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Appl Environ Microbiol, July 1998, p. 2736-2738, Vol. 64, No. 7
Department of Molecular Microbiology
and Immunology, School of Hygiene and Public Health, Johns Hopkins
University, Baltimore, Maryland 212051;
Immunity and Disease Resistance Laboratory, Livestock and
Poultry Science Institute, Agricultural Research Services, U.S.
Department of Agriculture, Beltsville, Maryland
207052;
Cooperative Oxford Laboratory,
National Marine Fisheries Service, National Oceanic and Atmospheric
Administration, U.S. Department of Commerce, Oxford, Maryland
216543; and
Department of Parasitic
Diseases, National Center for Infectious Diseases, Centers for
Disease Control and Prevention, Chamblee, Georgia
303414
Received 18 March 1998/Accepted 21 April 1998
Fecal droppings of migratory Canada geese, Branta
canadensis, collected from nine sites near the Chesapeake Bay
(Maryland), were examined for the presence of Cryptosporidium
parvum and Giardia spp. Cryptosporidium
sp. oocysts were found in feces at seven of nine sites, and
Giardia cysts were found at all nine sites. The oocysts
from three sites were infectious for mice and molecularly identified as
the zoonotic genotype of Cryptosporidium parvum. Waterfowl
can disseminate infectious C. parvum oocysts in the environment.
Two genotypes of
Cryptosporidium parvum, the animal-adapted (or zoonotic) and
human-adapted genotypes, have been identified by molecular
techniques (1, 12, 14); both genotypes can produce
life-threatening infections in immunocompromised and immunosuppressed people. The infectivity of Cryptosporidium oocysts is
long lasting (3). Aquatic birds have been incriminated
in the contamination of water with microbiological agents
(11, 16). Oral inoculations of Canada geese (Branta
canadensis) and ducks (Anas platyrhynchos) with
infectious C. parvum oocysts demonstrated that the oocysts passed through the gastrointestinal tract, retaining their infectivity (8, 10). This suggested that waterfowl had the potential to
serve as mechanical carriers of C. parvum and to disseminate infectious oocysts in the environment (2).
Giardia spp. are protozoan pathogens of
vertebrates; their infectious stage, the cyst, is
transmitted via water and fecal-oral contamination
(18). Successful transmission of avian isolates of
Giardia to mammals may have serious implications for
contamination of watersheds (17). The purpose of the present
study was to determine if Cryptosporidium oocysts or
Giardia cysts might be present in the fecal droppings of
free-ranging Canada geese, and if so, to assess the concentration of
these cystic stages in the goose feces. An additional goal was to
determine the infectivity for mammals of any
Cryptosporidium oocysts that were recovered from feces of
geese, including molecular characterization of the genotype.
Freshly deposited fecal droppings of B. canadensis were
collected from nine locations of the Chesapeake Bay (Maryland)
area (Table 1) between October and
November 1997. Each of nine pooled droppings from a single flock of
birds was weighed. Each of the collective fecal specimens was
separately processed (5). The fecal sediments (5)
were subjected to cesium chloride gradient centrifugation
(5) and processed by the cellulose acetate membrane-filter dissolution method for recovery of Cryptosporidium oocysts
(7). The method has a recovery efficiency of 78.8%
(9) and does not alter the infectivity of C. parvum oocysts (9). Giardia sp. cysts and
Cryptosporidium sp. oocysts were visualized with the
immunofluorescent antibody of the MERIFLUOR
Cryptosporidium/Giardia test kit (Meridian Diagnostics,
Cincinnati, Ohio) and counted. Equal fractions of 3 suspensions of
Cryptosporidium sp. oocysts recovered from feces collected
at Wye Island (Table 1) were pooled, and each of four 9-day-old
suckling BALB/c mice was inoculated with approximately 9.0 × 104 oocysts (3). The mice were euthanatized
96 h postinoculation, and their intestines were processed
(3). Histologic sections were examined for developmental
stages of Cryptosporidium (3). The remainder of
the oocyst pool was subjected to molecular genotyping utilizing PCR for
Cryptosporidium TRAP C2 and beta-tubulin genes (14).
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Giardia sp. Cysts and Infectious Cryptosporidium
parvum Oocysts in the Feces of Migratory Canada Geese
(Branta canadensis)
ABSTRACT
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TABLE 1.
