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Volume 15, Number 5—May 2009
Letter

Campylobacter jejuni in Penguins, Antarctica

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To the Editor: The wildlife of Antarctica is highly specialized. Although large animal species are limited primarily to penguins and seals, each species is often abundant. The high degree of isolation potentially protects Antarctic wildlife from diseases distributed in other areas of the world (1,2). Despite Antarctica’s isolation, however, human- or animal-related pathogens have been found there, or in the sub-Antarctic islands. For instance, serologic evidence of influenza virus A infections in penguins has been found (3), and both Salmonella spp. and Mycobacterium tuberculosis have been isolated from sub-Antarctic and Antarctic animals (4,5).

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Thumbnail of Macaroni penguins (Eudyptes chrysolophus). Photo by Jonas Bonnedahl.

Figure. Macaroni penguins (Eudyptes chrysolophus). Photo by Jonas Bonnedahl.

Campylobacter jejuni is a leading cause of bacterial gastroenteritis in humans worldwide; it is usually found in the intestinal tract of various farm and wild animals, particularly birds (6,7).We previously reported finding 3 C. jejuni subsp. jejuni isolates in macaroni penguins (Eudyptes chrysolophus; Figure) from Bird Island (54°00′S, 38°02′W), South Georgia (1). Phenotypic tests and 16S rRNA gene sequencing showed that the penguin isolates were identical to each other, and macrorestriction profiling of pulsed-field gel electrophoresis fragments showed that they were very similar to fragments isolated from poultry in Washington in 1984 (1). Because the isolates were retrieved from macaroni penguin chicks, we concluded that the animals had acquired the infection locally and that this was likely an instance of introduction of a pathogen to the Antarctic region.

However, restriction fragment pattern resemblance is not identical to genetic relatedness and, given the relevance of the question of origin, this resemblance led us to use a new method for genetic characterization. We reanalyzed the macaroni penguin isolates with multilocus sequence typing (MLST), a method that uses sequence data from 7 unlinked loci for genetic identification (8), complemented with flaA gene sequencing. A benefit of this method is the increasing availability of epidemiologic databases in which isolates can be compared (e.g., http://pubmlst.org/cam,pylobacter). The isolates were thawed and cultured on conventional blood agar (Columbia agar II containing 8% [vol/vol] whole horse blood) at 42°C in a microaerobic gas environment, with the CampyGen gas-generating system (CN0025A; Oxoid Ltd, Basingstoke, UK) and the BBL GasPak system (BD, Franklin Lakes, NJ, USA). Bacterial DNA was prepared by making a suspension of freshly grown bacterial cells in 200 µL of phosphate-buffered saline (Sigma, St. Louis, MO, USA). Genomic DNA was extracted by use of a Bio Robot M48 (QIAGEN, Hilden, Germany) with a MagAttract DNA mini M48 kit, according to the instructions of the manufacturer. The PCR amplification and nucleotide sequencing followed the original protocol in principle (8). The amplification products were purified and sequenced by using internal separated nested primer pairs.

The 3 isolates from macaroni penguins were all of the same genotype (sequence type [ST]–45) and thus have a common origin. The ST-45 sequence type is the central genotype of the ST-45 clonal complex, a complex often associated with human disease and asymptomatic infection in poultry (9,10). Indeed, nearly 42% of the ST-45 samples available in the MLST database have been isolated from humans (31% from poultry), and similar percentages have been observed for the ST-45 clonal complex as a whole. The ST-45 clonal complex is large (composed of 195 individual STs) and has been isolated to date from a variety of environmental sources and different geographic regions, with the exception of the Arctic. The isolates were identical at the flaA locus, all having the allele 21/peptide 2 designation (http://pubmlst.org/campylobacter/flaA). This particular peptide is found in 31 records in the database and is thus not unique to the penguin isolates.

