedding of B. canis may occur for up to six weeks after an abortion. Semen,
seminal fluid and urine from infected males have also been documented as sources of infection. The
rate of isolation from the semen of B. canis is usually high for the first six to eight weeks after infection
but males can have intermittent shedding of the organism in low numbers for up to two years. Both
males and females may shed the organism in the urine for at least three months after becoming
infected. The organism can also be present in blood, milk, saliva, nasal and ocular secretions, and in the
feces. Brucellosis has also been documented to be spread via fomites during vaginoscopy, artificial
insemination, blood transfusion, or by the use of contaminated syringes or other medical and husbandry
equipment (do not share kennels, crates or other equipment). Hands and clothing can also act as
fomites and increase the spread of the organism within a kennel (do not visit other kennels without
washing clothes, disinfecting shoes and washing your hands). The use of high pressure spraying and
cleaning equipment and improper wash-down techniques within kennels have also been shown to
facilitate the spread of the disease. Puppies can become infected in utero, during birth, while nursing,
by contact with contaminated surfaces, or by fomite spread.

Canine brucellosis is considered a lifelong infection in dogs. Clinical signs of canine brucellosis are
extensive and variable and depend on the sex and age of the dog affected. The disease often exhibits no
clinical signs that are obvious to the owner or veterinarian, or it can present as an abortion storm
affecting many dogs. B. canis is a major cause of infertility and abortion in breeding dogs and therefore
should be one of the first diseases to be ruled out when investigating reproductive complaints.
Several months after infection, there will be evidence of conception failures and infertility, such as lateterm abortion with prolonged vaginal discharge, stillbirths, decreased litter size, and decreased puppy
survivability. As the disease progresses adult dogs may develop epididymitis, orchitis, testicular swelling
or atrophy, sperm abnormalities, uveitis, meningioencephalitis, spinal arthritis, weight loss, poor hair
coat, listlessness, swollen lymph nodes, and behavioral changes. Some chronically infected dogs may be
totally asymptomatic and still be a source of infection for other animals and humans. The clinical signs of
infection can mimic many other diseases, and for that reason, Canine Brucellosis has been described as
“The Great Imposter”.

In the female the disease is usually thought of as a cause of abortions in affected breeding dogs. Most
abortions occur between 45 and 55 days of gestation, and the aborted fetuses appear autolyzed. Early
embryonic death may lead to resorption and result in perceived conception failure. Endometritis is
usually seen in cases of uterine infections. It is also common to observe a prolonged vaginal discharge
either following an abortion or during proestrus. The female may also have puppies that are born dead,
born alive and soon die, or in some cases puppies that live to become adults.

In the male B. canis usually infects the prostate, testicles and epididymis and clinical signs are
associated with these areas. Enlargement of one or both of the testicles may be seen. In long standing
cases atrophy of the testicles may occur. Enlargement and inflammation of the epididymis
(epididymitis) is a commonly observed clinical sign. Scrotal dermatitis and swelling is usually a source of
intense licking by the affected male. Infertility may be seen, but these infected males may also breed
successfully.

The organism’s effect on spermatozoa is well described by Greene and Carmichael. Semen abnormalities can be identified starting five weeks post infection (PI) and become pronounced by eight
weeks. These early abnormalities include immature sperm, deformed acrosomes, swollen midpieces,
and retained protoplasmic droplets. By 15 weeks PI bent tails, detached heads, and head-to-head
agglutinations are seen. By 20 weeks PI 90% of sperm are abnormal. Aspermia without inflammatory
cells may develop and is associated with testicular atrophy.

Generalized signs may be seen in dogs of any sex or age and include lethargy, lymphadenitis, ocular
disease and vertebral pain. Loss of libido and unwillingness to breed are common signs seen in breeding
stock. B. canis should be considered whenever reproductive problems occur, whether acute or chronic.

