wildlife and zoo assay data sheet
mycobacteriosis in amphibians
Panel code: P0030 -
Ultrasensitive detection of
Mycobacterium species commonly infecting amphibians, by real time PCR.
Panel P0030 includes:
code B0070: Mycobacterium xenopi
Test code B0071: Mycobacterium chelonae
code B0072: Mycobacterium liflandii / ulcerans / marinum /
shottii / pseudoshottii
not detect other mycobacterial species.
included in P0031 -
amphibian screening panel
genus Mycobacterium contains more than 70 species, which
can be divided into two groups based upon their growth rate in
culture. Slow-growing species require more than 7 days to be
visible on culture, whereas the rapid-growing species require
less than 7 days. In general, many slow-growing species are
pathogenic in humans, whereas rapid-growing species are usually
in amphibians, especially frogs, is caused by acid-fast bacilli
of the genus Mycobacterium.
Many mycobacterial species found in the environment can cause
infection in amphibians, including
M. chelonae, M. ulcerans,
M. liflandii and
M. xenopi (Mve-Obiang et al., 2005, Trott et al., 2004). All these
mycobacerial species are ubiquitous in nature and can be found
in water. They are resistant to normal water treatment.
is a serious threat to the health of a frog colony, although
tuberculosis usually only results in frogs which are already
ill, are stressed or have weakened immune systems. Infected
frogs will usually develop cutaneous and visceral milliary
granulomas (white millet-like bumps) and ulcerative lesions on
the skin. Any of these mycobacteria can also gain entry to a
frog’s body through the skin wounds and lesions. Once through
the skin, these mycobacteria can also affect internal organs.
is a slow-growing, waterborne, scotochromogenic species of
Mycobacterium. It was first isolated from skin lesions of
the frog, Xenopus laevis (Schwabacher, et al., 1959). The
organism is very resistant to most disinfectants used to clean
water systems and has been isolated in samples collected from
water systems in homes and hospitals.
Mycobacterium chelonae is a fast-growing, nontuberculous mycobacterial species. It
is a normal commensal in water and soil that can cause disease
in humans, fish and amphibians. Recently, this mycobacterium was
isolated from captive South African clawed frogs (Xenopus laevis)
showing chronic weight loss and nonhealing ulcerative skin
lesions (Green et al., 2000), and also from dolphin showing
pyogranulomas in blubber, as well as marked acute multifocal
necrosuppurative pneumonia and lymphadenitis (WŁnschmann et al.,
can cause a mortality rate of 90% in
infected immunocompromised human patients such as AIDS or cancer
patients. In humans, infection is usually via the pulmonary
Mycobacterium liflandii / M. ulcerans / M. marinum / M. shottii / M. pseudoshottii infection of Xenopus tropicalis
can also result in coelomitis (infection of the coelom/abdomen)
and subsequent general or local edema (bloating). Cutaneous
lesions and bloating can occur independently. Once these signs
develop, the disease is consistently fatal and affected frogs
are usually euthanized.
Diagnosis of infections caused by mycobacteria, especially
nontuberculous mycobacteria, remains a difficult task both in
microbiology and pathology. Culture identification followed by
biochemical analysis is often slow and may not differentiate
pathogenic from nonpathogenic mycobacteria species due to their
close similarity. False positives occur in this way in both
staining and culture, particularly in environmental specimens.
Molecular detection by PCR, on the other hand, is a rapid,
sensitive and specific technique for sensitive mycobacterial
detection and differentiation.
his panel detects and
differentiates M. xenopi
and M. chelonae; it detects but does not differentiate the group
M. liflandii / M. ulcerans / M.
marinum / M. shottii / M. pseudoshottii, as these last
five species are genetically
Help confirm the disease causing agent
Shorten the time required to confirm a clinical
Help ensure that animal facilities or amphibian populations
are free of these mycobacteria
Early prevention of spread of these mycobacteria in a
facility or geographic area
Minimize human exposure to these mycobacteria
Safety monitoring of biological products that derive
from susceptible animals
Schwabacher H. (1959) A strain of Mycobacterium isolated from
skin lesions of a cold blooded animal,
Xenopus laevus, and
its relation to atypical acid-fast bacilli occurring in man. J.
Mve-Obiang, A., Lee, R.E., Umstot, E.S.,
Trott, K.A., Grammer, T.C.,
Ranger, B., Grainger, R., Mahrous, E.A. and Small, P.L.C. (2005)
A newly discovered mycobacterial pathogen isolated from lethal
infections in laboratory colonies of Xenopus species produces a
novel form of the M. ulcerans macrolide toxin, mycolactone.
Infect. and Immun. 73: 3307-3312.
Stacy, B.A., Lifland, B.D., Diggs, H.E., Harland, R.M., Khokha,
M.K., Grammer, T.C. and Parker, J.M. (2004) Characterization of
a Mycobacterium ulcerans-like infection in a colony of African
Tropical Clawed Frogs (Xenopus tropicalis) Comp. Med. 54:
Green, S. L.,
Lifland, B. D., Bouley, D. M., Brown, B. A., Wallace, R. J., Jr.
and Ferrell, J. E., Jr. (2000) Disease attributed to
in South African clawed frogs (Xenopus
laevis). Comp. Med. 50: 675-679.
WŁnschmann, A., Armien,
A., Beth Harris, N., Brown-Elliott, B.A., Wallace R.J., Jr.,
Rasmussen, J., Willette, M. and Wolf, T. (2008) Disseminated
Panniculitis in a Bottlenose Dolphin (Tursiops
truncatus) due to
Infection. J. Zoo Wildlife Med. 39: 412-420.
or environmental surface swab, or 0.2 ml fresh tissue, or
0.2 ml fixed tissue, or biofilm swab from filter media or tank
Tissue or swab samples may be placed in 70% ethanol if desired;
if so, ethanol volume should be minimized - only use enough
ethanol to cover the head of the swab or the piece of tissue.
if advice is needed to decide which sample type is appropriate
for the specific diagnostic application. For specimen
types other than those listed here, please call to confirm
specimen acceptability and shipping instructions.
specimen types, if there will be a delay in shipping, or during
very warm weather, refrigerate specimens until shipped and ship
with a cold pack unless more stringent shipping requirements are
specified. Frozen specimens should be shipped so as to remain
frozen in transit. See shipping
instructions for more information.
2 business days
Qualitative real time PCR