Hostname: page-component-cd9895bd7-gbm5v Total loading time: 0 Render date: 2024-12-23T07:43:40.208Z Has data issue: false hasContentIssue false

Molecular identification of Sarcocystis spp. helped to define the origin of green pythons (Morelia viridis) confiscated in Germany

Published online by Cambridge University Press:  05 December 2013

GASTÓN MORÉ*
Affiliation:
Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Epidemiology, Südufer 10, 17493 Greifswald-Insel Riems, Germany Laboratorio de Inmunoparasitología, Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, 60 y 118, 1900 La Plata, Argentina Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), C1033AAJ Buenos Aires, Argentina
NIKOLA PANTCHEV
Affiliation:
IDEXX Vet Med Lab, 71636 Ludwigsburg, Germany
DALAND C. HERRMANN
Affiliation:
Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Epidemiology, Südufer 10, 17493 Greifswald-Insel Riems, Germany
MAJDA GLOBOKAR VRHOVEC
Affiliation:
IDEXX Vet Med Lab, 71636 Ludwigsburg, Germany
SABINE ÖFNER
Affiliation:
Reptile Rescue Centre Munich, 80539 Munich, Germany
FRANZ J. CONRATHS
Affiliation:
Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Epidemiology, Südufer 10, 17493 Greifswald-Insel Riems, Germany
GEREON SCHARES
Affiliation:
Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Epidemiology, Südufer 10, 17493 Greifswald-Insel Riems, Germany
*
* Corresponding author: Laboratorio de Inmunoparasitología, Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, calle 60 y 118, 1900 La Plata, Argentina. E-mail: [email protected]