Zoonotic Information concerning Brucella canis

To decrease the possibility of human exposure to B. canis veterinarians should encourage their dog
breeding clients to wear single-use protective examination gloves during assistance with whelping
including the handling of newborn puppies, placentas, fetal membranes, or exposure to urine or vaginal
secretions. Extreme caution should be taken when handling any aborted materials including dead or
partially developed puppies, their fetal membranes and placentas. Protective gloves should also be used
during assistance with any natural or artificial breeding. Appropriate use of personal protective
equipment beyond just wearing gloves (respiratory and ocular protection) may be required to prevent
human infection during cleaning and disinfection and animal handling in brucellosis positive kennels
during quarantine and isolation situations. Caution should be taken when collecting and handling blood,
serum, fluids or tissues for laboratory analysis by the veterinarian, paraprofessional staff, the client, and
laboratory personnel if brucellosis is suspected.

Veterinarians should discuss with their dog breeding clients the potential for liability and damaged
reputation surrounding zoonotic cases of B. canis entering consumer markets through the sale of
infected puppies or adult dogs. These puppies or adult dogs commonly come into contact with children,
older adults and other immunocompromised individuals. A recent example is a 2012 New York City case
involving a 3 year old girl which is the first documented case of transmission of B. canis from a puppy to
a child in the United States. This human case of B. canis infection resulted in a multistate, multi-agency
investigation to determine the source of the child’s infection, origin of the puppy, location of the
puppy’s littermate in another state (PA), additional human exposures, and an investigation of the
Midwest breeding facility (personal communication with the CDC and the New York City Department of
Health and Mental Hygiene).

The symptoms of B. canis infections in humans are generally similar to those of brucellosis caused by the
other Brucella species (e.g., B. abortus and B. melitensis). They are frequently non-specific, and may
include one or more of the following: fever (often periodic and nocturnal), fatigue, headache, weakness,
malaise, chills, sweats, weight loss, hepatomegaly, splenomegaly, and lymphadenopathy.
A thorough review of the current understanding of B. canis infection and the diagnostic challenges in
humans can be found in the 2012 National Association of State Public Health Veterinarians (NASPHV)
document “Public Health Implications of B. canis Infections in Humans” 1 from which the following two
quotes have been extracted:

“Although there are multiple statements in the literature that B. canis infections tend to cause a
milder illness compared to other Brucella spp., serious manifestations have been described. These
include septic arthritis, aortic valve vegetations, calvarial osteomyelitis, epidural abscess, pleural
effusion, oral lesions, lower extremity aneurysms, and culture negative endocarditis. There are at
least two reports describing B. canis infection in HIV-infected patients. The disease in both patients
was well-controlled with regard to viral load and CD4 counts, and each had typical clinical
presentations of brucellosis with good responses to treatment. “

“Although the low numbers of known human cases imply that the impact of B. canis on human
health is minimal, it seems likely that a lack of clinical suspicion of the infection, its nonspecific clinical
presentation, the non-availability of approved serologic tests, and the organism’s fastidiousness in
culture all result in the underdiagnosis, and subsequently the underreporting, of this infection”.

Population Medicine and Biosecurity Considerations

Ideal control measures for canine brucellosis in breeding kennels centers around sound population
medicine and biosecurity practices. Gaining negative Brucellosis status involves testing and removal of
infected animals, sourcing new additions to the breeding population from proven negative kennels, and
isolation and repeat testing of new additions to prove them brucellosis negative. Practicing strict
biosecurity, continual disease surveillance with proper breeding management and maintaining strict
environmental controls (cleaning, disinfection, proper temperature and humidity) are required to
maintain Brucellosis free status. Particular care should be taken to properly clean and disinfect whelping
sites and nurseries on a daily basis.

