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Gallibacterium
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Genetic diversity of Gallibacterium isolates from
California turkeys
a b
A. M. Bojesen & H. L. Shivaprasad
a
Department of Veterinary Pathobiology, Faculty of Life Science, University of
Copenhagen, Stigb⊘⊘jlen 4, DK-1870, Frederiksberg C, Denmark
b
California Animal Health and Food Safety Laboratory System, Fresno Branch, School of
Veterinary Medicine, University of California, Davis, California, USA
Available online: 11 May 2007
To cite this article: A. M. Bojesen & H. L. Shivaprasad (2007): Genetic diversity of Gallibacterium isolates from California
turkeys, Avian Pathology, 36:3, 227-230
To link to this article: http://dx.doi.org/10.1080/03079450701332352
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2. Avian Pathology (June 2007) 36(3), 227 Á 230
Genetic diversity of Gallibacterium isolates from California
turkeys
A. M. Bojesen1* and H. L. Shivaprasad2
1
Department of Veterinary Pathobiology, Faculty of Life Science, University of Copenhagen, Stigbøjlen 4, DK-1870
Frederiksberg C, Denmark, and 2California Animal Health and Food Safety Laboratory System, Fresno Branch, School of
Veterinary Medicine, University of California, Davis, California, USA
The aim of the present study was to investigate the genetic diversity of Gallibacterium isolates recovered
from lesions in turkeys. Gallibacterium has been isolated from various bird species including turkeys, but no
large investigations have yet been made to characterize isolates from turkeys genetically. We therefore
genotyped 53 Gallibacterium isolates obtained from turkeys between 1998 and 2004. Fifty isolates originated
from 29 different flocks in California and the remaining three came from three German turkey flocks. All
were recovered from birds with lesions, mainly in the upper respiratory tract. Five chicken isolates from
California and five Gallibacterium reference strains were also included. Amplified fragment length
Downloaded by [190.234.159.185] at 10:09 12 September 2011
polymorphism analysis demonstrated substantial genetic diversity among the Gallibacterium isolates.
However, we also demonstrated that some Gallibacterium clones were present in consecutive rotations at the
same farm during the entire 6-year observation period and were present in different flocks from
different farms. Similarly, the same clone was identified from two of the three German flocks. Further in-
vestigation of the spread of Gallibacterium between turkey flocks, including infections acquired from
chickens or wild birds, should be carried out.
Introduction
The taxonomy of organisms previously reported as analysis (Zellner et al ., 2004). However, little is known
Actinobacillus salpingitidis, avian Pasteurella haemoly- about the epidemiological aspects of Gallibacterium
tica or Pasteurella anatis has recently been re-investi- infections in turkeys at farm, flock or at host level.
gated, leading to formation of a new genus, Genotypic methods involving characterization of the
Gallibacterium (Christensen et al ., 2003). The impor- whole genome such as pulsed-field gel electrophoresis
tance of Gallibacterium as a pathogen is not fully and amplified fragment length polymorphism (AFLP)
understood although a number of reports have indicated have been used successfully to clarify epidemiological
a pathogenic potential. Isolates have been recovered aspects of Gallibacterium infections in chicken flocks but
from mixed infections and in pure culture from a range isolates from turkeys have never been investigated on a
of pathological lesions in poultry, including septicaemia, large scale (Bojesen et al ., 2003b; Christensen et al .,
oophoritis, follicle degeneration, salpingitis, peritonitis, 2003). AFLP and pulsed-field gel electrophoresis repre-
enteritis and respiratory tract lesions (Kjos-Hanssen, sent highly reproducible and discriminatory genotyping
1950; Kohlert, 1968; Mraz et al ., 1976; Gerlach, 1977;
´ methods allowing identification of clonal lineages in
Matthes & Hanschke, 1977; Mushin et al ., 1980; bacterial populations (Vos et al ., 1995; Spratt & Maiden,
Bisgaard & Dam, 1981; Shaw et al ., 1990, Mirle et al ., 1999).