Numbers and concentrations of Giardia sp.
cysts, Cryptosporidium sp. oocysts, and infectious C. parvuma oocysts recovered from fecal droppings
collected in the Chesapeake Bay (Maryland) area from nine flocks of
migratory Canada geese (B. canadiensis)
The mean weight of an individual fecal dropping ± standard deviation was 17.2 ± 1.9 g (range, 13.5 to 21.6 g). The total weight of feces collected was 12.6 kg (approximately 733 droppings). Oocysts of Cryptosporidium sp. were recovered from goose feces at seven of the nine sites (Table 1). Oocysts of Cryptosporidium sp. recovered from feces collected at Wye Island (Table 1) induced severe infections in all four mice. Results of PCR confirmed that the oocysts used in the mouse bioassay were of genotype 2, the animal-adapted, or zoonotic, genotype of C. parvum. The mean concentration of oocysts ± standard deviation was 370 ± 197 oocysts/g (Table 1). Giardia sp. cysts were recovered from all nine pooled fecal specimens. The concentration of Giardia sp. cysts was 405 cysts/g (Table 1). Comparison of the paired numbers of Giardia sp. cysts with Cryptosporidium oocysts in feces showed that the concentration of Giardia sp. cysts was significantly higher than the concentration of Cryptosporidium oocysts (Sign test; P < 0.02). The concentrations of cystic stages of both pathogens were correlated with each other (Spearman rank correlation; r = 0.69; P < 0.01). Significant correlation was observed between the weight of fecal specimens and the number of recovered oocysts (Spearman rank correlation; r = 0.50; P < 0.04).
Visitation of gulls at drinking and recreational water reservoirs caused a decline in the microbiological quality of such waters (11). Although viable oocysts of Cryptosporidium sp. were recovered from gull feces the organism was not identified to the species level (16). Gulls can be infected with Cryptosporidium baileyi (13). The species of oocysts recovered previously from gull droppings (16) is unknown, and therefore the epidemiological importance of gulls in transmitting C. parvum remains unknown. In contrast, the present study demonstrated for the first time the presence of infectious oocysts of C. parvum in the fecal droppings of birds, migratory Canada geese, as well as the presence of Giardia sp. cysts in these feces. Cryptosporidium parvum is unable to establish intestinal infection in birds (8, 10), and therefore we conclude that the infectious C. parvum oocysts were acquired by the birds from their natural habitat. The mouse bioassay and PCR showed that the oocysts recovered from goose feces collected at Wye Island (Table 1) were viable and infectious and represented the zoonotic genotype of C. parvum. Interestingly, the same genotype of C. parvum has been recovered from oysters in the Wye River near Wye Island (3). The Eastern Shore of Maryland is a predominantly agricultural region with scattered cattle farms, and migratory geese were actually observed to wander behind the cattle and pick up undigested corn from their feces. Previous studies showed that approximately 25% of C. parvum oocysts administered orally to geese and ducks was recovered (8, 10); thus, geese in the present study may actually ingest higher numbers of the oocysts than we recovered. Because avian giardiasis produces a minimal number of cysts (4) and this pathogen was not previously reported from the Canada goose, it is possible that Giardia cysts were acquired by the birds in their natural habitat. It has been suggested that aquatic birds such as herons be considered as reservoirs for human waterborne giardiasis (4). Avian isolates of Giardia cysts established severe infections in mice, and it has been concluded that birds can contaminate waterways with Giardia cysts and that these cysts are potentially a danger for humans, livestock, and native animals (17). Irrespective of the origin of Giardia sp. cysts in the goose feces (indigenous infection versus a mechanical carrier), these cysts produced positive immunofluorescent-antibody reactions with the commercial test kit used for testing of raw water samples (6, 15). The present study provides clear evidence that birds can act as mechanical carriers of infectious oocysts of C. parvum and can disseminate these oocysts in the environment, including drinking water supplies.
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ACKNOWLEDGMENTS |
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We thank the Wye Island Natural Resources Management Area (Queenstown, Md.) and the Wildfowl Trust of North America (Grasonville, Md.) for facilitating collection of feces of migratory Canada geese and K. N. Greenhawk (COLMAP, Oxford, Md.) for providing geographical location data. We acknowledge C. Carpenter for technical assistance.
This study was supported in part by Maryland Sea Grant R/F-88.
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FOOTNOTES |
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* Corresponding author. Mailing address: Department of Molecular Microbiology and Immunology, School of Hygiene and Public Health, Johns Hopkins University, Baltimore, MD 21205. Phone: (410) 614-4984. Fax: (410) 955-0105. E-mail: tgraczyk{at}jhsph.edu.
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REFERENCES |
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Text
References
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| 1. | Awad-El-Kariem, F. M., H. A. Robinson, D. A. Dyson, D. Evans, S. Wright, M. T. Fox, and V. M. McDonald. 1995. Differentiation between human and animal strains of Cryptosporidium parvum using isoenzyme typing. Am. J. Trop. Med. Hyg. 52:559-564. |
| 2. | Fayer, R., T. K. Graczyk, C. A. Farley, E. J. Lewis, and J. M. Trout. 1997. The potential role of waterfowl and oysters in the complex epidemiology of Cryptosporidium parvum, p. 153-158. In C. R. Fricker, J. L. Clancy, and P. A. Rochelle (ed.), 1997 International Symposium on Waterborne Cryptosporidium proceedings. American Water Works Association Press, Denver, Colo. |
| 3. |
Fayer, R.,
T. K. Graczyk,
E. J. Lewis,
J. M. Trout, and C. A. Farley.
1998.