Once a breeding dog population is proven to be free of brucellosis the best way to keep brucellosis out
of the kennel is to isolate and test all incoming dogs, proving them negative prior to placing them into
the general kennel population. This is best accomplished by isolating newly purchased dogs in a
separate building or facility, away from the general population for a minimum of eight weeks. All
incoming dogs should be tested for B. canis on arrival and again at eight weeks. Only after these two
negative screening tests have been done on all dogs in the isolation facility can they be safely moved
into the established kennel. If during this eight week isolation period any of the dogs in isolation are
diagnosed with brucellosis they should be eliminated from the premises immediately. The eight week
isolation period is then restarted for the remaining dogs in isolation. This isolation and testing approach
has proven to be the safest way to introduce new dogs into an established breeding population without
the fear of introducing brucellosis or other infectious organisms.
Ideally, breeding dogs should never leave the breeding kennel facility other than to visit a veterinarian
for necessary treatment (e.g., C-section, serious injury or illness). It is best to maintain a closed
population of breeding dogs in the kennel and not send females out to be bred. A female sent out for
breeding purposes is a risk and any dog leaving the breeding kennel facility for any reason other than a
C-Section should be tested 90 days after its return to confirm its negative status. It is best to isolate that
dog from the rest of the kennel population upon its return, but doing so may not be practical because it would require isolation during gestation and whelping, which could be problematic in some kennel
situations.

An alternative approach for bringing in new genetics would be to use artificial insemination (AI) for
breeding females, with semen obtained from outside stud dogs that have been proven to be negative
for B. canis (two negative tests at least eight weeks apart prior to collection of the semen). If male dogs
are to be used for outside breeding, the safest approach would be to only offer this service through the
use of AI, with the semen collected on the premises and then shipped to the outside kennel without the
female ever coming to the facility.

Visitors (including the breeding client themselves) should not have visited any other breeding facilities
that day, should be wearing clean clothing, should disinfect their shoes, should wear disposable
protective shoe covers, and should adequately wash their hands. Ideally visitors should not touch or
handle any dogs or equipment. Some accomplish access without direct contact using video viewing of
dam and sire as well as video of the litter through the nursing period. This eliminates most direct
disease transfer risk until the puppy is moved to the new home.

Cleaning and Disinfection

B. canis is relatively short-lived outside the body and is readily inactivated by common disinfectants as
well as by sunlight. B. canis is stable in the environment in the presence of organic debris for up to two
months (proper cleaning and disinfecting is therefore a must). B. canis can withstand drying in the
presence of organic debris, can withstand freezing, and can survive in water, dust and soil. The
combination of organic debris, high humidity, low temperatures and little or no sunlight favors survival
of the organism (winter conditions in most dog kennels).

An important and often overlooked part of kennel management is the proper cleaning and disinfection
of a kennel. When dealing with Brucellosis or other diseases cleaning and disinfection serves to limit the
spread of the disease and is a critical component in disease prevention. Proper cleaning and disinfection
takes time and must be done correctly for a kennel to be considered truly disinfected. It is important to
remember that a clean kennel is not always a disinfected kennel. The consulting veterinarian must be
certain that their clients thoroughly understand the entire cleaning and disinfection process including
proper dilution and storage of the cleaners and disinfectants, and the fact that required contact times
and appropriate rinsing are absolutely necessary. The required frequency (daily vs. weekly) for cleaning
and disinfection should be thoroughly discussed with the client. The facility should always be cleaned
and disinfected in order of animal susceptibility to disease starting first with the kennel areas housing
the most susceptible animals (puppies and nursing bitches), followed by healthy adults and lastly areas
housing any unhealthy or isolated animals. Clients should be educated on the specific biosecurity
procedures that would be required during isolation or quarantine situations.

If there is a “good” aspect of B. canis, it is that the organism is very sensitive to most disinfectants when
they are used properly. The use of a biodegradable enzyme based kennel degreaser is a must before
disinfection. A properly used degreaser will effectively remove the biofilm and other organic debris (e.g.,
residues from feces, urine, vaginal or preputial discharges, oil from the skin and haircoat, other body fluids) that build up in all kennels and can “hide” the offending pathogen(s).