1991; Bojesen et al ., 2003a; Jordan et al ., 2005). Consequently, the aim of the present study was to
Bacteria belonging to the genus Gallibacterium seem characterize a collection of G. anatis bv. haemolytica
to have a wide host spectrum based on isolations from isolates from turkeys by AFLP and to evaluate popula-
domestic birds as well as a range of non-domestic birds tion dynamics over time and within natural populations.
including chickens, turkeys, geese, ducks, pheasants,
partridges, cattle egrets and others (Mushin et al .,
1980; Bisgaard, 1993). Most studies of this organism Materials and Methods
have been in chickens, and isolates obtained from other
Bacterial strains. Fifty-three G. anatis bv. haemolytica isolates from
hosts have not been characterized to any great extent.
turkeys were characterized in this study. The California isolates were
A recent study reporting the occurrence of Gallibacter- isolated between 1998 and 2004 from a total of 29 flocks owned by five
ium anatis biovar haemolytica in California diagnostic different operations. An additional five isolates from chickens, also
avian specimens during 1994 to 2003 stated that from California, were included for comparison. The chicken isolates
G. anatis bv. haemolytica was isolated from a total of originated from five epidemiologically unrelated flocks and were
152 out of 3609 meat turkeys subjected to postmortem isolated in 2004. All isolates were retrieved from accessions in the
*To whom correspondence should be addressed. Tel: '45 35282781. Fax: '45 35282757. E-mail: miki@life.ku.dk
Received 28 November 2006
ISSN 0307-9457 (print)/ISSN 1465-3338 (online)/07/30227-04 # 2007 Houghton Trust Ltd
DOI: 10.1080/03079450701332352
3. 228 A. M. Bojesen and H. L. Shivaprasad
California Animal Health and Food Safety Laboratory System. The genotypic results from the current investigation strongly
remaining three isolates were obtained from three German turkey indicate that all of the included isolates have been
flocks. Five Gallibacterium reference strains (CCM5974, CCM5975,
correctly identified as Gallibacterium .
CCM5976, F149T and 12656-12), including the type strain, were also
included.
The AFLP procedure was repeated only for those
All the field strains were isolated from lesions, mainly in the upper isolates (approximately 5%) that generated fingerprints
respiratory tract. Other agents were also identified in most of the cases, that did not allow proper analysis in the first run. Repeat
the most prevalent being Ornitobacterium rhinotracheale (six cases), reactions were deemed unnecessary for other isolates as
Bordetella avium (six cases), Escherichia coli (five cases), Pasteurella our earlier study using the same AFLP protocol had a
multocida (one case), Mycoplasma spp. (one case) or a mixed bacterial reproducibility of 97.5%, indicating that it was very
population mainly composed of two or more of the above agents robust (Bojesen et al ., 2003b).
(24 cases) (Zellner et al , 2004). Swabs were plated on blood agar base
A total of 41 individual Gallibacterium clones were
supplemented with 5% bovine blood and the plates incubated overnight
at 378C in plastic bags. Circular smooth, shiny greyish raised colonies
identified among the 63 isolates using the 90% Dice
with a diameter of 1.0 to 2.0 mm after 24 h at 378C and appearing similarity cut-off point. This cut-off limit has been used
strongly b-haemolytic, opaque, with a butyrous consistency were in previous studies employing AFLP to assess the
subcultured on blood agar plates to obtain pure cultures. Frozen stock genetic diversity of Gallibacterium and other bacteria
cultures were subsequently made from cultures in heart infusion broth (Janssen et al ., 1997; Savelkoul et al ., 1999; Bojesen
(Difco, Detroit, Michigan, USA) incubated overnight at 378C. Seven et al ., 2003b). Of the 41 clones, 33 appeared as single
hundred microlitres of heart infusion broth culture were mixed with isolates whereas the remaining eight were part of clonal
300 ml sterile 50% (v/v) glycerol and stored at (808C.
complexes as defined as groups of identical (SD ]90%)
bacterial isolates (Figure 1). Three of the complexes
Bacterial identification. All isolates were subjected to phenotypic contained from three to nine identical isolates. Interest-
characterization in accordance with Bisgaard (1982) and Christensen ingly, the same clone was isolated at different time points
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et al . (2003), and genotypic identification by the recently described
from operation A over the entire observation period
Gallibacterium -specific polymerase chain reaction (PCR) (Bojesen
et al ., 2007). This PCR is based on primers targeting the 16S and 23S
from 1998 to 2004 (Figure 1). A similar picture was seen
rRNA genes, generating two or three amplicons depending on the with the German turkey isolates where two flocks were
number of different ribosomal operons present. An internal amplifica- infected with the same clone.
