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Campylobacter ornithocola sp. nov., a new member of the Campylobacter lari group isolated from wild bird
faecal samples.
Alberto Caceres, Ivo Muñoz, Gregorio Iraola, Florencia Díaz-Viraqué, Luis Collado
To cite this version:
Alberto Caceres, Ivo Muñoz, Gregorio Iraola, Florencia Díaz-Viraqué, Luis Collado. Campylobacter
ornithocola sp. nov., a new member of the Campylobacter lari group isolated from wild bird faecal
samples.. International Journal of Systematic and Evolutionary Microbiology, Microbiology Society,
2017, 67 (6), pp.1643-1649. �10.1099/ijsem.0.001822�. �pasteur-01499037�
International Journal of Systematic and Evolutionary Microbiology
Campylobacter ornithocola sp. nov., a new member of the Campylobacter lari group isolated from wild bird faecal samples
--Manuscript Draft--
Manuscript Number: IJSEM-D-16-00613R2
Full Title: Campylobacter ornithocola sp. nov., a new member of the Campylobacter lari group isolated from wild bird faecal samples
Article Type: Note
Section/Category: New taxa - Proteobacteria
Keywords: Wild bird, Campylobacter, Valdivia, Chile
Corresponding Author: Luis Collado, Ph.D
Universidad Austral de Chile Valdivia, CHILE
First Author: Alberto Caceres
Order of Authors: Alberto Caceres
Ivo Muñoz Gregorio Iraola Florencia Díaz-Viraqué Luis Collado, Ph.D Manuscript Region of Origin: CHILE
Abstract: During a study on the prevalence and diversity of campylobacteria in wild birds faecal
samples in the city of Valdivia (South of Chile) 17 Gram-negative, curved rod isolates,
were initially identified as Campylobacter lari by PCR-RFLP. Further identification by
16S rRNA sequence analysis revealed that they formed a distinct group in the genus
Campylobacter. This unique position was confirmed by rpoB, atpA and cpn60 gene
sequence analysis. The average nucleotide identity between the representative strain
WBE38T and the type strain of the closest taxon Campylobacter lari subsp. concheus
(LMG 11760) was 90.8%. The oxidase and urease activity of the novel isolates
enabled them to be phenotypically differentiated from recognized Campylobacter
species. Therefore, on the basis of phenotypic, genetic and genomic characterizations,
this study clearly shows that these strains represent a novel species within the genus
Campylobacter, for which the name Campylobacter ornithocola sp. nov. is proposed,
with the type strain WBE38T (=CECT 9147T =LMG 29815T).
Campylobacter ornithocola sp. nov., a new member of the Campylobacter lari 1
group isolated from wild bird faecal samples 2
3
Alberto Cáceres
1, Ivo Muñoz
1, Gregorio Iraola
2, Florencia Díaz-Viraqué
3and Luis Collado
14
5
1
Institute of Biochemistry and Microbiology, Faculty of Sciences, Universidad Austral de 6
Chile, Valdivia, Chile.
2Bioinformatics Unit, Institut Pasteur Montevideo, Montevideo, 7
Uruguay.
3Molecular Biology Unit, Institut Pasteur Montevideo, Montevideo, Uruguay.
8
9
Running title: Campylobacter ornithocola sp. nov.
10
11
Corresponding Author:
12
Luis Collado González 13
Institute of Biochemistry and Microbiology 14
Faculty of Sciences 15
Universidad Austral de Chile 16
Isla Teja Campus, Valdivia, Chile 17
Phone: 56-63-2293848 18
Fax: 56-63-2215295 19
E-mail: luiscollado@uach.cl 20
Manuscript Including References (Word document) Click here to download Manuscript Including References (Word
document) C_ornithocola_sp_nov R2..doc
The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA, rpoB, atpA, cpn60 21
genes and the draft genome of the strain WBE38
Tare KX467974, KX467989, KX467979, 22
KX467985 and LXSU00000000, respectively.
