Downloaded from www.microbiologyresearch.org by IP: 54.162.190.106 On: Wed, 17 Feb 2016 06:54:18 Campylobacter canadensis sp. nov., from captive whooping cranes in Canada G. Douglas Inglis, 1 Bryanne M. Hoar, 2 Douglas P. Whiteside 2,3,4 and Douglas W. Morck 2,3 Correspondence G. Douglas Inglis inglisd@agr.gc.ca 1 Food Safety and Quality National Program, Agriculture and Agri-Food Canada Research Centre, 5403 1st Avenue S, Lethbridge, AB, T1J 4B1, Canada 2 Department of Biological Sciences, Faculty of Science, University of Calgary, 2500 University Drive NW, Calgary, AB, T2N 1N4, Canada 3 Faculty of Veterinary Medicine, University of Calgary, 2500 University Drive NW, Calgary, AB, T2N 1N4, Canada 4 Calgary Zoo Animal Health Centre, 1625 Centre Avenue E, Calgary, AB, T2E 8K2, Canada Ten isolates of an unknown Campylobacter species were isolated from cloacal swabs obtained from captive adult whooping cranes (Grus americana). All isolates were identified as Campylobacter based on generic PCR and grouped with other Campylobacter species based on 23S rRNA gene sequence. None of the isolates could be identified by species-specific PCR for known taxa, and all ten isolates formed a robust clade that was very distinct from known Campylobacter species based on 16S rRNA, rpoB and cpn60 gene sequences. The results of 16S rRNA gene nucleotide sequence (¡92 % sequence similarity to recognized Campylobacter species) and genomic DNA (no detectable relatedness) analyses were consistent with novel species status. Cells of the Campylobacter from whooping cranes were uniflagellar and typically sigmoid to allantoid in shape (0.48 mm wide and 2.61 mm long), but also spheroid to coccoid (0.59 mm wide and 0.73 mm long). The bacterium was oxidase-positive, able to reduce nitrite, able to grow at 376 and 42 6C, and grew anaerobically, as well as in an atmosphere devoid of H 2 , and on MacConkey agar. It was not a-haemolytic and was negative for hippurate and indoxyl acetate hydrolysis and alkaline phosphatase. It also was susceptible to cephalotin and was unable to grow on nutrient agar, on a medium containing 3.5 % NaCl or in ambient O 2 . The bacterium was unable to grow at 25 6C and growth was negative or very restricted at 30 6C. Fluorescent amplified fragment length polymorphism analysis indicated that nine of the recovered isolates were genetically distinct. A species-specific primer set targeting the cpn60 gene was developed. The name Campylobacter canadensis sp. nov. is proposed for the novel species, with the type strain L266 T (5CCUG 54429 T 5LMG 24001 T ). Ten isolates of a Campylobacter-like bacterium were recovered from captive whooping cranes (Grus americana), a bird whose populations have been so decimated by human activities that the species is currently classified as endangered in both Canada and the USA. In this paper, we describe the cultural and biochemical characteristics of this bacterium and the results of a polyphasic taxonomic investigation. Three unidentified isolates of a Campylobacter-like bac- terium were recovered from cloacae of three whooping cranes in 2004 during a study to characterize the normal enteric bacterial flora of captive whooping and sandhill cranes at the Calgary Zoo (Hoar et al., 2007). Seven additional isolates were recovered from whooping cranes in 2006. Isolations were made on a Campylobacter-selective Preston medium containing selective supplement SR117 (Oxoid), Karmali agar containing selective supplement SR167 (Oxoid) or Campylobacter charcoal-deoxycholate medium (CCDA) containing selective supplement SR155 (Oxoid) incubated anaerobically (10 % CO 2 , 10 % H 2 and 80 % N 2 ) or microaerobically (5 % O 2 , 10 % CO 2 ,3%H 2 Abbreviation: AFLP, amplified fragment length polymorphism. The GenBank/EMBL/DDBJ accession numbers for the 23S rRNA, 16S rRNA, rpoB and cpn60 gene sequences of the novel isolates determined in this study are EF621904 and EF621905 (23S rRNA), EF621894– EF621903 (16S rRNA), EF621885–EF621893 (rpoB) and EF621906–EF621915 (cpn60), as detailed in Figs 2–5; the accession numbers for strain L266 T are respectively EF621904, EF621894, EF621885 and EF621906. International Journal of Systematic and Evolutionary Microbiology (2007), 57, 2636–2644 DOI 10.1099/ijs.0.65061-0 2636 65061 G 2007 IUMS Printed in Great Britain