tion control based on the rpoB gene is also a part of the PCR protocol. One of the Californian clones was isolated in one case
from turkeys originating from six different flocks owned
Genotyping by AFLP. All isolates were characterized as described by by two different operations (Figure 1). Strict biosecurity,
Bojesen et al . (2003b). Briefly, chromosomal DNA was extracted from including hygiene barriers and visitor clearing, was
cell pellets of overnight cultures in heart infusion broth by the use of practiced in all the operations included in our investiga-
Easy-DNA kitTM (Invitrogen, Carlsbad, California, USA). The AFLP tion although none of them operated on an ‘all in all out’
reactions were based on the non-selective Bgl II primer (FAM-5?-
basis. For example, there were eight to 15 houses on each
GAGTACACTGTCGATCT-3?) and the non-selective Bsp DI primer
farm and the individual houses contained birds of
(5?-GTGTACTCTAGTCCGAT-3?) and were used to amplify the
fragments subsequent to restriction digestion and ligation to their different ages. Each farm was serviced by two or three
corresponding adaptors. Amplification products were detected on an workers, who did not visit flocks on any other farms.
ABI 377 automated sequencer (PE Biosystems). Each lane included However, there was traffic between farms by trucks
internal-lane size standard labelled with ROX dye (Applied Biosystems), carrying feed, live and dead birds. In addition, all farms
and GeneScan 3.1 fragment analysis software (Applied Biosystems) was were located in areas that were densely populated with
used for fragment size determination and pattern analysis. AFLP commercial poultry including chickens and turkeys.
profiles comprising fragments in the size range 50 to 500 base pairs were Gallibacterium organisms are not expected to survive
considered for numerical analysis with the program GelCompar II
well in poultry houses between rotations due to the
(Applied Maths, Kortrijk, Belgium). Normalized AFLP fingerprints
were compared using the Dice similarity coefficient with a 0.5% cleaning and disinfection procedures and their poor
optimization and 0.1% position tolerance, and clustering analysis was viability outside the host (Bisgaard, 1993), but the spread
performed by the unweighted pair group method with arithmetic of these organisms and other pathogens between flocks
averages. and farms, for example by the wind, remains a possibi-
lity. There are other examples of bacterial clones, as in
the case of P. multocida , that have been transmitted
Results and Discussion between poultry from different production systems and
All included turkey isolates were b-haemolytic and between farmed and wild birds (Christensen et al ., 1998;
classified with Gallibacterium based on the phenotypic Pedersen et al ., 2003).
characters tested. Similarly, Gallibacterium -specific In addition to isolation from various types of poultry,
amplicons at the expected sizes of 780 and 1060 base Gallibacterium organisms have been isolated from
pairs were generated by PCR in all but one isolate different bird species kept in captivity and from wild
(F0101711; data not shown). In addition, an intermedi- birds (Bisgaard, 1993). However, virtually nothing is
ate size amplicon of Â900 base pairs was present in all known about possible interchange of organisms between
isolates. Only the small and intermediate-sized ampli- these different bird populations. Although the current
cons were present in F0101711. An amplicon of inter- study was focused on Gallibacterium isolated from
mediate size indicating the existence of a third size turkeys, a small number of isolates from chickens, also
ribosomal operon has been described previously by from California, were characterized by AFLP. Interest-
Christensen et al . (2003); however, based on the previous ingly, the California chicken isolates that were included
and present results this operon does not seem to occur appeared to be related to the turkey isolates, and in one
very frequently in Gallibacterium (Bojesen et al ., 2007). case even grouped together with a turkey clone, indicat-
The internal amplification control based on the rpoB ing that there could be a link between the different
gene was positive in all samples. The phenotypic and production systems (Figure 1).
