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Zoologix performs environmental, zoo, wildlife and aquatic PCR tests for...

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Cryptosporidium PCR test
environmental, wildlife and zoo assay data sheet

Cryptosporidium

Test codes:

P0008 - Ultrasensitive qualitative screen for Cryptosporidium by real time polymerase chain reaction. This screen detects but does not differentiate Cryptosporidium species.

Test P0008 is included in P0041 - waterborne pathogens screening panel and in P0047 - ruminant fecal screening panel

X0033 - Ultrasensitive qualitative detection of Cryptosporidium serpentis by real time polymerase chain reaction.

 

Several of 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 Cryptosporidium species are currently accepted, on the basis of host specificity, pathogenesis, morphology and genotyping:

Infecting mammals: Cryptosporidium hominis, C. parvum, C. wrairi, C. felis, C. canis, C. andersoni, C. muris and C. ubiquitum

Infecting birds: C. baileyi, C. meleagridis and C. galli

Infecting reptiles: C. serpentis and C. saurophilum

Infecting fish: C. molnari

Phylogenetic 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 various Cryptosporidium 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.

With the 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 C. hominis (synonymous with C. parvum genotype 1) and C. parvum (synonymous with C. parvum genotype 2). Other species, including C. meleagridis, 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 people.

Cryptosporidium serpentis
Infection by 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 Cryptosporidium.

Utilities:

  • Help confirm the disease causing agent
  • Shorten the time required to confirm a clinical diagnosis of Cryptosporidium infection.
  • Help ensure that animal groups and facilities are free of Cryptosporidium
  • Early prevention of spread of this protozoan
  • Minimize human exposure to this protozoan

References:
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 gastric tissue.

For specimen types other than those listed here, please call to confirm specimen acceptability and shipping instructions.

For all 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

Turnaround time: 2 business days

Methodology: Qualitative real time PCR

Normal range: Nondetected

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