23
24
25
ABSTRACT 26
During a study on the prevalence and diversity of campylobacteria in wild birds faecal 27
samples in the city of Valdivia (South of Chile) 17 Gram-negative, curved rod isolates, 28
were initially identified as Campylobacter lari by PCR-RFLP. Further identification by 16S 29
rRNA sequence analysis revealed that they formed a distinct group in the genus 30
Campylobacter. This unique position was confirmed by rpoB, atpA and cpn60 gene 31
sequence analysis. The average nucleotide identity between the representative strain 32
WBE38
Tand the type strain of the closest taxon Campylobacter lari subsp. concheus 33
(LMG 11760) was 90.8%. The oxidase and urease activity of the novel isolates enabled 34
them to be phenotypically differentiated from recognized Campylobacter species.
35
Therefore, on the basis of phenotypic, genetic and genomic characterizations, this study 36
clearly shows that these strains represent a novel species within the genus 37
Campylobacter, for which the name Campylobacter ornithocola sp. nov. is proposed, with 38
the type strain WBE38
T(=CECT 9147
T=LMG 29815
T).
39
40
41
42
43
The genus Campylobacter was proposed by Sebald & Veron (1963), to accommodate the 44
species Vibrio fetus (now Campylobacter fetus, the type species of the genus). Since then 45
the number of Campylobacter species has greatly increased and, at the time of writing 46
(September 2016), this genus encompassed 28 species and 9 subspecies with valid 47
published names (LPSN, 2016; Piccirillo et al., 2016; Van et al., 2016). The Campylobacter 48
infection is the leading cause of foodborne bacterial gastroenteritis in the developed world, 49
where infection is principally the result of consumption of contaminated meat and poultry.
50
The most common symptoms of campylobacteriosis include severe diarrhoea, with a 51
proportion of patients developing chronic sequelae such as Guillain-Barré syndrome, 52
reactive arthritis and irritable bowel syndrome (Man, 2011). Campylobacter jejuni and 53
Campylobacter coli are by far the most isolated and studied species, while the 54
epidemiology and the clinical role of the other less common species in human and animal 55
diseases is far less understood (Man, 2011; Iraola et al., 2014).
56
In the present study, 17 new Campylobacter isolates recovered from wild bird faecal 57
samples were subjected to a polyphasic approach including molecular identification by 58
PCR-RFLP, genotyping by ERIC-PCR, phylogenetic analysis using the 16S rRNA, rpoB, 59
atpA and cpn60 genes and phenotypic characterization, in order to determine their 60
taxonomic position. Additionally, the whole genome of WBE38
Twas sequenced, since it 61
was selected as the type strain. Based on our results, we propose and describe these 62
strains as a novel species within the genus Campylobacter.
63
64
Over the period between January 2013 and October 2015, a study was conducted to 65
evaluate the prevalence and diversity of campylobacteria in wild bird faecal samples in the 66
city of Valdivia, South of Chile, where a collection of Campylobacter isolates was obtained
67
(L. Collado, paper in preparation). The samples corresponded to bird’s faecal droppings 68
excreted in public urban parks. Although the specific type of bird corresponding to each 69
specimen remains unidentified, at the time of sampling black-faced ibis (Theristicus 70
melanopis), southern lapwing (Vanellus chilensis) and chimango caracara (Milvago 71
chimango) were the main species observed at these sites. Faecal samples were collected 72
using sterile cotton-tipped swabs that were immediately placed into 8 ml Bolton broth 73
(Oxoid). Incubation was performed at 37 °C for 48 h under micro-aerobic conditions 74
(CampyGen, Oxoid). After enrichment, an aliquot (400 µl) was transferred onto the surface 75
of a 0.45 µm membrane filter (Millipore) which was placed on a Petri dish containing Blood 76
Agar Base (Merck) supplemented with 5% defibrinated sheep blood (Quad Five) and it 77
was allowed to filter passively under room conditions for 30 min. After filtration, the filters 78
were removed with sterile forceps and discarded. The inoculated plates were incubated 79
under the aforementioned conditions (Collado et al., 2014). The 17 novel isolates 80
described here were initially identified as Campylobacter lari by the PCR-RFLP described 81
in Marshall et al. (1999) showing the C1 and C1b restriction patterns with the 82
endonucleases DdeI and BsrI, respectively (data not shown). However, recently this 83
species was divided into two subspecies (C. lari subsp. lari and C. lari subsp. concheus) 84
(Debuyne et al., 2009). Additionally, several C. lari-like new species were described, i.e., 85
C. insulaenigrae (Foster et al., 2004), C. peloridis (Debuyne et al., 2009), C. subantarticus 86
(Debuyne et al., 2010a) and C. volucris (Debuyne et al., 2010b), that collectively have 87
been referred to as Campylobacter lari group (Miller et al., 2014b). Since no available PCR 88
method could differentiate these highly related taxa, all isolates we identified as C. lari by 89
PCR-RFLP were further analyzed by a polyphasic taxonomic approach.