4. Diversity of Gallibacterium isolates from turkeys 229
SD(%)
100
30 Strain no. Host Origin Operation Farm Age (wks)
35
40
45
50
55
60
65
70
75
80
85
90
95
CCM5975 Chicken
CCM5976 Chicken
CCM5974 Chicken
F0303525 Turkey Sinus A PEN 3
F9804343 Turkey Air sac A SIX 8
F149T Duck
F0401588 Chicken
F0203227 Turkey Trachea A FRO 14
F0203779 Turkey Trachea A HAN 5
F0304008 Turkey Trachea A HUN 5
F0304605 Turkey ND A LAK 9
F0002299 Turkey Air sac A CRCK 2
F0102568 Turkey Trachea D MCM 16
F0103100 Turkey Sinus A HUN 5
F0203942 Turkey Trachea A SIX 7
F0202732 Turkey Sinus A S&W 7
F0000732 Turkey Trachea D MCM 12
F0304244 Turkey Lung A HUN 7
F0400243 (As2) Turkey Air sac A HUN 11
F0304695 Turkey Air sac A LAK 7
F0400748 Chicken
F0403502 Turkey ND A HUN 6
1743 Turkey Trachea G (Germany) UK
1797 Turkey Joint H (Germany) UK
1756 Turkey Trachea I (Germany) UK
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F9903469 Turkey Sinus A KER 12
F0103596 Turkey Trachea A KER 4
F9903350 Turkey Trachea A KBO 10
F0004359 Turkey Sinus A PLA 10
F9800718 Turkey Sinus A PAR 7
F9903108 Turkey Air sac A SIX 7
F9902075 Turkey Trachea A SIX 17
F0001986 Turkey Lung A SIX 12
F0103287 Turkey Air sac A HUN 7
F0101794 Turkey Trachea A SIX 9
F0003036 Turkey Lung A SIX 8
F0002646 Turkey Trachea A SIX 3
F0002647 Turkey ND A SIX 11
F0002767 Turkey Air sac A MAR 8
F0400496 Turkey Lung A MAR 11
F9800812 Turkey Air sac A MAR 15
F0400193 Chicken
F0401624 Chicken
F0003406 Turkey Liver A TWI 7
F0203184 Turkey Heart sac A SMT 9
F9804556 Turkey Air sac A TWI 4
F0400159 Chicken
F0400243 (Tr1) Turkey Trachea A HUN 11
F0104108 Turkey Trachea E DIN 8
F0103931 Turkey Trachea E DIN 7
F0304601 Turkey Sinus A RSM 17
F0102911 Turkey Trachea A TCR 7
F0302082 Turkey ND A ARV 14
F0303555 Turkey ND A FRY 18
F0402657 Turkey ND B NOR 7
F0403791 Turkey ND A SIX 6
F0404303 Turkey ND A SIX 10
F0301778 Turkey Joint A CLA 36
F0101710 Turkey Joint E DIN 22
12656-12 Liver Chicken
F0304489 Turkey Trachea A SIX 13
F9803422 Turkey Trachea A SIX 13
F0101711 Turkey Joint C CHR 20
Figure 1. Dendrogram showing AFLP similarities (Dice coefficient) between the Gallibacterium isolates obtained from turkeys. Vertical
dotted line at 90% similarity denotes the cut-off value for a clone. Shaded boxes indicate groups of isolates belonging to the same clonal
complex. The year of isolation is indicated in the first two digits of the strains number (i.e. 9801998).
The five Gallibacterium reference strains were only in each flock (Bojesen et al ., 2003b). However, isolates
distantly related to the California isolates, but this was from only two flocks were characterized, and the
expected due to the large temporal and geographical characterization of isolates from more flocks is probably
separation between them. A previous study assessing the necessary to substantiate this observation.
genetic diversity among Gallibacterium isolates from In conclusion, we have demonstrated a high
chickens found that typically only one clone was present genetic diversity among Gallibacterium isolates from
5. 230 A. M. Bojesen and H. L. Shivaprasad
Californian turkeys. In addition, we have shown that Gerlach, H. (1977). Significance of Pasteurella haemolytica in poultry.
some Gallibacterium clones were present in consecutive Der Praktische Tierarzt , 58 , 324 Á328.
Kjos-Hanssen, B. (1950). Oviduct peritonitis in hens due to pathogenic
flocks on the same farm over 6 years and in different
"cloacal bacteria". Nordisk Veterinaer Medicin , 2 , 523 Á531.
flocks at different farms. Further studies are required to ¨
Kohlert, R. (1968). Untersuchrungen zur Atiologie der Eileiterent-
provide more insight into the clonal spread and persis- zundung beim Huhn. Monatshefte fur Veterinarmedizin , 23 , 392 Á
¨ ¨ ¨
tence of Gallibacterium . 395.