Summary

Sarcocystis spp. represent apicomplexan parasites. They usually have a heteroxenous life cycle. Around 200 species have been described, affecting a wide range of animals worldwide, including reptiles. In recent years, large numbers of reptiles have been imported into Europe as pets and, as a consequence, animal welfare and species protection issues emerged. A sample of pooled feces from four confiscated green pythons (Morelia viridis) containing Sarcocystis spp. sporocysts was investigated. These snakes were imported for the pet trade and declared as being captive-bred. Full length 18S rRNA genes were amplified, cloned into plasmids and sequenced. Two different Sarcocystis spp. sequences were identified and registered as Sarcocystis sp. from M. viridis in GenBank. Both showed a 95–97% sequence identity with the 18S rRNA gene of Sarcocystis singaporensis. Phylogenetic analysis positioned these sequences together with other Sarcocystis spp. from snakes and rodents as definitive and intermediate hosts (IH), respectively. Sequence data and also the results of clinical and parasitological examinations suggest that the snakes were definitive hosts for Sarcocystis spp. that circulate in wild IH. Thus, it seems unlikely that the infected snakes had been legally bred. Our research shows that information on the infection of snakes with Sarcocystis spp. may be used to assess compliance with regulations on the trade with wildlife species.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2013 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Barta, J. R. (1989). Phylogenetic analysis of the class Sporozoea (phylum Apicomplexa Levine, 1970): evidence for the independent evolution of heteroxenous life cycles. Journal of Parasitology 75, 195206.CrossRefGoogle ScholarPubMed
Carreno, R. A., Schnitzler, B. E., Jeffries, A. C., Tenter, A. M., Johnson, A. M. and Barta, J. R. (1998). Phylogenetic analysis of coccidia based on 18S rDNA sequence comparison indicates that Isospora is most closely related to Toxoplasma and Neospora . Journal of Eukaryotic Microbiology 45, 184188.CrossRefGoogle ScholarPubMed
Dahlgren, S. S. and Gjerde, B. (2007). Genetic characterisation of six Sarcocystis species from reindeer (Rangifer tarandus tarandus) in Norway based on the small subunit rRNA gene. Veterinary Parasitology 146, 204213.CrossRefGoogle ScholarPubMed
Dahlgren, S. S., Gouveia-Oliveira, R. and Gjerde, B. (2008). Phylogenetic relationships between Sarcocystis species from reindeer and other Sarcocystidae deduced from ssu rRNA gene sequences. Veterinary Parasitology 151, 2735.CrossRefGoogle ScholarPubMed
Dolezel, D., Koudela, B., Jirku, M., Hypsa, V., Obornik, M., Votypka, J., Modry, D., Slapeta, J. R. and Lukes, J. (1999). Phylogenetic analysis of Sarcocystis spp. of mammals and reptiles supports the coevolution of Sarcocystis spp. with their final hosts. International Journal for Parasitology 29, 795798.CrossRefGoogle ScholarPubMed
Drummond, A. J., Ashton, B., Buxton, S., Cheung, M., Cooper, A., Heled, J., Kearse, M., Moir, R., Stones-Havas, S., Sturrock, S., Thierer, T. and Wilson, A. (2011). Geneious v5.5. http://www.geneious.com/.Google Scholar
Dubey, J. P., Speer, C. A. and Fayer, R. (1989). Sarcocystosis of Animals and Man, 1st Edn. CRC Press, Boca Raton, FL, USA.Google Scholar
Duszynski, D. W. and Upton, S. J. (2009). The Biology of the Coccidia (Apicomplexa) of Snakes of the World, 1st Edn. CreateSpace, USA.Google Scholar
Fenger, C. K., Granstrom, D. E., Langemeier, J. L., Stamper, S., Donahue, J. M., Patterson, J. S., Gajadhar, A. A., Marteniuk, J. V., Xiaomin, Z. and Dubey, J. P. (1995). Identification of opossums (Didelphis virginiana) as the putative definitive host of Sarcocystis neurona . Journal of Parasitology 81, 916919.CrossRefGoogle ScholarPubMed
Herrmann, D. C., Maksimov, A., Pantchev, N., Vrhovec, M. G., Conraths, F. J. and Schares, G. (2011). Comparison of different commercial DNA extraction kits to detect Toxoplasma gondii oocysts in cat faeces. Berliner und Münchener tierärztliche Wochenschrift 124, 497502.Google ScholarPubMed
Ho, M. S., Barr, B. C., Marsh, A. E., Anderson, M. L., Rowe, J. D., Tarantal, A. F., Hendrickx, A. G., Sverlow, K., Dubey, J. P. and Conrad, P. A. (1996). Identification of bovine Neospora parasites by PCR amplification and specific small-subunit rRNA sequence probe hybridization. Journal of Clinical Microbiology 34, 12031208.CrossRefGoogle ScholarPubMed
Jeffries, A. C., Schnitzler, B., Heydorn, A. O., Johnson, A. M. and Tenter, A. M. (1997). Identification of synapomorphic characters in the genus Sarcocystis based on 18S rDNA sequence comparison. Journal of Eukaryotic Microbiology 44, 388392.CrossRefGoogle ScholarPubMed