90
91
The 17 isolates were first analysed by means of the enterobacterial repetitive intergenic 92
consensus PCR (ERIC-PCR) technique using the Houf et al. (2002) protocol, in order to 93
avoid further analysis of strains with the same genotype. The electrophoretic patterns were 94
analysed using the Jaccard index and a dendrogram was constructed by using the 95
unweighted pair group linkage analysis (UPGMA) method with the Phoretix 1d Pro 96
software (Totallab). As shown in Fig. 1, all the isolates belonged to different ERIC-PCR 97
types and, consequently, were considered different strains. A nearly complete 16S rRNA 98
gene sequence (1350 bp) of five selected strains (WBE38
T, WBE186, WBE206, WBE215 99
and WBE241), were amplified as described by Vandamme et al. (2006). Additionally, the 100
rpoB, atpA and cpn60 genes were also evaluated with the protocols described by Korczak 101
et al. (2006), Miller et al. (2014a) and Hill et al. (2006), respectively. Due to the high 102
number of strains, only the analysis of rpoB gene sequences included all 17 strains. Both 103
DNA strands of the PCR products were directly sequenced with an ABI 3730 XL automatic 104
DNA sequencer (ABI) by a commercial sequencing facility (Macrogen, Seoul, Korea).
105
Alignment of the sequences was performed with the CLUSTAL W program (Thompson et 106
al., 1994). Phylogenetic trees were reconstructed with the MEGA 6 software (Tamura et al., 107
2011), by using the neighbour-joining method (Saitou & Nei, 1987) with Kimura’s two- 108
parameter substitution model (Kimura, 1980) and the stability of the groupings, estimated 109
by bootstrap analysis (500 replications). Similarity values between the 16S rRNA gene 110
sequence of strain WBE38
Tand the type strains of all members of the genus 111
Campylobacter with valid published names were calculated with the EzTaxon-e server 112
(Kim et al., 2012) obtaining a similarity range of 99.5- 90.7%. The 16S rDNA phylogenetic 113
tree (Fig. 2) clearly indicated that the five strains represented a single species within the 114
genus Campylobacter, most closely related to C. subantarticus (99.5% similarity) and C.
115
lari subsp. concheus (99.4% similarity). The taxonomic position of the new strains within
116
the Campylobacter lari group and the demonstration that they represent a novel lineage 117
were confirmed by the rpoB, atpA and cpn60 phylogenetic trees (Supplementary Fig. 1, 2 118
and 3, respectively).
119
For the whole genome sequencing of strain WBE38
T, genomic libraries were prepared with 120
the Nextera® XT DNA Sample Perparation Kit (Illumina, Inc.) and then sequenced using a 121
MiSeq Illumina platform which produced 5,250,728 pair-end reads (2x150 cycles). After 122
initial quality check reads were assembled with SPAdes (Bankevich et al., 2012) and 123
annotated with Prokka (Seemann, 2014), producing 160 contigs that were deposited in the 124
GenBank under the accession number LXSU00000000. The version described in this 125
paper is version LXSU01000000. The average nucleotide identity (ANI) was used as an 126
alternative to DNA-DNA hybridization (DDH) (Konstantinidis & Tiedje, 2005). The ANIb 127
(based on BLAST) values of WBE38
Tcompared to the sequenced type strains of all other 128
species of the C. lari group were calculated using JSpecies v1.2.1 (Richter & Rosselló- 129
Mora, 2009) and were below the 95% species cut-off (Table 1) (Konstantinidis & Tiedje, 130
2005). The genomic G+C content of WBE38
Twas calculated using in-house R scripts and 131
resulted in 29.5%, which is within the range reported for the genus Campylobacter (29- 132
47%) (Debruyne et al., 2008).