Janssen, P., Maquelin, K., Coopman, R., Tjernberg, I., Bouvet, P.,
Kersters, K. & Dijkshoorn, L. (1997). Discrimination of Acineto-
Acknowledgement bacter genomic species by AFLP fingerprinting. International Journal
of Systematic Bacteriology, 47 , 1179 Á1187.
Vi Phuong Thi Nguyen is thanked for excellent technical Jordan, F.T.W., Williams, N.J., Wattret, A. & Jones, T. (2005).
assistance. Observations on salpingitis, peritonitis and salpingoperitonitis in a
layer breeder flock. The Veterinary Record , 157 , 573 Á577.
Matthes, S. & Hanschke, J. (1977). Experimentelle Untersuchrungen zur
References ¨
Ubertragung von Bakterien u ¨ber das Huhnerei. Berliner und Munch-
¨ ¨
ener Tierartzliche Wochenschrift , 90 , 200 Á203.
¨
Bisgaard, M. (1982). Isolation and characterization of some previously Mirle, C., Schongarten, M., Meinhart, M. & Olm, U. (1991). Studies
unreported taxa from poultry with phenotypical characters related to into the incidence of Pasteurella haemolytica infections and their
Actinobacillus - and Pasteurella species. Acta Pathologica Microbio- relevance to hens, with particular reference to diseases of the egg-
logica et Immunologica Scandinavica [B] , 90 , 59 Á67. laying apparatus. Monatshefte fur veterinarmedizin , 45 , 545 Á549.
¨ ¨
Bisgaard, M. (1993). Ecology & significance of Pasteurellaceae in Mraz, O., Vladık, P. & Bohacek, J. (1976). Actinobacilli in domestic
´ ´ ´
animals. Zentralblat fur Bakteriologie, 279 , 7 Á26.
¨ fowl. Zentralblat fur Bakteriologie und Hygieine Orig. A , A236 , 294 Á
¨
Bisgaard, M. & Dam, A. (1981). Salpingitis in poultry. II. Prevalence, 307.
bacteriology, and possible pathogenesis in egg-laying chickens. Mushin, R., Weisman, Y. & Singer, N. (1980). Pasteurella haemolytica
Downloaded by [190.234.159.185] at 10:09 12 September 2011
Nordisk Veterinaer Medicin , 33 , 81 Á89. found in respiratory tract of fowl. Avian Diseases, 24 , 162 Á168.
Bojesen, A.M., Nielsen, S.S. & Bisgaard, M. (2003a). Prevalence and Pedersen, K., Dietz, H.H., Jørgensen, J.C., Christensen, T.K., Bregne-
transmission of haemolytic Gallibacterium species in chicken produc- balle, T. & Andersen, T.H. (2003). Pasteurella multocida from
tion systems with different biosecurity levels. Avian Pathology, 32 , outbreaks of avian cholera in wild and captive birds in Denmark.
503 Á510. Journal of Wildlife Diseases, 39 , 808 Á816.
Bojesen, A.M., Torpdahl, M., Christensen, H., Olsen, J.E. & Bisgaard, Savelkoul, P.H.M., Aarts, H.J.M., de Haas, J., Dijkshoorn, L., Duim,
M. (2003b). Genetic diversity of Gallibacterium anatis in chicken B., Otsen, M., Rademaker, J.L.W., Schouls, L. & Lenstra, J.A. (1999).
flocks representing different production systems. Journal of Clinical Amplified-fragment length polymorphism analysis: the state of an
Microbiology, 41 , 2737 Á2740. art. Journal of Clinical Microbiology, 37 , 3083 Á3091.