Jinnai, M., Kawabuchi-Kurata, T., Tsuji, M., Nakajima, R., Fujisawa, K., Nagata, S., Koide, H., Matoba, Y., Asakawa, M., Takahashi, K. and Ishihara, C. (2009). Molecular evidence for the presence of new Babesia species in feral raccoons (Procyon lotor) in Hokkaido, Japan. Veterinary Parasitology 162, 241247.CrossRefGoogle ScholarPubMed
Lyons, J. A. and Natusch, D. J. D. (2011). Wildlife laundering through breeding farms: illegal harvest, population declines and a means of regulating the trade of green pythons (Morelia viridis) from Indonesia. Biological Conservation 144, 30733081.CrossRefGoogle Scholar
Matuschka, F. R. (1987). Reptiles as intermediate and/or final hosts of Sarcosporidia. Parasitology Research 73, 2232.CrossRefGoogle ScholarPubMed
McAllister, C. T., Upton, S. J., Trauth, S. E. and Dixon, J. R. (1995). Coccidian parasites (Apicomplexa) from snakes in the southcentral and southwestern United States: new host and geographic records. Journal of Parasitology 81, 6368.CrossRefGoogle ScholarPubMed
Moré, G., Abrahamovich, P., Jurado, S., Bacigalupe, D., Marin, J. C., Rambeaud, M., Venturini, L. and Venturini, M. C. (2011). Prevalence of Sarcocystis spp. in Argentinean cattle. Veterinary Parasitology 177, 162165.CrossRefGoogle ScholarPubMed
Moré, G., Schares, S., Maksimov, A., Conraths, F. J., Venturini, M. C. and Schares, G. (2013). New real time PCR to differentiate Sarcocystis spp. affecting cattle. Veterinary Parasitology 197, 8594. doi: 10.1016/j.vetpar.2013.04.024.CrossRefGoogle Scholar
Mugridge, N. B., Morrison, D. A., Jakel, T., Heckeroth, A. R., Tenter, A. M. and Johnson, A. M. (2000). Effects of sequence alignment and structural domains of ribosomal DNA on phylogeny reconstruction for the protozoan family sarcocystidae. Molecular Biology and Evolution 17, 18421853.CrossRefGoogle ScholarPubMed
Nijman, V. and Sheperd, C. R. (2009). Wildlife Trade from ASEAN to the EU: Issues with the Trade in Captive-Bred Reptiles from Indonesia. TRAFFIC Europe Report for the European Commission, Brussels, Belgium.Google Scholar
Odening, K. (1998). The present state of species-systematics in Sarcocystis Lankester, 1882 (Protista, Sporozoa, Coccidia). Systematic Parasitology 41, 209233.CrossRefGoogle Scholar
Pasmans, F., Blahak, S., Martel, A. and Pantchev, N. (2008). Introducing reptiles into a captive collection: the role of the veterinarian. Veterinary Journal 175, 5368.CrossRefGoogle ScholarPubMed
Schares, G., Heydorn, A. O., Cuppers, A., Mehlhorn, H., Geue, L., Peters, M. and Conraths, F. J. (2002). In contrast to dogs, red foxes (Vulpes vulpes) did not shed Neospora caninum upon feeding of intermediate host tissues. Parasitology Research 88, 4452.CrossRefGoogle Scholar
Schares, G., Pantchev, N., Barutzki, D., Heydorn, A. O., Bauer, C. and Conraths, F. J. (2005). Oocysts of Neospora caninum, Hammondia heydorni, Toxoplasma gondii and Hammondia hammondi in faeces collected from dogs in Germany. International Journal for Parasitology 35, 15251537.CrossRefGoogle ScholarPubMed
Slapeta, J. R., Modry, D. and Koudela, B. (1999). Sarcocystis atheridis sp. nov., a new sarcosporidian coccidium from Nitsche's bush viper, Atheris nitschei Tornier, 1902, from Uganda. Parasitology Research 85, 758764.Google Scholar
Slapeta, J. R., Modry, D., Votypka, J., Jirku, M., Koudela, B. and Lukes, J. (2001). Multiple origin of the dihomoxenous life cycle in sarcosporidia. International Journal for Parasitology 31, 413417.CrossRefGoogle ScholarPubMed
Slapeta, J. R., Kyselova, I., Richardson, A. O., Modry, D. and Lukes, J. (2002). Phylogeny and sequence variability of the Sarcocystis singaporensis Zaman and Colley, (1975) 1976 ssrDNA. Parasitology Research 88, 810815.Google ScholarPubMed
Slapeta, J. R., Modry, D., Votypka, J., Jirku, M., Lukes, J. and Koudela, B. (2003). Evolutionary relationships among cyst-forming coccidia Sarcocystis spp. (Alveolata: Apicomplexa: Coccidea) in endemic African tree vipers and perspective for evolution of heteroxenous life cycle. Molecular Phylogenetics and Evolution 27, 464475.CrossRefGoogle ScholarPubMed
Tenter, A. M., Baverstock, P. R. and Johnson, A. M. (1992). Phylogenetic relationships of Sarcocystis species from sheep, goats, cattle and mice based on ribosomal RNA sequences. International Journal for Parasitology 22, 503513.CrossRefGoogle ScholarPubMed
Tian, M., Chen, Y., Wu, L., Rosenthal, B. M., Liu, X., He, Y., Dunams, D. B., Cui, L. and Yang, Z. (2012). Phylogenetic analysis of Sarcocystis nesbitti (Coccidia: Sarcocystidae) suggests a snake as its probable definitive host. Veterinary Parasitology 183, 373376.CrossRefGoogle ScholarPubMed
Yang, Z. Q., Zuo, Y. X., Yao, Y. G., Chen, X. W., Yang, G. C. and Zhang, Y. P. (2001). Analysis of the 18S rRNA genes of Sarcocystis species suggests that the morphologically similar organisms from cattle and water buffalo should be considered the same species. Molecular and Biochemical Parasitology 115, 283288.CrossRefGoogle ScholarPubMed