133
134
For the physiological and biochemical characterizations of all the new strains, phenotypic 135
testing was performed as described previously (On & Holmes, 1991a, b, 1992; Ursing et 136
al., 1995). Growth of strains was determined on nutrient broth no. 2 (Oxoid) supplemented 137
with 5% defibrinated sheep blood (Quad Five) and 2% agar (Merck). Micro-aerobic growth 138
(using CampyGen, Oxoid) was evaluated at 25°C, 37°C and 42°C for 48 to 72h. Aerobic 139
and anaerobic growths (using AnaeroGen, Oxoid) were evaluated at 37°C for 72h.
140
Catalase activity was evaluated by adding a 3% H
2O
2solution and observing the reaction 141
within 5s. Oxidase activity was determined with Bactident Oxidase strips (Merck). Indoxyl 142
acetate hydrolysis was determined as described by Mills & Gherna (1987). In addition to 143
this, a set of additional phenotypic tests (reduction of nitrates, esterase activity, hydrolysis 144
of hippurate, γ-glutamyl transferase activity, reduction of triphenyl-tetrazolium chloride 145
(TTC), alkaline phosphatase activity, production of H
2S, assimilation of glucose and 146
pyrrolidonyl-, L-arginine- and L-aspartate- arylamidase activities) were evaluated by using 147
the API Campy identification system (bioMérieux) according to the manufacturer’s 148
instructions. All tests were performed at least twice with C. jejuni (DSM 4688
T), C. coli 149
(DSM 4689
T), C. lari (DSM 11375
T), C. subantarticus (LMG 24377
T), C. insulaenigrae 150
(LMG 22716), C. volucris (LMG 24380
T) and Escherichia coli (ATCC 25922) used as 151
controls. The novel isolates were biochemically different from most of the recognized 152
species of the genus Campylobacter because oxidase activity was not detected. This test 153
is positive in all species except C. gracilis and sporadic isolates of C. concisus and C.
154
showae (Vandamme et al., 2005). However, the new isolates could be differentiated from 155
oxidase negative campylobacters by the urease test. Table 2 shows the most important 156
phenotypic characteristics differentiating the novel strains from the other species of the 157
genus Campylobacter. Bacterial cell shape was observed using a scanning electron 158
microscope (Zeiss, LEO 420) following the protocol described by Kawamura et al. (2015).
159
The organism exhibits a curved shape and cells became spherical or coccoid after 72h of 160
incubation (Fig. 3).
161
162
In conclusion, the results from this taxonomic study clearly demonstrate that the isolates 163
recovered from wild birds faecal samples comprise a novel species distinct from other 164
currently known Campylobacter species, based on 16S rRNA, housekeeping genes, whole
165
genome sequence comparison, morphological, physiological and biochemical properties.
166
The name Campylobacter ornithocola sp. nov. is proposed, with WBE38
T(=CECT 9147
T= 167
LMG 29815
T) as the type strain.
168
169
Description of Campylobacter ornithocola sp. nov.
170
Campylobacter ornithocola [or.ni.tho'co.la. Gr. n. ornis, -ithos bird; L. suff. -cola 171
(from L. n. incola) dweller; N.L. n. ornithocola bird dweller].
172
Cells are Gram-negative curved rods, non-encapsulated, non-spore-forming and are 0.3–
173
0.5 µm wide and 1.2–3 µm long. After incubation on Colombia agar (5% sheep blood) in 174
microaerobic atmosphere at 37 °C for 48 h, colonies are glossy, slightly convex, round with 175
smooth margins. Coccoid cells were observed in old cultures. Swarming on solid media 176
was noted. Pigments are not produced. Grows on blood agar at 37 °C and at 42 °C under 177
micro-aerobic culture conditions (does not require atmospheric hydrogen) and at 37°C in 178
anaerobic conditions. No growth was observed at 37 °C on aerobiosis and at 25 °C under 179
micro-aerobic conditions. No haemolysis is seen on blood agar. Catalase- and urease- 180
activity is present but no oxidase. Esterase activity and reduction of Triphenyltetrazolium 181
chloride (TTC) are variable.
182
Pathogenicity is unknown; strains have been recovered from faecal samples of wild birds.