Bojesen, A.M., Vazquez, M.E., Robles, F., Gonzalez, C., Soriano, E.V., Shaw, D.P., Cook, D.B., Maheswaran, K., Lindeman, C.J. & Halvorson,
Olsen, J.E. & Christensen, H. (2007). Specific identification of D.A. (1990). Pasteurella haemolytica as a co-pathogen in pullets and
Gallibacterium by a PCR using primers targeting the 16S rRNA laying hens. Avian Diseases , 34 , 1005 Á1008.
and 23S rRNA genes. Veterinary Microbiology. DOI: 10.1016/j. Spratt, B.G. & Maiden, M.C. (1999). Bacterial population genetics,
vetmic.2007.02.013 evolution and epidemiology. Philosophical Transactions by the Royal
Christensen, J.P., Dietz, H.H. & Bisgaard, M. (1998). Phenotypic and Society of London, Biological Sciences, 354 , 701 Á710.
genotypic characters of isolates of Pasteurella multocida obtained Vos, P., Hogers, R., Bleeker, M., Reijans, M., van de Lee, T., Hornes,
from backyard poultry and from two outbreaks of avian cholera in M., Frijters, A., Pot, J., Peleman, J., Kuiper, M. & Zabeau, M. (1995).
avifauna in Denmark. Avian Pathology, 27 , 373 Á381. AFLP: a new technique for DNA fingerprinting. Nucleic Acids
Christensen, H., Bisgaard, M., Bojesen, A.M., Mutters, R. & Olsen, J.E. Research , 23 , 4407 Á4414.
(2003). Genetic relationships among strains of biovars of avian Zellner, D.E., Charlton, B.R., Cooper G. & Bickford, A.A. (2004).
isolates classified as ‘‘Pasteurella haemolytica ’’, ‘‘Actinobacillus sal- Prevalence of Gallibacterium species in California poultry over 10
pingitidis ’’ or Pasteurella anatis with proposal of Gallibacterium years (1994 Á2003) from diagnostic specimens. In Proceedings of the
anatis gen. nov., sp. nov. International Journal of Systematic and Annual Conference of the American Association of Avian Pathologists
Evolutionary Microbiology, 53 , 275 Á287. (p. 64). Philadelphia, PA, USA.
6. Avian Pathology (June 2007) 36(3), 1 Á2
Non-English Abstracts
Genetic diversity of Gallibacterium isolates from California
turkeys
A. M. Bojesen1* and H. L. Shivaprasad2
1
Department of Veterinary Pathobiology, Faculty of Life Science, University of Copenhagen, Stigbøjlen 4, DK-1870
Frederiksberg C, Denmark, and 2California Animal Health and Food Safety Laboratory System, Fresno Branch, School of
Veterinary Medicine, University of California, Davis, California, USA
Diversite genetique des souches de Gallibacterium isolees de dindes en Californie
´ ´ ´ ´
Le but de la presente etude a ete d’investiguer la diversite genetique des souches de Gallibacterium isolees de
´ ´ ´ ´ ´ ´ ´ ´
lesions chez des dindes. Gallibacterium a ete isole de differentes especes d’oiseaux incluant les dindes, mais
´ ´ ´ ´ ´ `
jusqu’a present il n’y a pas eu d’investigations importantes pour caracteriser genetiquement les souches
` ´ ´ ´ ´
isolees chez les dindes. Nous avons donc genotype 53 souches de Gallibacterium isolees de dindes entre les
´ ´ ´ ´
annees 1998 et 2004. Cinquante isolats etaient originaires de 29 troupeaux differents en Californie et les trois
´ ´ ´
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autres venaient de troupeaux de dindes en Allemagne. Tous les isolats obtenus a partir d’oiseaux presentant
` ´
des lesions, principalement au niveau de l’appareil respiratoire superieur. Cinq souches isolees de poulets en
´ ´ ´
Californie et cinq souches de reference de Gallibacterium ont ete incluses dans l’etude. L’analyse du
´ ´ ´ ´ ´
polymorphisme de longueur des fragments geniques amplifies a montre une diversite genetique substantielle
´ ´ ´ ´ ´ ´
parmi les isolats de Gallibacterium. Cependant, nous avons egalement demontre que quelques clones de
´ ´ ´
Gallibacterium etaient presents dans les bandes successives au niveau de la meme ferme durant la periode des
´ ´ ˆ ´
six annees d’observation et etaient presents dans des troupeaux differents d’elevages differents. De meme, le
´ ´ ´ ´ ´ ´ ˆ
meme clone a ete identifie a partir de deux des trois troupeaux allemands. Des investigations supplementaires
ˆ ´ ´ ´` ´
sur la dissemination de Gallibacterium entre les troupeaux de dindes, incluant les infections acquises a partir
´ `
des poulets ou des oiseaux sauvages doivent etre entreprises.