Our MLST analysis confirms that the C. jejuni isolates from the penguins were of a genotype common among humans with disease and among our food animals. C. jejuni is not normally distributed among Antarctic animals (1,2), which indicates that this strain may have been imported through human activities. On Bird Island, such activities were carried out by scientists at the British Antarctic Survey base. At the time of the study, toilet wastes from the station were emptied into the surrounding waters, providing a possible transmission route for human-associated C. jejuni to reach wildlife, including penguins. Other possible sources of the C. jejuni infections include wastes from passing ships or seabirds that pick up the bacteria during offshore feeding excursions (for albatrosses, these can be ≈1,000 km). Once established in a penguin colony, a gastrointestinal pathogen may be transmitted rapidly among individual birds as they are breeding densely and producing a large amount of feces (guano) in the colony. C. jejuni infection in birds is normally not associated with overt disease, but other and possibly more devastating pathogens introduced to Antarctic animals could potentially cause outbreaks.

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Acknowledgments

We thank T. Broman for laboratory assistance and comments.

This study was logistically supported by the Swedish Polar Research Institute and the British Antarctic Survey and gained financial support from the Swedish Research Council for Environment, Agricultural Sciences and Spatial Planning (FORMAS).

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Petra Griekspoor, Björn Olsen, and Jonas WaldenströmComments to Author 

Author affiliations: Kalmar University, Kalmar, Sweden (P. Griekspoor, B. Olsen, J. Waldenström); Uppsala University, Uppsala, Sweden (B. Olsen)

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References

  1. Broman  T, Bergstrom  S, On  SL, Palmgren  H, McCafferty  DJ, Sellin  M, Isolation and characterization of Campylobacter jejuni subsp. jejuni from macaroni penguins (Eudyptes chrysolophus) in the subantarctic region. Appl Environ Microbiol. 2000;66:44952. DOIPubMed
  2. Bonnedahl  J, Olsen  B, Waldenström  J, Broman  T, Jalava  J, Huovinen  P, Antibiotic susceptibility of faecal bacteria in Antarctic penguins. Polar Biol. 2008;31:75963. DOI
  3. Wallensten  A, Munster  V, Osterhaus  AD, Waldenström  J, Bonnedahl  J, Broman  T, Mounting evidence for the presence of influenza A virus in the avifauna of the Antarctic region. Antarct Sci. 2006;18:3536. DOI
  4. Palmgren  H, McCafferty  D, Aspan  A, Broman  T, Sellin  M, Wollin  R, Salmonella in sub-Antarctica: low heterogeneity in Salmonella serotypes in South Georgian seals and birds. Epidemiol Infect. 2000;125:25762. DOIPubMed
  5. Bastida  R, Loureiro  J, Quse  V, Bernardelli  A, Rodriguez  D, Costa  E. Tuberculosis in a wild subantarctic fur seal from Argentina. J Wildl Dis. 1999;35:7968.PubMed
  6. Miller  WG, Mandrell  RE. Prevalence of Campylobacter in the food and water supply: incidence, outbreaks, isolation and detection. In: Ketley JM, Konkel ME, editors. Campylobacter: molecular and cellular biology. Wymondham (UK): Horizon Bioscience; 2005. p. 101–63.
  7. Waldenström  J, Broman  T, Carlsson  I, Hasselquist  D, Achterberg  RP, Wagenaar  JA, Prevalence of Campylobacter jejuni, Campylobacter lari, and Campylobacter coli in different ecological guilds and taxa of migrating birds. Appl Environ Microbiol. 2002;68:59117. DOIPubMed
  8. Dingle  KE, Colles  FM, Wareing  DR, Ure  R, Fox  AJ, Bolton  FE, Multilocus sequence typing system for Campylobacter jejuni. J Clin Microbiol. 2001;39:1423. DOIPubMed
  9. Dingle  KE, Colles  FM, Ure  R, Wagenaar  JA, Duim  B, Bolton  FJ, Molecular characterization of Campylobacter jejuni clones: a basis for epidemiologic investigation. Emerg Infect Dis. 2002;8:94955.PubMed
  10. Manning  G, Dowson  CG, Bagnall  MC, Ahmed  IH, West  M, Newell  DG. Multilocus sequence typing for comparison of veterinary and human isolates of Campylobacter jejuni. Appl Environ Microbiol. 2003;69:63709. DOIPubMed

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DOI: 10.3201/eid1505.081160

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Table of Contents – Volume 15, Number 5—May 2009

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Jonas Waldenström, Section for Zoonotic Ecology and Epidemiology, School of Pure and Applied Natural Sciences, Kalmar University, SE 391, 82 Kalmar, Sweden

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