183
The type strain is WBE38
T(=CECT 9147
T= LMG 29815
T) which was isolated from a faecal 184
sample in Valdivia, Chile.
185
186
187
188
Acknowledgements 189
We thank Bernhard Schink (University of Konstanz, Konstanz, Germany) for his help with 190
the specific etymology and nomenclature. We also thank Ricardo Silva (Universidad 191
Austral de Chile) for his help with SEM.
192
193
Funding information 194
This work was supported by the Comisión Nacional de Investigación Científica y 195
Tecnológica (CONICYT–Chile) under the project FONDECYT Nº 11130402.
196
197
Conflicts of interest 198
The authors have no conflict of interest to declare.
199
200
Ethical statement 201
No experiments with humans or animals were carried out.
202
203
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318
Table 1. Average nucleotide identity based on BLAST (ANIb) values (in percentages) for 319
C. ornithocola sp. nov. and the most closely related members of the genus Campylobacter 320
Strains: 1, C. ornithocola sp. nov. WBE38
T; 2, C. lari subsp. concheus LMG 11760; 3, C.
321
lari subsp. lari RM 2100; 4, C. subantarticus LMG 24377
T; 5, C. peloridis LMG 23910
T; 6, 322
C. volucris LMG 24379; 7, C. insulaenigrae NCTC 12927 323
324
Strains 1 2 3 4 5 6 7
1 -
2 90,8 -
3 89,5 92,8 - 4 88,4 89,7 88,8 - 5 86,0 86,2 86,1 85,3 - 6 82,0 82,3 83,1 81,8 82,4 - 7 80,9 81,0 81,2 80,2 81,1 82,9 - 325
326
327
328
329
330
331
332
333
334
335
Table 2. Phenotypic characteristics that differentiate Campylobacter ornithocola sp. nov. from other Campylobacter species.
336
Campylobacter species: 1, C. ornithocola sp. nov (n = 17); 2, C. avium; 3, C. canadensis; 4, C. coli; 5, C. concisus; 6, C. corcagiensis; 7, C. cuniculorum; 8, C. curvus; 9, C. fetus
337
subsp. fetus; 10, C.fetus subsp. testudinum; 11, C. fetus subsp. venerealis; 12, C. geochelonis; 13, C. gracilis; 14, C. helveticus; 15, C. hepaticus; 16, C. hominis; 17, C. hyointestinalis
338
subsp. hyointestinalis; 18, C. hyointestinalis subsp. lawsonii; 19. C. iguanorum; 20, C. insulaenigrae; 21, C. jejuni subsp. doylei; 22, C. jejuni subsp. jejuni; 23, C. lanienae; 24, C. lari
339
subsp. concheus; 25, C. lari subsp. lari; 26, C. mucosalis; 27, C. peloridis; 28, C. rectus; 29, C. showae; 30, C. sputorum; 31, C. subantarticus; 32, C. upsaliensis; 33, C. ureolyticus;
340
34, C. volucris. Data for reference taxa were taken from Foster et al., 2004; Matsuda and Moore, 2004; Debruyne et al., 2008, 2009; 2010a,b; Piccirillo et al., 2016; Van et al., 2016. +,
341 100% strains positive; -, 100% strains negative; (+), 80–94% strains positive; V, 42–66% strains positive; (-), 7–33% strains positive; ND, not determined.