The degreaser can be applied directly with a spray bottle or conveniently by the use of a hose end foamer. The foamer allows
for ease of application and increases the surface area covered. It can also help to ensure that all surfaces
are covered. The use of degreasers that leave a soapy residual film that can contribute to the biofilm is
discouraged. It is important to rinse all surfaces well after using any degreaser. Once a kennel has
undergone degreasing the final step in the cleaning and disinfection process requires the proper
application of an appropriate disinfectant to kill the offending pathogens. A properly functioning hose
end foamer supplying the correct concentration of disinfectant is an excellent way to apply the
disinfectant to all surfaces that the dogs will come in contact with. It is important to remember that all
disinfectants have a minimal ten minute contact time with the surfaces to be effective (follow product
labels and instructions supplied with disinfectant). Most disinfectants require adequate rinsing with
clean water after the appropriate contact time has occurred. After following these procedures we can
be assured that we have reduced the biofilm and disinfected the kennel.

Diagnosis of Brucella canis

B. canis monitoring may be achieved with a variety of tests which have been well described by Hollett 5
and by Greene and Carmichael. 6 See the test comparison chart below for specific information on
available tests. The chart was compiled from various sources including peer reviewed publications 5,6,
available product inserts 7
, and information obtained from reputable veterinary diagnostic laboratories.
Because of the zoonotic potential of Brucella, caution should be exercised when collecting and handling
blood, serum, fluid or tissues.

Comparison of Diagnostic Procedures for B. canis
Antibody Detection Methods

Test Sample
Earliest
Detection
(weeks Pl)
Advantages Disadvantages
Rapid Slide Agglutination
Test
(RSAT)
Serum 1- 4 weeks*
3-4 weeks**
Quick, high sensitivity, few false
negatives. Good Screening Test.
False positives possible, must confirm
by other tests.
Mercaptoethanol (ME)
rapid slide agglutination
test (ME-RSAT)
Serum 3 – 4 weeks*** Quick, high sensitivity, few false
negatives. Increased specificity over
RSAT.
False positives possible, must confirm
by other tests.
Tube agglutination test
(TAT)
Serum
2-4 weeks**
3-6 weeks***
Semiquantitative titer. Good
Screening Test.
False positives possible, must confirm
by other tests.
.
ME-TAT
Serum
2-4 weeks**
5-8 weeks***
Semiquantitative titer. Increased
specificity over TAT Longer testing time (2 day test).
Agar-gel immunodiffusion
(AGID) cell wall (somatic)
antigen
Serum
8-12 weeks**
5 – 10 weeks***
Positive earlier than CPAg
Very sensitive test.
Procedure and interpretation complex,
nonspecific reactions, poor availability.
Internal cytoplasmic
protein antigen (CPAg)-
AGID
Serum 8 – 12 weeks***
Highly specific confirmatory test
utilizing highly purified cytoplasmic
protein devoid of contamination
with LPS.
Maternal antibodies prevent
seroconversion in puppies; so not useful
until 6 months post weaning. Complex
procedure.
Indirect Fluorescent
antibody
Serum
Unknown
Available and convenient for
diagnostic labs ***
Good Screening Test.
May be less sensitive than ME-TAT as
screening test ***
False Positives Possible.
ELISA
Serum 30 days***
Good results with mutant (M-) B
canis for cell wall extracts, or B
abortus for CPAg ***
Antigen purity and preparation critical***

* D-Tec CB Canine Brucellosis Antibody Test Kit directions insert; Zoetis Animal Health, Florham Park NJ, USA
** Hollett, R.B., 2006; Canine Brucellosis: Outbreaks and Compliance; Theriogenology 66: 575-587
*** Greene, Craig E. and Leland E. Carmichael, 2012: Chapter 38: Canine Brucellosis; in Infectious Diseases of the Dog and Cat, Fourth
Edition, Craig E. Greene (editor).

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Qualitative Detection of Dirofilaria immitis Specific Antigens