Survival of infectious Cryptosporidium parvum oocysts in seawater and Eastern oysters (Crassostrea virginica) in the Chesapeake Bay.
Appl. Environ. Microbiol.
64:1070-1074 |
| 4. |
Georgi, M. E.,
M. S. Carlisle, and L. E. Smiley.
1986.
Giardiasis in a great blue heron (Ardea herodias) in New York state: another potential source of waterborne giardiasis.
Am. J. Epidemiol.
123:916-917 |
| 5. | Graczyk, T. K., and M. R. Cranfield. 1997. Assessment of the conventional detection of fecal Cryptosporidium serpentis oocysts in subclinically infected captive snakes. Vet. Res. 27:185-192. |
| 6. | Graczyk, T. K., M. R. Cranfield, and R. Fayer. 1996. Evaluation of commercial immunoassay (EIA) and immunofluorescent antibody (IFA) test kit for detection of Cryptosporidium oocysts other than Cryptosporidium parvum. Am. J. Trop. Med. Hyg. 54:274-279. |
| 7. | Graczyk, T. K., M. R. Cranfield, and R. Fayer. 1997. Recovery of waterborne oocysts of Cryptosporidium from water samples by the membrane-filter dissolution method. Parasitol. Res. 83:121-125[Medline]. |
| 8. | Graczyk, T. K., M. R. Cranfield, R. Fayer, and M. S. Anderson. 1996. Viability and infectivity of Cryptosporidium parvum oocysts are retained upon intestinal passage through a refractory avian host. Appl. Environ. Microbiol. 62:3234-3237[Abstract]. |
| 9. | Graczyk, T. K., R. Fayer, M. R. Cranfield, and R. Owens. 1997. Cryptosporidium parvum oocysts recovered from water by the membrane filter dissolution method retain their infectivity. J. Parasitol. 83:111-114[Medline]. |
| 10. | Graczyk, T. K., M. R. Cranfield, R. Fayer, J. Trout, and H. J. Goodale. 1997. Infectivity of Cryptosporidium parvum oocysts is retained upon intestinal passage through a migratory waterfowl species (Canada goose, Branta canadensis). Trop. Med. Int. Health 2:341-347. [Medline] |
| 11. | Hatch, J. J. 1996. Threats to public health from gulls (Laridae). Int. J. Environ. Health Res. 6:5-16. |
| 12. | Morgan, U. M., C. C. Constantine, P. O'Donoghue, B. P. Meloni, P. A. O'Brien, and R. C. A. Thompson. 1995. Molecular characterization of Cryptosporidium isolates from humans and other animals using random amplified polymorphic DNA analysis. Am. J. Trop. Med. Hyg. 52:559-564. |
| 13. | Pavlasek, I. 1993. The black-headed gull (Larus ridibundus L.), a new host for Cryptosporidium baileyi (Apicomplexa: Cryptosporidiidae). Vet. Med. 38:629-638. |
| 14. | Peng, M. M., L. Xiao, A. R. Freeman, M. J. Arrowood, A. A. Escalante, A. C. Weltman, C. S. L. Ong, W. R. MacKenzie, A. A. Lal, and C. B. Beard. 1997. Genetic polymorphism among Cryptosporidium parvum isolates: evidence for two distinct human transmission cycles. Emerg. Infect. Dis. 3:567-573. [Medline] |
| 15. | Smith, H. V., and J. B. Rose. 1998. Waterborne cryptosporidiosis: current status. Parasitol. Today 14:14-22. |
| 16. | Smith, H. V., J. Brown, J. C. Coulson, G. P. Morris, and R. W. A. Girdwood. 1993. Occurrence of oocysts of Cryptosporidium sp. in Larus spp. gulls. Epidemiol. Infect. 110:135-143[Medline]. |
| 17. | Upcroft, J. A., P. A. McDonnell, A. N. Gallagher, N. Cheng, and P. Upcroft. 1997. Lethal Giardia from a wild-caught sulphur-crested cockatoo (Cacatus galerita) established in vitro chronically infects mice. Parasitology 114:407-412. |
| 18. |
Wolfe, M. S.
1992.
Giardiasis.
Clin. Microbiol. Rev.
5:93-100 |
Appl Environ Microbiol, July 1998, p. 2736-2738, Vol. 64, No. 7
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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