342
Characteristic 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34
Oxidase -* + + + V + + + + + + + - + + + + + + + + + + + + + + + V + + + + +
Catalase + V V + - + + - + + (+) V V - + - + + + + V + + + + - + (-) + V + - V +
Urease + - V - - + - - - ND - - - -† - + -†
Alkaline
phosphatase - - - - V + - V - ND - - - - ND - - (-) - ND - - + - - (+) ND - - - ND - - -
Reduction of
Nitrate V + V + (-) (+) + + + + (+) + (+) + V - + + + + - + + + + (-) ND + + (+) +‡ + + +
2,3,5,
triphenyltetrazolium chloride (TTC)
(-) - ND + - - V V - + - - - - ND - - - ND ND V + ND + + - ND - - - ND V - -
Hydrolysis of
Hippurate - + - - - (-) - - - + - - (+) - - - + + - - - -
Indoxyl acetate - + - + - V + V - - - - V + + - - - + + - ND - - - + V - - + V -
H2S production
(TSI) - - V - - + - (-) - - - + + + - - - - -† - + ND - V + - - - -
α-Haemolysis - - - (-) (-) - + (-) - ND V - - + - - V V - ND + + + ND + - ND + + + + + V ND
Growth at/in/on 25°C
(microaerobic) - - - ND - - + + - + - - - + - - - -
37°C
(microaerobic) + + + + + + + V + + + + - + + + + + + + + + + + + + + - V + + + + +
42ºC
(microaerobic) + + + + (+) + (+) V (+) V - - V + + (-) + + + - - + + + + + + (-) V + + + V +
37°C (anaerobic) + - + - + + - + (-) + V + + - - + - + + - - - + ND - + ND + + + + - V +†
37°C (aerobic) - - - V - - - -
Glycine 1% + - V (+) (+) + - + + + (-) + + V + + V + + (-) + - (+) + V + + V + (+) + + -
H2 requirement - V - - + - - + - - - - + - - + V V - ND - - - ND - + ND + + - - - - ND
DNA G+C (mol %)
29.5 35 ND 31 37-
41 31.9 32.4 45- 46
33-
35 ND 33-
34 33.6 44-
46 34 27.9 32.5 35-
36 31-
33 36 ND 31 30-
31 36 30 29-
30 36-
38 29 45-
46 44- 46
29-
33 30 32-
36 28-
30 29
* Oxidase negative test allows differentiating the new species from Urease-Positive Thermophilic Campylobacter (UPTC) strains.
343
† This test was determined for the corresponding type strain, in this study
344
‡ We obtained a different result from that published by Debruyne et al. (2010b), when we evaluated the nitrate test with the API Campy (nitrate reduction negative for strain LMG
345
24377
T).
346 347
348
349
350
351
352
353
354
Figures legends 355
Fig. 1. Cluster analysis based on the patterns obtained by ERIC-PCR from the 356
Campylobacter ornithocola sp. nov. strains.
357
Fig. 2. Neighbour-joining tree based on 16S rRNA sequences showing the phylogentic 358
position of Campylobacter ornithocola sp. nov. within the genus Campylobacter. Bootstrap 359
values >70%, generated from 500 replicates, are shown at the nodes. The scale bar 360
represents substitutions per site.
361
Fig. 3. Images of cells of strain WBE38
Tas observed with scanning electron microscopy.
362
The organism exhibits a curved shape while spherical or coccoid forms can also be 363
observed. Bars, 3 µm.
364
Figure 1 Click here to download Figure Fig. 1 (ERIC-PCR).pptx
C. ornithocolaWBE 215 (KX467977) C. ornithocolaWBE 241 (KX467978) C. ornithocolaWBE 206 (KX467976) C. ornithocolaWBE 186 (KX467975) C. ornithocolaWBE 38T (KX467974)
C. lari UPTC NCTC 11845 (CP007775)
C. lari subsp. concheusLMG 21009T (AM922330) C. subantarcticusLMG 24377T (AM933371)
C. volucris LMG 24380T(FM883694) Campylobacter sp. RM16704 (CP007769)
C. lari subsp. lariATCC 35221T(AY621114) C. peloridis LMG 23910T(AM922331) C. hepaticusNCTC 13823T(KU886019) C. insulaenigrae NCTC 12927T(AJ620504) C. jejuni subsp. doyleiLMG 8843T(DQ174144) C. jejuni subsp. jejuniNCTC 11351T(AF372091)
C. coli LMG 6440T(AF372092)
C. cuniculorumLMG 24588T(DQ400345) C. helveticusATCC 51209T(U03022)