ˆ
Genetische Diversitat bei Gallibacterium-Isolaten aus kalifornischen Puten
¨
Das Ziel der vorliegenden Studie war, die genetische Diversitat von Gallibacterium-Isolaten aus
¨
pathologisch-anatomischen Veranderungen bei Puten zu untersuchen. Gallibacterium ist aus verschiedenen
¨
Vogelspezies einschließlich Puten isoliert worden, aber bislang ist keine großere Untersuchung zur
¨
genetischen Charakterisierung der Putenisolate durchgefuhrt worden. Aus diesem Grund haben wir 53
¨
Gallibacterium-Isolate von Puten aus den Jahren 1998 bis 2004 genotypisiert. 50 Isolate stammten aus 29
verschiedenen Herden in Kalifornien und die ubrigen drei kamen aus drei Putenherden in Deutschland. Alle
¨
Isolate wurden in Puten mit pathologisch-anatomischen Veranderungen hauptsachlich im oberen
¨ ¨
Respirationstrakt nachgewiesen. In die Untersuchungen wurden funf Huhnerisolate aus Kalifornien und
¨ ¨
funf Gallibacterium-Referenzstamme miteinbezogen. Die Langenpolymorphismus-Analyse der amplifizier-
¨ ¨ ¨
ten Fragmente ließ eine beachtliche genetische Diversitat zwischen den Gallibacterium-Isolaten erkennen.
¨
Wir konnten jedoch auch zeigen, dass einige Gallibacterium-Klone wahrend der sechsjahrigen Beobach-
¨ ¨
tungsperiode in aufeinanderfolgenden Durchgangen auf derselben Farm, aber auch bei verschiedenen
¨
Herden auf unterschiedlichen Farmen vorkamen. Ebenso wurde der gleiche Klon auf zwei der drei
¨
deutschen Farmen nachgewiesen. Weitere Untersuchungen zur Ubertragung von Gallibacterium zwischen
Putenfarmen einschließlich der von Huhnern und Wildvo
¨ ¨gel erworbenen Infektionen sollten durchgefuhrt
¨
werden.
Diversidad genetica de ailados de Gallibacterium de pavos de California
´
El objetivo de este estudio fue investigar la diversidad genetica de aislados de Gallibacterium obtenidos de
´
lesiones en pavos. Se ha aislado Gallibacterium de distintas especies incluyendo pavos, pero aun no se han
´
realizado estudios mas amplios para caracterizar geneticamente aislados de pavos. Por lo tanto, se
´ ´
genotiparon 53 aislados de Gallibacterium obtenidos de pavos entre 1998 y 2004. Cincuenta aislados
procedıan de 29 lotes distintos de California y los otros 3 procedıan de lotes de pavos de Alemania. Todos se
´ ´
*To whom correspondence should be addressed. Tel: '45 35282781. Fax: '45 35282757. E-mail: miki@life.ku.dk
Received 28 November 2006
ISSN 0307-9457 (print)/ISSN 1465-3338 (online)//30001-02 # 2007 Houghton Trust Ltd
DOI: 10.1080/03079450701332352
7. 2 A. M. Bojesen and H. L. Shivaprasad
obtuvieron de animales con lesiones, principalmente en el tracto respiratorio superior. Tambien se ´
incluyeron cinco aislados de pollos de California y cinco cepas de referencia de Gallibacterium. El analisis
´
del polimorfismo de la longitud de los fragmentos de restriccion demostro una diversidad genetica
´ ´ ´
importante entre los aislados de Gallibacterium. Sin embargo, tambien demostramos que algunos clones de
´
Gallibacterium estaban presentes en rotaciones consecutivas en la misma granja durante todo el perıodo de
´
observacion de seis anos y en lotes distintos de granjas distintas. De modo similar, se identifico el mismo
´ ˜ ´
clon en dos de los tres lotes de Alemania. Deberıan llevarse a cabo mas estudios de la transmision de
´ ´ ´
Gallibacterium entre lotes de pavos, incluyendo infecciones causadas por pollos o aves salvajes.
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