environmental, wildlife and zoo assay data sheet
- Ultrasensitive qualitative screen for
real time polymerase chain reaction. This screen
detects but does not
Test P0008 is
- waterborne pathogens screening panel
and in P0047 - ruminant fecal
- Ultrasensitive qualitative detection of
by real time polymerase chain reaction.
the 13+ species in the
Cryptosporidium genus have been confirmed as
causative agents of human disease.
Cryptosporidium is a
parasitic protozoan that is transmitted by multiple routes; the
animal host range is diverse. The following
species are currently accepted, on the basis of host
specificity, pathogenesis, morphology and genotyping:
C. parvum, C. wrairi, C. felis, C. canis, C. andersoni,
C. muris and C.
birds: C. baileyi, C.
reptiles: C. serpentis
fish: C. molnari
analyses have been largely based on sequencing of the small
subunit rRNA gene (18S rRNA), the hsp 70 gene, or other
housekeeping or structural genes. These analyses reveal that the
species interact in complex ways with hosts. For example, the
specific host of C. felis
is cats, but this species has also been isolated from a cow,
while C. andersoni
is morphologically close to C. muris but infects cattle rather than mice. And
C. parvum includes a
complex of subspecies that specifically infect cattle, pigs,
kangaroos, ferrets or monkeys.
advance of molecular techniques, knowledge of the epidemiology
of human cryptosporidiosis has significantly increased. It has
been shown that the vast majority of human cases are caused by
(synonymous with C. parvum
genotype 1) and C. parvum
(synonymous with C. parvum
genotype 2). Other species, including
C. felis, C. canis
and C. muris can
also infect humans and are linked to clinical disease, not only
in immunocompromised patients but also in immunocompetent
Cryptosporidium serpentis is one of the most important
health concerns in snakes. The infection is characterized by
chronic clinical or subclinical symptoms. The presence of
hypertrophic gastritis, food regurgitation, progressive weight
loss, mortality, and intermittent or continuous shedding of
oocysts in the feces are some of the possible outcomes of
infection. There is no evidence that Cryptosporidium
serpentis is transmissible to humans or other mammals.
Although the life cycle of C. serpentis is not completely
understood, it is known that there are two infective stages of
the parasite. The first is a thick-walled oocyst which contains
four sporozoites that are released when the oocyst is ingested
by a snake. It is believed that the ingested oocysts invade a
small number of endothelial cells lining the stomach wall.
In these endothelial cells, the parasites undergo asexual
multiplication (schizogony or merogony) and then sexual
multiplication (gametogony), producing microgamonts and
macrogamonts. Upon fertilization of the macrogamonts by the
microgametes, oocysts develop and sporulate in the infected
host. Two different types of oocysts are produced: the
thick-walled, which is commonly excreted by the host after
sporogony, and the thin-walled oocyst, which is primarily
involved in autoinfection.
The thick-walled oocysts which are passed in the snake’s feces can remain
infective in the environment for months. These oocysts are
extremely resistant to temperature extremes and disinfectants.
As with most protozoan infections, this parasite can be acquired
by exposure to sporulated oocysts in contaminated food and water
and unhygienic enclosures. This microscopic parasite is tiny,
with oocysts typically measuring only 4-8 um in diameter.
Because of their small size, it is very hard to detect the
oocysts through microscopic examination.
Molecular detection by polymerase chain reaction can significantly
enhance the sensitivity and specificity of detection of
Cryptosporidium (da Silva et al., 2014). Since oocysts are
only intermittently shed, multiple samples can be taken at
intervals to increase the likelihood of detection. Useful sample
types for this purpose include feces, gastric lavage, endoscopic
gastric biopsy, fecal smears, and smears of mucous adhered to
regurgitated prey items
PCR is a rapid and
extremely sensitive technique for the detection of
Help confirm the disease causing agent
Shorten the time required to confirm a clinical
Help ensure that animal groups and facilities are free of
Early prevention of spread of this protozoan
Minimize human exposure to this protozoan
Thomas, A.L. and Chalmers, R.M. (2003) Investigation of the
range of Cryptosporidium
species detected by commercially available antibody-based tests.
Proceedings of the Health Protection Agency Inaugural
Conference, Warwick, September.
da Silva, D.C., Paiva, P.R., Nakamura, A.A., Homem, C.G., de Souza, M.S.,
Grego, K.F. and Meireles,
M.V. (2014) The detection of Cryptosporidium serpentis in snake
fecal samples by real-time PCR. Vet. Parasitol. 204:134-138.
Specimen requirements: 0.2 ml feces, or rectal swab,
or 0.2 ml gastric lavage, or swab of mucous adhered to
regurgitated prey items; or 0.2 ml fresh, frozen or fixed
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