C. aviumLMG 24591T(EU623473) C. upsaliensisCCUG 14913T(DQ174157)
C. canadensisLMG 24001T(EF621894) C. hyointestinalissubsp. lawsoniiLMG 14432T(AF097685)
C. lanienaeNCTC 13004T(AF043425) C. iguaniorumLMG 28143T(KF425533)
C. hyointestinalissubsp. hyointestinalisNCTC 11608T(AF097689) C. fetussubsp. testudinumLMG 27499T(CP006833)
C. fetussubsp. fetusATCC 27374T(DQ174127) C. fetussubsp. venerealisNCTC 10354T(DQ174131)
C. corcagiensisCIT045T(KF745861) C. ureolyticusATCC 33387T(L04321)
C. mucosalisATCC 43264T(DQ174173) C. concisusATCC 33237T(L04322)
C. sputorumbiovarparaureolyticusLMG 17590 (AF022768) C. sputorumbiovarsputorumLMG 7795 (X67775) C. sputorumbiovarfecalisLMG 6617 (AF550637) C. curvusATCC 35224T(DQ174165)
C. hominisNCTC 13146T(AJ251584) C. gracilisATCC 33236T(DQ174168)
C. rectusATCC 33238T(L04317)
C. showaeCCUG 30254T(DQ174155)
100 99
99
100 100 98
91
93 89
94
92
79 94 88
100 92
90 80 98
0.01
Figure 2 Click here to download Figure Fig. 2 (16S rRNA).pptx
Figure 3 Click here to download Figure Fig. 3 (SEM).pptx
Supplementary data on-line International Journal of Systematic and Evolutionary Microbiology
Campylobacter ornithocola sp. nov., a new member of the Campylobacter lari group isolated from wild bird faecal samples
Alberto Cáceres
1, Ivo Muñoz
1, Gregorio Iraola
2, Florencia Díaz-Viraqué
3and Luis Collado
11
Institute of Biochemistry and Microbiology, Faculty of Sciences, Universidad Austral de Chile, Valdivia, Chile.
2Bioinformatics Unit, Institut Pasteur Montevideo, Montevideo, Uruguay.
3Molecular Biology Unit, Institut Pasteur Montevideo, Montevideo, Uruguay.
Corresponding Author:
Luis Collado González
Phone: 56-63-2293848
Fax: 56-63-2215295
E-mail: luiscollado@uach.cl
Supplementary Material Files Click here to download Supplementary Material Files
supplementary data.pdf
C. ornithocola WBE 249 (KX468004) C. ornithocola WBE 258 (KX468005) C. ornithocola WBE 241 (KX468003) C. ornithocola WBE 225 (KX468002) C. ornithocola WBE 215 (KX468001) C. ornithocola WBE 212 (KX468000) C. ornithocola WBE 210 (KX467999) C. ornithocola WBE 206 (KX467998) C. ornithocola WBE 201 (KX467997) C. ornithocola WBE 196 (KX467996) C. ornithocola WBE 195 (KX467995) C. ornithocola WBE 38T (KX467989) C. ornithocola WBE 186 (KX467990) C. ornithocola WBE 187 (KX467991) C. ornithocola WBE 188 (KX467992) C. ornithocola WBE 189 (KX467993) C. ornithocola WBE 193 (KX467994) C. lari UPTC NCTC 11845 (CP007775)
C. subantarticus LMG 24377T (KX901855)
C. lari subsp. concheusLMG 121009T (KX901854) C. lari subsp. lariNCTC 11352T(DQ174202) Campylobactersp. RM16704 (CP007769)
C. peloridis LMG 23910T(CP007766) C. volucris LMG 24380T (KX901853) 93
75 85
99
Supplementary Fig. 1. Neighbour-joining tree based on rpoB sequences showing the phylogentic position of Campylobacter ornithocola sp. nov. within Campylobacter lari group. Bootstrap values >70%, generated from 500 replicates, are shown at the nodes.
The scale bar represents substitutions per site.
C. ornithocolaWBE 38T(KX467979)
C. ornithocolaWBE 186 (KX467980)
C. ornithocolaWBE 241 (KX467983)
C. ornithocolaWBE 206 (KX467981)
C. ornithocolaWBE 215 (KX467982)
Campylobacter lariUPTC NCTC 11845 (CP007775)
C. subantarcticusLMG 24377T(KF855300)
C. larisubsp. concheusLMG 11760 (KF855291)
C. larisubsp. lariRM 2100 (KF855290)
C. peloridisLMG 23910T(KF855294)
Campylobactersp. RM16704 (CP007769)
C. insulaenigraeNCTC 12927T (KF855286)
C. volucrisLMG 24379 (KF855304)
C. fetussubsp. fetus82-40 (CP000487) 99
89 99 91
96
77
